THE CABINET of MINISTERS of UKRAINE

NATIONAL UNIVERSITY of LIFE and ENVIROMENTAL SCIENCES of

UKRAINE

Biotechnology facultyEcobiotechnology and biodiversity department

"APPROVED"

The dean of Biotechnology faculty

_____________Y. V. Kolomietc

“___” _______________ 2013

SCIENTIFIC-METHODOLOGY COMPLEXDiscipline

"Industrial biotechnology"

For grounding of specialists in direction 6.051401- "Biotechnology" in agrarian higher educational institutions

of III – IV levels of accreditation

KYIV - 2013

Form № Н - 3.04

THE CABINET of MINISTERS of UKRAINE

NATIONAL UNIVERSITY of LIFE and ENVIROMENTAL SCIENCES of

UKRAINE

Biotechnology facultyEcobiotechnology and biodiversity department

"APPROVED"

The dean of Biotechnology faculty

_____________Y. V. Kolomietc

“___” _______________ 2013

Working program of discipline«Industrial biotechnology»

For grounding of specialists in direction 6.051401- "Biotechnology"in agrarian higher educational institutions

of III – IV levels of accreditation

KYIV - 2013

Working educational program was completed by Associate ProfessorEcobiotechnology and biodiversity department V. Borodai.Working educational program was discussed at the Ecobiotechnology andbiodiversity departmentReport № _________from _______________---

Head of department, Associate Professor O. L. Klyachenko

Decreed by Scientific council of Faculty Biotechnology,Meeting report __ from ______ .

Head of Scientific council of Facultyof biotechnology, PhD (in Biology) Y. V. Kolomietc

Author: V. Borodai, Associate professor of Ecobiotechnology and biodiversitydepartment

ÓБородай В.В., 2013 р.

1. Description discipline"Industrial Biotechnology"

Area of expertise, direction of training, Educational qualificationArea of expertise Biotechnologydirection of training 0514 «Biotechnology»Educational qualification «Bachelor»

Characteristics of disciplineKind of discipline regulatoryTotal hours 211The number of credits ECTS 5,8Number three content modules 3Course project (work) 63form of control Exam

Indicators discipline for full-time and distance learning

full-time learning distance learningFirst training 3 3Semester 6 6Lectures 38 4Practical, seminars - -Laboratory classes 38 6Independent work 19 131Number of weekly hoursfor full-time students:classroom:independent student work:

4 1,3

-

2. Purpose and Objectives discipline

"Industrial Biotechnology"

Industrial biotechnology formed on the base of stormy development of

molecular biology, cellular and genetic engineering, wide use of methods of

biochemistry, chemistry and other sciences. On determination of the European

biotechnological federation (EBF), a biotechnology is such integration of natural

and engineering sciences, by which the use of cages, cellular structures and

separate biomolecules enables to get more high-quality and more cheap products

of the medical and industrial setting or conduct other useful manipulations. Due to

development of medical industry there were processes of industrial receipt of

amino acid, vitamins, enzymes but other Biotechnological processes considerably

extended the sphere of production in food industry. Enzymatic catalysis replaced

by itself some reactions in the synthesis of organic compounds. Numerous

technologies of defense of environment based on the use of microorganisms and

plants, show by itself the classic examples of ecobiotechnologies.

However, without regard to the large variety of agents which carry out

biotechnological manipulations, in our time there are microorganisms a basic

object of the biotechnology. Microorganisms are used for the biosynthesis of

important metabolites (amino acid, organic acids, enzymes, exopolysaccharides,

lectins, surface active matters, vitamins, growth factors, hormones); receipt of

albuminous products, probiotics, fertilizers (preparations are on the basis of

biomass); receipt of biogas; lixiviating of metals is from ores; transformations of

matters (synthesis of steroids, ephedrine). The large enough and various group of

microorganisms is used for this purpose: micromycetes, yeasts, bacteria are

representatives of different physiology and taxonomical groups, which differ as on

the type of metabolism, and on the type of feed.

The purpose of study of discipline is a capture to knowledge and abilities of

cultivation of separate strains of industrial microorganisms students, by the

methods of selection of biological agents for the receipt of separate products,

government of cultivation of microorganisms, control of quality of the got product,

directions of application of products of biotechnology, determination of them

processes bases ecological safety, especially created on the basis of the genetically

modified microorganisms. A course foresees preparation of bachelor and leans

against knowledge of disciplines which form a specialist for different industries of

national economy, including agroindustrial production.

A task to the course is making for the students of ability to manage the

processes of cultivation of microorganism’s pilot-scale by collection, working and

analysis of information; experimental mastering of methods of work with different

industrial microorganisms in the conditions of laboratory and during educational

practices in research establishments and biochemical enterprises.

Students will get knowledge about the physiology features of industrial

strains of microorganisms, condition of receipt of products on their basis,

directions of application of such matters, and also prospects of development of

bases of industrial biotechnology in Ukraine and in the world; will lay hands on

the methods of control after the processes of biosynthesis of products on the basis

of microorganisms and estimation of degree of their risk for a human and

environment. The theoretical aspects of discipline are fastened on laboratory

employments.

Information from discipline for students is contained in textbooks, manuals,

methodical recommendations, departments developed by teachers, by scientists, in

monographs, academic and periodic editions and network of Internet.

After mastering of course a student must know ecobiotechnological bases of

plant-grower; biochemical, physiology and physical methods of analysis of

biotechnological products; devices and equipments are for biotechnological

processes; government and accident and labor protection prevention

biotechnological processes bases are ill a biotechnological laboratory, on a

production, constrained with the use of microorganisms.

A student must be able to use the obtained knowledge and skills in practice

of national economy: able to prepare nourishing environments, reagents and

tableware for sterilization, to modify them in obedience to the put tasks; to carry

out control of biotechnological operations in laboratory and production terms; to

estimate the results of biotechnological researches and processes; instrumental in

introduction of results of biotechnological researches taking into account biosafety

of environment.

Independent work is presented by the set of individual semester tasks with

the purpose of verification of mastering of educational material and methodical

pointing a student to implementation of works. Component part of control is an

estimation of knowledge on laboratory employments, during implementation of

control works.

Semester control through verification and estimation of knowledge is carried

out on the basis of module-rating systems on tests and examinations.

List of disciplines mastering of which necessity for the study of discipline

Study of discipline « Industrial biotechnology» it is based on knowledge,

what students acquire normative part of OPP at a study - cycles of preparation

humanitarian and socio-economic, naturally scientific, professional and practical

and selective part - disciplines after the choice of university and student.

A student must master thus:

1) methods of preparation of tableware and nourishing environments for

growing of living cages and culture of fabrics, biosynthesis to the albumen,

amino acid, physiological active matters, nature and many-sided nature of

biotechnological processes, prospects of development, which are studied on a

course the «General biotechnology» ;

2) structure of the immersion microscope, rule works with him,

preparation of preparations; morphology and ultra structure of cages,

physiology, genetics of microorganisms, use of their properties in a biochemical

production; diagnostics of basic microorganisms which are used in agriculture

as bacterial fertilizers and preparations for protection of plants, the methods of

their cultivation of, which are studied on a course «General microbiology».

3) diagnostics and authentication of viruses, their structure and

biochemical composition, properties and pathological influence on the plants of,

which studied on a course «Virology».

4) normative providing of biotechnological productions /STANDART /.

5) cultural plants, their feeds and development, that studied in a course

«Botanist» / procariots, eucariots-fungy, algae;

6) zoology / the simplest/

7) chemical elements and their co-operation with each other, compounds,

which are used in microbiology which is studied in a course «Chemistry»;

8) organic chemistry of /protein, carbohydrates, hydrocarbons, fats, enzymes.

3. Program of discipline "Industrial Biotechnology"

Theoretical studies

Module 1. Fermentation processes in the biotechnology industry

Topic of the lecture classes 1, 2. Using products of microorganisms in the bakery,

getting dairy products, beverages and vinegar, pickling processes and silage.

Etiological period of industrial biotechnology. Researches of L.Paster in

nature microbiological fermentation, anaerobic microorganisms living conditions,

vaccination and vaccinotherapy, methods of sterilization. Works of R. Koch,

D.Tyndalya, E. Duclos, C. Shamberlana, Mechnikov, D.Zabolotnoho and other

researchers.

Development of methods for obtaining pure cultures of microorganisms

producing food pressed yeast start commercial production of metabolic products of

microorganisms (acetone, butanol, citric and lactic acid).

Design, development and implementation of practice equipments for

industrial biotechnology, including bioreactors. Optimization of fermentation,

cultivation of microorganisms to produce practically valuable metabolites

(antibiotics, amino acids, vitamins, enzymes, etc.). The development of the food

industry. Enzymatic catalysis. Fundamental work of F. Crick and Watson (1953) to

install the structure of DNA.

Geno-technical period of industrial biotechnology. Formation of genetic

engineering, creation of the first recombinant DNA molecule. Works O.O. Baev,

A.M. Belozersky, A. Avery, G. Gamow, HG Koran, F. Jacob, J. Mono, JA

Ovchinnikov, A. Spirin, RV Petrova R.H.Butenko. Industrial production of drugs

produced by genetic engineering: insulin, interferon, human growth hormone and

others.

Topic of the lecture classes 3, 4. General characteristics of industrial strains of

microorganisms. The main ways of getting new products and strains. Screening of

new products that are synthesized industrial microorganisms. Selective allocation

of industrially-important strains. Features metabolism of microorganisms. Primary

and secondary products of anabolism and catabolism. Physiological aspects of the

regulation of metabolism strains. Population growth rate and the synthesis product.

Physic-chemical factors and synthesis of metabolites. Regulation of the synthesis

of enzymes. Requirements for industrial strains of microorganisms. Isolation

producers from natural sources. Parameters for assessing variability crops by

almost important features. Stepwise selection of industrial microorganisms.

Principles of using mutagens in the selection of microorganisms. Using mutants in

breeding resistant to analogs of metabolites, that are resistant to antibiotics, a

modified transport metabolites). Principles of mutant organisms with impaired

regulation of the synthesis of metabolites. Using genetic engineering to produce

practically valuable strains of microorganisms. Supersynthesis metabolic products

using genetic methods. Getting the proteins of humans and animals. Construction

of strains of primary and secondary metabolites. Preparations for the production

process, long-term storage and maintenance of the active state of industrial

microorganisms.

Content module II. Templates and main stage biotech industries

Theme Lectures 5,6. Classification fermentation process: breakdown by product,

main phase, with respect to oxygen and light, the degree of protection from

extraneous microflora, the number of species of microorganisms, the way the

organization. Basic parameters and phase periodic fermentation. Quantitative

characteristics of growth and productivity: growth rate, doubling time of biomass,

economic and metabolic factors influence substrate concentration on the rate of

growth, productivity and specific production rate. Kinetics of growth of

microorganisms. The curve of growth of the cell population. Quantitative

indicators of fermentation processes: formation heat of oxygen and carbon dioxide.

Types and modes fermentation: periodic and continuous processes. Problems

aeration, defaming, asepsis and sterility in various fermentation. Open and closed

fermentation system. Hemostat and turbidostat modes cultivation producers. Basic

requirements that apply to the bioreactors. Mixing systems that are used in modern

fermenters. Principles of scaling processes: laboratory, pilot and industrial

fermenters and solved using their problem. Specialized fermentation technologies:

anaerobic, solid phase and gas-phase processes.

Theme Lectures 7,8. Characteristics of the main stages of the biotech industry.

Major substrates and by-products used for microbial synthesis. Sources of carbon

and nitrogen nutrition. Natural raw materials of plant origin. Waste various

industries as raw material for biotechnological processes. Chemical and

petrochemical substrates used as raw material. Characterization of complex

medias. Requirements for growth media used in industrial biotechnology.

Preparatory stage. Preparation and sterilization of the medium, air, bioreactors,

preparation of inoculum for shallow and deep cultivation, biocatalysts, pre-

processing of raw materials. Biotech stage: fermentation, biotransformation,

biocatalysis, biookyslennya, methanogenic fermentation biokompostuvannya,

biosorbtsiya and others. Separation of liquids and biomass sedimentation,

filtration, separation, centrifugation, coagulation, flotation. Isolation internally -

and externally - cellular metabolic products: precipitation, extraction, adsorption,

electrochemical methods, ion exchange chromatography. The destruction of the

cell wall and extraction of target products. Pretreatment of culture fluid for a more

complete phase separation. Acid coagulation. Thermal coagulation. Adding

electrolytes. Purification, concentration, modification and stabilization of the target

product. Production of finished product forms. Types of products at their

destination in a typical process flow charts. Sample preparation flowchart biotech

industries

MODULE 3. Getting of biologically active substances and the individual

components of microbial synthesis

Theme Lectures 9,1011,12,13. Biotechnology products of microbial synthesis.

Classification of products of microbiological synthesis. Preparations based on

biomass of bacteria: probiotics, bacterial fertilizers, biological products for plant

protection, baking yeast protein products. Bio-based live and inactivated vaccines.

Preparations based on metabolic products of microorganisms: primary metabolites

- amino acids, organic acids, vitamines, secondary metabolites - antibiotics

exopolysaccharide, surface-active substances, etc., products of fermentation -

alcohols, organic acids, ketones; enzymes. Immobilized bacterial cells and their

applications. Methods of immobilizing cells of microorganisms. Adsorption. And

covalently cross-linking. Inclusion in the gel. Features living cells immobilized

microorganisms. Biotechnology receiving amino acids: lysine receipt, glutamic

acid, proline, leucine, tryptophan. Chemistry, Enzymology and regulation of

biosynthesis of glutamic acid, lysine, threonine, tryptophan. Methods for selection

of mutants for amino acids production. Technological schemes of amino acids by

microbiological synthesis. Getting amino acids using immobilized cells and

enzymes. Features of the enzymes of microorganisms. Enzymes microorganisms

used in production. Biotechnology enzymes: getting active producers, nutrient

media for culturing microorganisms, especially the cultivation of microorganisms

as producers of enzymes. Isolation and stabilization of enzymes. Immobilized

enzymes. The use of enzymes in the food industry, agriculture and medicine.

Nomenclature of microbial enzyme preparations. Biotechnology production of

organic acids: citric, acetic, gluconic and itakonovoyi. The use of citric acid

producers. Selection of active producers. Biosynthesis and its regulation. Terms

fermentation of Aspergillus niger. Getting citric acid industry: cultivation shallow,

deep, firmly of phase. Biosynthesis of vitamins and regulation of these processes.

Ways to get riboflavin, vitamin B12, carotenoids, ergosterol. Microbiological

synthesis of polysaccharides and lipids. Polysaccharides cytoplasm and membrane

structures. Polysaccharides of cell walls. Exopolysaccharide. The composition and

content of lipids in microorganisms. The properties of the main types of

polysaccharides and lipids of microbial origin, and their practical use. The

producers of polysaccharides and lipids. Pathways of biosynthesis. Technological

schemes of microbial polysaccharides and lipids. Industrial production of xanthan.

Microorganisms - producers of antibiotics. Pathways of biosynthesis and genetic

control of the formation of these compounds. The search for new antibiotics.

Formation of antibiotics in industrial environments. Antibiotics are formed by

bacteria, actinomycetes, filamentous fungi. Ways to improve antibiotic

biosynthesis by microorganisms. Two phase nature of antibiotic producers.

Technological foundations of antibiotics. Preparation of culture media antibiotic

producers and seed. Phase biosynthesis of antibiotics. Pretreatment culture fluid.

Isolation and purification of antibiotics. The industrial method for semi antibiotics.

Loss of the ability of microorganisms to the formation of antibiotics in industrial

environments. The using of fermentation processes and other metabolic processes

of microorganisms. Biotechnology food and fermentation products. Alcoholic

fermentation. Physiology and chemistry of yeast alcoholic fermentation.

Characterization of strains producers used in industry. Use of yeast in the industry:

Getting ethanol, production of bread. Biotechnology is a wine, beer and strong

alcoholic beverages. Lactic acid fermentation. Homo-and hetero enzymatic

fermentation. Distribution and relationship with other microorganisms. Isolation

and storage of lactic acid bacteria. Biotechnology use of lactic acid bacteria and

baker's yeast. Flow charts alcoholic and lactic fermentations. The use of lactic acid

bacteria in the process of ensiling forage, pickled vegetables, cheese, cooking

prophylactic and therapeutic drugs. Biotechnology cheese. Propionic acid

fermentation characteristics of the producers. Microorganisms used in the

fermentation industry. Acetone-butyl alcohol fermentation, especially the

cultivation of microorganisms-producers, flowsheet. Getting microbial biomass as

a source of protein. Process and principles control the cultivation of

microorganisms. The main raw materials, characteristic strains.

Theme Lectures 14,15,16. Industrial biotechnology in agriculture. Formulations

and standardization of biological products. Bacterial, viral drugs and biological

products based on fungi for plant protection. Characterization of microorganisms,

which are the basis of biological products. Entomopathogenic bacterial agents:

characterization of Bacillus thuringiensis (Bt), metabolites, the mechanism of

action of crystalline δ-endotoxin. Biotechnology receiving biologics. Bio-based

biologically active substances antagonistic microorganisms. Description of basic

biological plant protection used in Ukraine. Bacterial fertilizers and

aerbicides. Types of fertilizers and bacterial microorganisms that are used to obtain

them. Nitrogen-fixing bacterial preparation, properties rhizobia. Technological

schemes of fixing nodule bacteria, azotobakteryn, planryz, haupsyn and

tryhodermin. Inoculation and storage products. Control of production. Ways to

improve the efficiency of inoculation. Prospects of using.

4. Structure discipline "General (industrial) biotechnology"

Назви змістових модулів і тем

hoursfull-time learning distance learning

Total including Total includingLec pr lab ind ind Lec pr lab ind ind

Module 1. Enzymatic processes in the biotechnology industryTopic 1. History and achievements of industrial biotechnology. 15 6 6 3 20 2 2 16Theme 2. General characteristics of industrial strains ofmicroorganisms.

17 7 7 3 18 1 1 16

Total for module 1 32 13 13 6 38 3 3 32Content module 2. Scheme and main stage biotech industries.

Topic 1. Classification and characterization of the fermentationprocess.

15 6 6 3 20 2 2 16

Theme 2. Characteristics of the main stage biotech industries 17 7 7 3 18 1 1 16Total for module 2 32 13 13 6 38 3 3 32

Module 3. Getting biologically active substances and certain components of microbial cells.Topic 1. Biotechnology products of microbial synthesis. 16 6 6 3 17 1 1 15Theme 2. Industrial biotechnology in agriculture. 16 6 6 4 17 1 1 15

Total for module 3 31 12 12 7 34 2 2 30Total 95 38 38 19 110 8 8 94

Course project 63 45Teaching Practice (themes)

1.Types of cultivation of microorganisms-producers. 92.Enrichment cultures of microorganisms on media. 93. Cultivation of microorganisms in selective growth media. 94. Detecting microorganisms - antagonists of the biological activity

of antibiotics.9

5. Study of the effect of culture conditions on the physiologicalactivity of microorganisms-producers.

9

6. Research quality of biological preparations. 97. Introduction to the scientific advancement of microbiological

laboratory of plant protection Plant Protection Institute NAAS ofUkraine.

9

8. Introduction to the biotechnological development of the Instituteof Microbiology and Virology D.K. Zabolotny NASU.

9

Total 72General total 211

5. Topics of seminars№ Title theme Hours

1 Don’t provide curriculum

6. Topics of practical training№ Title theme Hours

1 Don’t provide curriculum

7. Title themes laboratory studies, hours

Module 1. Enzymatic processes in the biotechnology industryNumber Title theme HoursLab1,2

Principles and methods for culturing microorganisms-producers onmedia. Equipment and Materials of Laboratory of IndustrialBiotechnology. Methods of sterilization equipment and culture media

2

Lab3,4

Preparation of nutrient media for culturing bacteria and fungi in thelaboratory. Principles of nutrient media in biotech manufacturing

2

Lab5,6

Getting a batch culture Bacillus subtilis, Bacillus subtilis varmesentericus, Aspergillus niger

2

Lab7,8

Initial screening of microorganisms - antagonists in rhizosphere ofplants. Isolation of pure cultures of microorganisms - antagonists

4

Content module 2. Scheme and main stage biotech industries.Lab 11,12

Cultivation of yeast on nutrient media containing carbon substrates 5

Lab 13,14

Cultivation of microorganisms - antagonists on selective nutrientmedia

6

Lab15,16

The investigation of microbial antagonism by perpendicular strokes.Determination of the sensitivity of microorganisms to antibiotics bypaper disc

5

Module 3. Getting biologically active substances and certain components ofmicrobial cells.

Lab11,12

Determination of enzymatic activity of oxidase and catalasemicroorganisms. Cultivation of Aspergillus niger on media ofdifferent composition of macro-and micronutrients

7

Lab13,14,15

The formation of citric acid by the fungus Aspergillus niger throughcultivation on liquid medium superficial way

8

Lab 16,17,18

The efficacy of biological products against bacterial blight pathogenof plants Pectobacterium spp. Standardization and qualityassessment of biological titer determination by the drug.Standardization and quality assessment of biopharmaceuticals bydetermining the biological activity of microorganisms - producers

7

NATIONAL UNIVERSITY of LIFEand ENVIROMENTAL SCIENCE of UKRAINE

Biotechnology facultyEcobiotechnology and biodiversity department

THE CALENDAR THEMATIC PLANfrom course “"Industrial biotechnology"

For grounding of specialists in direction 6.051401- "Biotechnology"in agrarian higher educational institutions of III – IV levels of accreditation”

"APPROVED"The dean of Biotechnology faculty

_____________Y. V. Kolomietc“___” _______________ 2013

Year 3st year of Bachelor of SciencesNumber of ECTS credits – 5.8Lectures - 38 hoursLaboratory works - 38 hoursIndependent work under thesupervision of lecturer – 19 hours.Final form of control: exam

Topic of lectures Hours Topic of Laboratory works Hours. Topic of independent work under thesupervision of lecturer

Hours Literature

Module 1.Topic 1. History and achievements ofindustrial biotechnology.

2

Principles and methods for culturingmicroorganisms-producers on media.Equipment and Materials of Laboratoryof Industrial Biotechnology. Methods ofsterilization equipment and culturemedia. Preparation of nutrient media forculturing bacteria and fungi in thelaboratory. Principles of nutrient mediain biotech manufacturing

2 World famous scientists in the history ofindustrial biotechnology. Mainapplication fields of immobilized cellcultures

2

M1-4,7

Theme 2. General characteristics of industrialstrains of microorganisms.

6

Getting a batch culture Bacillus subtilis,Bacillus subtilis var mesentericus,Aspergillus niger. Initial screening ofmicroorganisms - antagonists inrhizosphere of plants. Isolation of purecultures of microorganisms - antagonists

2 Microbial kinetics Continuous stirredtank bioreactorsBubble column bioreactors Airliftbioreactors

2

A7-8

Module 2.Topic 1. Classification and characterizationof the fermentation process.

2 Cultivation of yeast on nutrient mediacontaining carbon substrates

2 Receipt of lipids, polysaccharidess, ethylspirit and organic solvents

4 M3,7.A8, 12,17.

2 Cultivation of microorganisms -antagonists on selective nutrient media

2 Synthesis of antibiotics as display ofantagonism of microorganisms

M 1,4,7.A 3,5, 6,17.

Theme 2. Characteristics of the main stagebiotech industries

2 The investigation of microbialantagonism by perpendicular strokes.

2 Bacterial and viral preparationsDetermination of bacterial contaminationof air of apartments by a sedimentationmethod with the use of universalenvironment of MPA

2

M1,5,7.A 1 1 , 12,

14

4 Determination of the sensitivity ofmicroorganisms to antibiotics by paperdisc

2 Bacteriologicexamination of milk anddairy products. Determination of quality ofmilk on content of microorganisms by themethod of dispersion on hard nutrientsmedium

M 3,7.A 8,11,14.

Module 3.

Topic 1. Biotechnology products ofmicrobial synthesis.

2 Determination of enzymatic activity ofoxidase and catalase microorganisms.Cultivation of Aspergillus niger onmedia of different composition of macro-and micronutrients.The formation ofcitric acid by the fungus Aspergillusniger through cultivation on liquidmedium superficial way

4 Determination of polysaccharidess.

M 5,7.A 8,11.

Theme 2. Industrial biotechnology inagriculture.

2 The efficacy of biological productsagainst bacterial blight pathogen ofplants Pectobacterium spp.Standardization and quality assessmentof biological titer determination by thedrug. Standardization and qualityassessment of biopharmaceuticals bydetermining the biological activity ofmicroorganisms - producers

2 Receipt of vitamins, antibiotics andenzymic preparation

4 M 6,7

LecturerResults of vertification of the calendar plan Manager of department

Cheked

NATIONAL UNIVERSITY of LIFEand ENVIROMENTAL SCIENCE of UKRAINE

Biotechnology facultyEcobiotechnology and biodiversity department

THE CALENDAR THEMATIC PLANfrom course “"Industrial biotechnology"

For grounding of specialists in direction 6.051401- "Biotechnology"in agrarian higher educational institutions of III – IV levels of accreditation”

for distance learning

"APPROVED"The dean of Biotechnology faculty

_____________Y. V. Kolomietc“___” _______________ 2013

Year 3st year of Bachelor of SciencesNumber of ECTS credits – 5.8Lectures - 6 hoursLaboratory works - 6 hoursIndependent work under thesupervision of lecturer – 94 hours.Final form of control: exam

Topic of lectures Hours Topic of Laboratory works Hours. Topic of independent work under thesupervision of lecturer

Hours Literature

Module 1.Topic 1. History and achievements ofindustrial biotechnology.

1

Principles and methods for culturingmicroorganisms-producers on media.Equipment and Materials of Laboratoryof Industrial Biotechnology. Methods ofsterilization equipment and culturemedia. Preparation of nutrient media forculturing bacteria and fungi in thelaboratory. Principles of nutrient mediain biotech manufacturing

1 World famous scientists in the history ofindustrial biotechnology. Main applicationfields of immobilized cell cultures

16

M1-4,7

Theme 2. General characteristics of industrialstrains of microorganisms.

1

Getting a batch culture Bacillus subtilis,Bacillus subtilis var mesentericus,Aspergillus niger. Initial screening ofmicroorganisms - antagonists inrhizosphere of plants. Isolation of purecultures of microorganisms - antagonists

1 Microbial kinetics Continuous stirred tankbioreactorsBubble column bioreactors Airliftbioreactors

16

A7-8

Module 2.

Topic 1. Classification and characterizationof the fermentation process.

1 Cultivation of yeast on nutrient mediacontaining carbon substrates

1 Receipt of lipids, polysaccharidess, ethylspirit and organic solvents

15 M3,7.A8, 12,17.

1 Cultivation of microorganisms -antagonists on selective nutrient media

1 Synthesis of antibiotics as display ofantagonism of microorganisms

16 M 1,4,7.A 3,5,6,17.

Theme 2. Characteristics of the main stagebiotech industries

The investigation of microbialantagonism by perpendicular strokes.

Bacterial and viral preparationsDetermination of bacterial contamination ofair of apartments by a sedimentation methodwith the use of universal environment ofMPA

8 M1,5,7.

A 1 1 ,12,14

1 Determination of the sensitivity ofmicroorganisms to antibiotics by paperdisc

1 Bacteriologicexamination of milk and dairyproducts. Determination of quality of milkon content of microorganisms by the methodof dispersion on hard nutrients medium

8 M 3,7.A 8,11,14.

Module 3.

Topic 1. Biotechnology products of microbialsynthesis.

1 Determination of enzymatic activity ofoxidase and catalase microorganisms.Cultivation of Aspergillus niger onmedia of different composition of macro-and micronutrients.The formation ofcitric acid by the fungus Aspergillusniger through cultivation on liquidmedium superficial way

1 Determination of polysaccharidess. 8

M 5,7.A 8,11.

Theme 2. Industrial biotechnology inagriculture.

1 The efficacy of biological productsagainst bacterial blight pathogen ofplants Pectobacterium spp.Standardization and quality assessmentof biological titer determination by thedrug. Standardization and qualityassessment of biopharmaceuticals bydetermining the biological activity ofmicroorganisms - producers

1 Receipt of vitamins, antibiotics andenzymic preparation

8 M 6,7

LecturerResults of vertification of the calendar plan Manager of department Cheked

8. Independent work of students

In independent work according to curriculum set aside 22 hours.Independent work of students is divided into two components - preparation forclasses and individual tasks (task description, abstract, calculation tasks, etc.).The amount of time allocated to each of these two parts are: preparation forlearning tasks - 19 hours, Individual tasks - 22 hours.

Num Title theme Hours1 Methods for cleaning up the air. Characteristics of the filter material.

Flowsheet compression and air purification. Cleaning the exhaust air.Preparation of biological agent. Keeping the original strains producers.Preparation of inoculum. Dosage and physiological condition of seed ma ¬materials. Prepare equipment and communications. Methods for detectionand quantified in the environment and man-made streams. Continuouscultivation principles of the chemostat, turbidostatu. Immobilization of cells.

4

2 Delivered by microorganisms energy. Biological oxidation. Breathing.Aerobic respiration. Krebs cycle. Respiratory electron transport chain.Incomplete oxidation of organic compounds. Anaerobic respiration. Nitraterespiration and nitrification. Sulphate and sulfur respiration. Carbonatebreathing. Fumaratne breathing. Photosynthesis. Additions of nutrients inmicrobial cell: passive transfer, active transport, phagocytosis, pinocytosis.Fermentation. The main stages of fermentation. Through the formation ofpyruvate. Types of fermentation.

4

3 The influence of environmental factors on microorganisms: humidity, theconcentration of dissolved salts, reaction (pH), temperature. Relation ofmicroorganisms to oxygen, solar radiation and anthropogenic emissions.Genotype and phenotype. Types of variability. Spontaneous mutations.Mutagenesis. Selection of microorganisms and genetic struc ¬ Neri. Transfercharacteristics and genetic recombination. Conjugation. Transduction.Transformation. Restriction and modification. Industrial strains ofmicroorganisms. Keeping microorganisms. Periodic reseeding. Storage atlow temperatures. Lyophilization. Drying. Storage in mineral oil. Effect ofstorage on microorganisms. Bacteriophages in Microbiological industry.

4

4 Regulatory and auksotrofni mutants - producers of amino acids. Vitaminsand vitamin concentrates. Vitamin B12, its production and use. Riboflavin(vitamin B2), its production and use. Obtaining and using ergosterol,carotenoids. Use of antibiotics and vitamins in medicine, agriculture andfood industry. Propionic acid fermentation. General characteristics. The useof propionic acid bacteria. Acetone-butyl alcohol fermentation. Generalcharacteristics. Bacteria that cause it. Terms cultivation. Separation offermentation products. Oxidative transformation of organic compounds:oxidation of mono, di-, tri-and polyhydric alcohols, monosaccharides.Synthesis of ascorbic acid dioksyatsetonu vinegar.

4

5 Requirements for the quality of the finished product. Food poisoning andtoxicity. General principles of microbial control of finished products in thefood industry.Microscopic control. Accounting for special environments. Sanitationcontrol equipment, materials production, hands and clothing of workers.Scheme of control, the principles of the scheme depending on the specificityof production.

6

Total 22

Self-Test Students

1. The using of microorganisms in biotechnology.2. In which areas predicted expansion of the use of biotechnology?3. Describe the growth phase of bacteria in the periodical culture.4. Parameters of the curve of growth.5. In which phase of exponential growth culture cells die faster than fall?6. What is the essence of continuous cultivation?7. What modes of continuous cultivation.8. What is bacterial mass?9. What are the fundamental differences between intermittent and

continuous cultures.10. Based on the signs of yeast fungi belong to?11. Why yeast is treated as a separate physiological groups of

microorganisms?12. What are the main stages of the study of yeast.13. What types of vegetative propagation characteristic of yeast?14. Describe the known classifications yeast.15. Describe yeast as object biotechnology.16. What are examples of yeast - Saccharomyces in the industry.17. Which substrates grow yeast Candida?18. When it was established in world production of fodder yeast?19. What is the content of lysine in proteins of different strains of yeast?20. What is fermentation?21. What are examples of using yeast-Saccharomyces in the industry.22. What microorganisms are used for commercial production of

alcoholic beverages?23. How much yeast biomass allowed to be used as food additives in

other countries?24. What are the characteristics of enzymes of microorganisms.25. What enzymes inherent microorganisms?26. What biologically active substances synthesized by fungi?27. What metabolites belonging to the primary and secondary?28. Practically valuable secondary metabolites of microorganisms.29. What is the biological importance is the ability to synthesize

antibiotics in microorganisms?30. For the biosynthesis of important metabolites practically used

microorganisms?31. What antibiotics antagonism?32. How do antibiotics on microorganisms?33. What microorganisms - producers of antibiotics.34. What methods are used to determine the antibiotics?35. What is the practical significance of microbial lectins origin?36. What do you know auksotrofni mutants?37. Which metabolic process called fermentation?38. Which organisms can carry out fermentation?

39. What fungi involved in the production of citric acid?40. What are the substrates of lactic acid bacteria produce lactic acid?41. What where the industry is widely using lactic acid.42. How to get lactic acid in industrial environments?43. In what year was opened a new era in industrial processes using

incomplete oxidation?44. What microorganisms are used for commercial production of vinegar

with natural wine?45. How much proline synthesizing bacteria genus Brevibacterium?46. What are organisms that develop in pickled vegetables.47. What is the optimum pH for the formation of citric acid?48. Describe the features of free-living nitrogen fixators, call the main

representatives of this group.49. Meaning biotechnological process of decomposition by

microorganisms phosphorus for agriculture.50. How and why organisms decompose mineral phosphates?51. Biotechnological process of extracting from the soil bacterium

fosforomobilizuyuchi?52. Describe the basis of microbiological fertilizer application.53. Why microorganisms can be used to determine the presence of soil

available to plants battery?54. What are bacterial fertilizer? Did they replace chemical fertilizers?55. What aspects and conditions of the effectiveness of bacterial

fertilizers.56. What Do You Know about bacterial fertilizer and how to use them?57. Describe the main industrial microbial polysaccharides.58. What vitamins you can get microbial synthesis?59. Describe the preparations that are based on the biomass of

microorganisms.60. What are the benefits of using biological agents to protect plants from

disease before the chemical.61. What are probiotics used for correction of the microflora of the human

body.62. What drugs to protect plants from pests63. What method is used to determine the biomass of bacteria?64. Describe water clarity with organoleptic study?65. What methods of microbial contamination of surfaces used?66. What bacteria involved in the processing of manganese ore and

aluminum?67. Due to what happens accumulation of metals?68. What fertilizer obtained by microorganisms?69. Name the bacteria that form L-glutamic acid under certain

physiological conditions?

Individual task

1. Preparing for classes

Fuller mastery of material subject "Total (industrial) biotechnology" onthe basis of assimilation analysis of literary sources in conditions of individualwork will enhance interest in specific issues of discipline. After reviewing theliterature, the student prepares a summary (descriptive task) or presentation(report) according to the indicative list of individual tasks. Summary - a reviewof recent literature (except textbooks), scientific journals, the informationoffered in the Internet. The presentation was prepared by gathering literatureand auxiliary illyustratsiynoho material. Creating a presentation in general andthe choice of forms of information are creative and individual process. Thequality of the processed sources, methodology, findings and design work intoaccount when evaluating the work.

Proposed subjects of individual tasksModule 1. Fermentation processes in the biotechnology industry.

1. Using literary sources describe the world market biotech products(descriptive work).

2. With references describe market biotech products in Ukraine(descriptive work).

3. Based on the analysis of literary sources prove prospects for use inbiotechnological processes extremal forms of microorganisms, includingthermophilic and Psychrophilic (descriptive work).

4. Based on the analysis of the literature describe methods exhaustcleaning biotech industries (descriptive work).

5. Conduct observations on the effects of cultivation conditions on thegrowth of microorganisms.

6. Using literary sources describe the most common precursors ofsecondary metabolites of biotechnological production (descriptive work).

Main literature: [ 2, с. 9-216; 3, с.3-74; 6, с.9-130; 7, с.7-136; 8, с.8-128],Additional literature: [ 10, с. 3-165, 11, с.5-232; 13, с. 1-136; 15, с.5-207;20, с. 4-213]

Modul 2. Typical schemes and main stage of biotech industries.1. Based on the analysis of the literature to describe the factors that

determine the growth and biosynthetic ability of producers (descriptive work).2. Conduct research antagonistic activity of Trichoderma viridis

shmammiv agents against major diseases of agricultural plants.3. Using literary sources describe the role of the relative concentrations of

carbon and nitrogen sources in the environment in biotechnological industries(descriptive work).

4. With references describe determinants in wastewater biotech industries(descriptive work).

5. Based on the analysis of the literature to describe the amino acids thatare formed by direct amination trans amination precursor synthesis of aminoacids (descriptive work).

6. Based on the analysis of the literature to describe the formation ofhydrocarbon precursor cell wall lipopolysaccharide in gram-negative bacteria,the synthesis of the carbohydrate component of the cell wall in eukaryotes(descriptive work).

7. With references describe acetic acid bacteria, the formation of aceticacid with ethanol to acetic acid bacteria, especially the functioning of thetricarboxylic acid cycle inherent in acetic acid bacteria (descriptive work).

Main literature: [ 1, с. 3- 256; 2, с. 236 -523; 3, с.75-198; 6, с.130-188; 7, с.136 -235; 8, с.128-278; 9, с. 322-498], Additional literature: [ 10, с. 165- 281,11, с. 232-324; 13, с. 136- 203; 16, с. 7- 120; 17, с. 40-332; 18, с. 213-502;19, с. 12-324]

Module 3. Getting biologically active substances and certaincomponents of microbial cells.

1. Using literary sources describe what features inherent synthesis ofamino acids in microorganisms (descriptive work).

2. Based on the analysis of the literature to describe ways of obtainingmutants auksotrofnyh - producers of amino acids (descriptive work).

3. Based on the analysis of the literature to describe the stages ofobtaining genetically engineered strains of E. coli - nadsyntetyka threonine(descriptive work).

4. Using literary sources describe the Krebs cycle intermediates, whichare the source for the synthesis of vitamin B12, the mechanisms regulatingbiosynthesis of vitamin B12 (descriptive work).

5. Using literary sources describe what features inherent synthesis ofdextran (descriptive work).

6. Based on the analysis of the literature to describe the reactions ofmetabolism substrate formed intermediates required for the synthesis ofpenicillins and cephalosporins (descriptive work).

7. Based on the analysis of the literature to describe the main stages of thesynthesis of macrolide antibiotics (descriptive work).

8. With references describe compounds that are the source for thesynthesis of lactone macrocycle avermektyniv, the practical importanceavermektyniv (descriptive work).

9. Using literary sources describe the features that are inherent lacticfermentation carried bifidobacteria (descriptive work).

Main literature: [ 1, с.256-698; 2, с.523-680; 3, с.198-230; 4, с. 123-312; 5, с.3-142; 6, с. 188- 290; 7, с. 235 - 397; 8, с. 278- 334, 9, с. 498-700], Additionalliterature: [11, с. 324-476; 12, с. 5-159; 13, с. 203-290; 14, с. 3-90; 17, с.332- 478; 19, с. 324 - 410].

NUBіP Ukraine Ф-7.5-2.1.8-04Structural and logіcal scheme instruction discipline

Industrial biotechnologyNumber contain

module Partdiscipline

Subject lecture Topicpractical (laboratory) studies

Shapeknowledge control

Containmodule I

History ofdevelopment ofbiotechnology

Microorganisms as objectbiotechnology.Growth of bacteria is in a periodicculture.Features of cultivation ofsaccharomycetes.Preparations onthe basis of biomass

Colorimetery method of determinationof biomass of bacterial cultureconstruction of calibration curve fordetermination of biomass.Determination of basic parameters ofgrowth of bacterial culture -2 hours.

Test

Containmodule II

Synthesis ofantibiotics as

display ofantagonism of

microorganisms

Industrial stamms of mushrooms andbacteria - producers of antibioticsBiotechnology of receipt ofpenicilirieTerms of effective application andadvantage of microbiopreparations

Microbiological control ofbakegoodss. Determination of quality of flour on thelevel of contamination microorganisms.Determination of basic indexes of thesanitary-hygenic state of surface-water.Bacterial research of tests of water oflake, river.

Test

Containmodule

III

.Generaldescription offermentation

Alcoholic fermentation.Formation of ethanol by yeasts.Formation of ethanol by bacteria.Producers, their description and use

Determination products metabolism ofbacteria.Determination of quality ofmicrobiological preparation by thelepidocid method of dispersion on hardenvironments with the next count ofcolonies.

Test

Protocolco-ordination of working educational program of discipline “Industrial biotechnology” with other disciplines for the direction of

0929 – “Biotechnology”, profession of 6.092900 – “Ecobiotechnology”, Educational – qualification level "Bachelor"

Discipline and itssections which

are beforestudying of this

discipline

Family and name,academic degree lecturer

which implementsprevious discipline

Signature

Next discipline and itsdivisions in frame of

which materials of thisdiscipline are used

Family and name,academic degree lecturerwhich implements next

discipline

Signature

Head of methodological commission of biotechnology faculty,associate Professor Y. V. Kolomietc

10. Teaching MethodsTheoretical - lectures.Practical - laboratory classes.Independent study students.

11. Forms of controlExams, Tests, final tests

12. Methods and scale assessing studentsRatings of student knowledge

Indicative distribution of points, awarded to studentstype ofcontrol

module theme Trainingsessions

(preparationand

execution)

Individual tasks modular(modulecontrol)

Total

Cur

rent

con

trol

1.1. 2 2 3 71.2. 3 3 3 9

Total 1 5 5 6 162.1. 4 4 4 122.1. 4 4 4 12

Total 2 8 8 8 243.1. 6 6 3 153.2. 6 6 3 15

Total 3 12 12 6 30Total 25 25 20 70Exam 30General total 100

Module system of teaching of discipline and a rating evaluation ofknowledge of students is after its mastering

In accordance with «Statute about credit-module system of studies in

educational material of discipline « Industrial biotechnology» it parts on 4

semantic modules, by a volume 5.8 credits of EST seach.

Rating of student from mastering of discipline is determined on a 100

ball scale. He consists of rating from educational work at the evaluation of

which 70 points target, and rating, from attestation (to examination) - 30

points. Every semantic module is also estimated on a 100 ball scale. By the

form of control of knowledges from the semantic module 1 there is

implementation of calculation work will build on transformer substation (a

task is given out to every student). Semantic module 2 estimated as a result

of defence of reports from laboratory works.

On rating from educational work in decision of department, rating can

influence from additional work - to 20 points and rating penalty (with a

negative sign) - to 5 points.

Rating of student from educational work of Rnr is determined after

a formula

Rew = 0,7(R(1)om + R(2)om)/Kdisc + Raw P - Rp, 2where R(1)OM, R(2)OM ratings estimations accordingly 1th and 2th semantic

modules after by a 100-ball by a scale;

Raw, Rp rating from additional work and rating is penal accordingly.

Students which collected from educational work 60 and more points

can not make examination, but get in examination estimation

"Automatically", in accordance with the collected amount of points,

translated in a national estimation and estimation of ESTs in obedience to a

table. 2.6. In such case rating of student from discipline R disc equals his

rating from educational work.

R disc=RewIf a student wishes to promote rating and become better estimation

from discipline, he must пройти semester attestation - to make examination.

Last students, which from educational work collected less than, than 60

points, take place in an obligatory order. For admitting to attestation a

student must collect less than 60 not points from every semaritic module,

but oh the whole not less than, than 42 points are from educational work.

Rating of student from attestation of Rat isdetermined after by a 100-

poins by a scale.

Rating of student from discipline of Rdisc is calculated on a formulaRdisc = Rew+ 0,3 RAT

Rating of student from discipline is translated in a national estimation

and estimation of ESTs

Betweens national and ESTs by estimations and rating from

discipline

Estimationnational

ОцінкаECTS

Percent ofstudents which

arrive at theproper

estimation inthe Europeanuniversities

Determination of estimationof ECTS

Rating fromdiscipline, points

Excellent A 10

Excellent - excellentimplementation is only withthe negligible quantity ofmistakes

90 -100

Good

В 25

VERY GOOD- higher middlelevel with a few errors

82-89

С 30

GOOD- in general correctwork with the certainamount of flagrant errors

74-81

Satisfactorily

D 25SATISFACTORILY - notbad, but with the far offailings

64-73

E 10

SUFFICIENTLY -implementation satisfiesminimum criteria 60-63

Unsatisfactorily

FX

UNSATISFACTORILY - it isneeded to work before to get atest (positive estimation) 35-59

F ■

UNSATISFACTORILY -thesubsequent thoughtful piece ofwork is needed

01-34

A national estimation is written down in the test book of student, and an

estimation of ESTs a book of rating estimation of knowledges of student.

Intermediate control of knowledge of students is carried out regularly onlecture and practical employments by their questioning from passed material.Form of control of knowledges from the semantic module 1 is implementationof calculation work from the calculation of ground will build on transformersubstation. Semantic module 2 estimated as a result of defence of reports fromlaboratory works.

Final control of knowledges is carried out on examination.An estimation is "excellent proposed" to the student which during a

semester worked systematic, on examination rotined scalene and deepknowledges of programmatic material, able freely to execute tasks which areforeseen by the program, mastered basic arid acquainted with additionalliterature, intercommunication of separate sections of discipline, their value feelsfor a future profession, found out creative capabilities in understanding and useeducational-programmatic to material, showed a capacity for an independentupdate and addition to knowledges.

An estimation is "Good proposed" to the student which found outcomplete knowledge educational-programmatic to material, successfullyexecutes the foreseen program tasks, mastered basic literature which isrecommended by the program, proof character of knowledges rotined fromdiscipline and apt at their independent addition and renewal during subsequentstudies and professional activity.

An estimation is "Satisfactorily proposed" to the student whichdiscovered knowledge basic educational-programmatic to material in a volume,to the necessity for subsequent studies and next work after a profession,produced with implementation of tasks, foreseen by the program, assumedseparate errors in an answer on examination and at implementation ofexamination tasks, but owns necessary knowledges for their overcoming underthe direction of scientifically pedagogical worker.

An estimation is "Unsatisfactorily proposed" to the student which did notfind out sufficient knowledges basic educational-programmatic to material,assumed of principle errors in execution the tasks foreseen by the program, itdoes not can unassisted scientifically pedagogical to use a worker knowledge atsubsequent studies, not able to lay hands on skills of independent work.

13. Methodology Support1. Guidelines for the study of the subject "Industrial Biotechnology" forbachelors direction 6.051401 "Biotechnology" distance learning.2. Guidelines for the study of the subject "Industrial Biotechnology" forbachelors direction 6.051401 "Biotechnology" for independent studycourses.3. Guidelines for the study of the subject "Industrial Biotechnology" forbachelors direction 6.051401 "Biotechnology" - a course of lectures.4. Guidelines for writing a term paper on the subject "IndustrialBiotechnology" for bachelors direction 6.051401 "Biotechnology".

14. Course project

Course project summarizes the development of the discipline "Total(industrial) biotechnology."

The main objectives of the course are:

- develop skills of independent work, deepening the theoretical andpractical knowledge;

- organization and analysis of the literature of the studied issues;

- formulation of scientific and practical tasks, the choice of methodsfor its implementation and development of the project design;

- scientific analysis of the problems raised;

- choice of ways to implement design solutions for specificproduction conditions.

The general requirements for course work are:

- The conditions of production;

- logic structure and presentation of the material;

- concise language;

- convincing arguments and evidentiary findings.

The structure consists of course work;

1. Introduction.

2. Presentation of the material.

3. Conclusions.

The introduction should briefly assess the current state of the question,novelty and relevance of the work.

When providing the basic material required to analyze the generalcharacteristics of industrial strains of microorganisms, principles and methods ofcultivation of microorganisms-producers on artificial nutrient media inlaboratory and industrial conditions, a template and fundamental stages biotechindustries. Based on this analysis to make recommendations regarding theoptimal conditions for conducting basic biotechnological processes to producebiologically active compounds, principles and methods of constructing objectsbiotechnology cultivation of certain strains of industrial microorganisms,methods of selection of biological agents in order to obtain certain products,management of cultivation bases microorganisms and control the quality of theproduct. This is reflected in the findings.

Protecting the course work is conducted in the form of a short report andanswers to questions that arise as a result of studying the work.

Indicative list of topics course projects

1. Modern industrial biotechnology in Ukraine and in the world (2000-2013).

2. Microbial strain selection (induced mutagenesis) in the industrialbiotechnology

3. Recombinant DNA Technology, protoplasts fusion and Improvement ofIndustrial Microorganisms

4. Principles, general design and types of bioreactors and fermenters.5. Types and stages of fermentation process6. Different types of purification methods in the industrial biotechnology7. Biotechnology of Yeast Biomass Production8. Production of antibiotics by Actinomycetes (Streptomyces spp.)9. Production of antibiotics by Molds (Penicillium spp.)10.Microbial production of Acetic Acid11.Lactic acid fermentation. Microbial production of Lactic acid.12.Microbial production of primary and secondary metabolites13.Microbial production of Citric Acid14.Alcoholic beverage biotechnology industry. Biotechnology of brewing

and winemaking15.Variety and potential applications microbial exopolysaccharides in the

industrial biotechnology16.Biotechnological production of bio-fertilizers on the basis of

phosphate-mobilizing bacteria17.Biotechnological production of bio-fertilizers on the basis of nitrogen-

fixing bacteria.

18.Biotechnological production of plant protection biopreparations (PGPR –bacteria)

19.Biotechnological production of plant protection entomopathogenicbiopreparations (Bacillus thuringiensis)

20.Exploring the biotechnological applications in the Archaeal Domain

15.Teaching practice studentsTeaching practice students is a continuation of the training process in

industrial biotechnology laboratories. During practice student broadens and

deepens the theoretical knowledge in industrial biotechnology laboratories. The

main purpose of the practice is deepening and consolidation of acquired

knowledge and skills gained during the study of theoretical course discipline and

a relevant practical skills in the study of cultivation conditions of industrial

strains and stages biotech industries including classical and modern scientific

approaches. Target practice is to develop students' skills of general principles

and methods of cultivation of microorganisms, a batch culture of bacteria and

fungi, which are used in biotechnological production, assembly typical patterns

and fundamental stages biotech industries, methods of obtaining biologically

active substances and certain components of microbial cells, experimental

development of methods to work with different objects in a biotech lab and

research farms.

As a result of the practice of student fully masters the knowledge and

skills of Industrial Biotechnology:

1. Technique cultivation of microorganisms-producers on artificial

media in the laboratory. Students reinforce knowledge learned in the study

subjects, sterilized equipment, tools, and culture media. Preparation of nutrient

media for culturing bacteria and fungi in the laboratory. Principles of Mediums

in biotech manufacturing.

2. Getting a batch culture of microorganisms on the media. Conduct

initial screening of microorganisms - antagonists in the rhizosphere of plants.

Isolation of microorganisms methods flushing and dilution and seeding method

on selective nutrient media. Obtaining pure cultures of microorganisms. Getting

a batch culture Bacillus subtilis, Bacillus subtilis var mesentericus, Aspergillus

niger.

3. Cultivation of microorganisms on selective media. Cultivation of

yeast on nutrient media containing carbon substrates. Cultivation of

microorganisms - antagonists on selective media. Obtaining seed (inokulyuma)

for metabolic products.

4. Identification of microorganisms - antagonists biological activity of

antibiotics. The study obtained strains on antagonistic activity by perpendicular

strokes. Determination of the sensitivity of microorganisms to antibiotics using

paper discs. Determination of the sensitivity of microorganisms to phytoncids.

5. Influence of cultivation conditions on the physiological activity of

microorganisms-producers. Determination of enzymatic activity of oxidase

and catalase microorganisms. Investigation of changes in pH, mass Aspergillus

niger mycelium when cultured on media of different composition of macro-and

micronutrients. Getting citric acid by the fungus Aspergillus niger through

cultivation on liquid medium superficial way.

6. Research of biopreparations. Determination of the effectiveness

against biological pathogens of crops. Assessment of the quality of biological

titer determination by the drug and biological activity of microorganisms -

producers.

7. Introduction to microbiological laboratory method of plant

protection Institute of Plant Protection NAAS of Ukraine. Introduction to

laboratory equipment and techniques used to produce biopharmaceuticals.

Biotechnology is a microbiological preparations for plant protection.

8. Introducing the biotech developments Institute of Microbiology

and Virology of D.K. Zabolotnyi. Familiarization with the methods and

principles to develop new probiotics and biologics to protect plants from

bacteria of the genus Bacillus and Pseudomonas in the department of antibiotics,

advanced techniques reidentyfikatsiyi nokardio and korynepodibnyh bacteria,

lactic acid bacteria and yeast in the department of physiology of

industrial microorganisms.

Students are working practice diaries, which regularly record information

about their work: the knowledge, to participate in seminars, tours, workshops,

and do not understand the question, which clarifies the head of practice. The

results of students write in worksheets. Report practice is to measure the

accumulation of materials.

As a result of the practical training trainee receives credit. The basis for

classification is the presentation to the department the following documents:

- Blog educational practice;

-Report traineeship, signed by the supervisor of practice.

Evaluation of credit entered into the record information and academic

records of the student.

16. Educational - methodical materials from discipline

Main:1. Машины и аппараты пищевых производств : учебник для

вузов/ [Антипов С.Т., Кретов И.Т., Остриков А.Н. и др. ]; подред. В.А. Панфилова. - М.: Высшая школа, 2001. - 704 с. – (в 2 кн.,кн.1).

2. Промышленная микробиология : учеб. пособие для вузов /[Аркадьева З.А., Безбородов А.М., Блохина И.Н. и др. ]; под ред.Н.С. Егорова. - М.: Высшая школа, 1989. - 688 с.

3. Безбородов А.М. Биотехнология продуктов микробного синтеза:Ферментативный катализ, какальтернатива органического синтеза /А.М. Безбородов - М.: Агропромиздат, 1991. – 238 с.

4. Бекер М.Е. Биотехнология / Бекер М.Е., Лиепиньш Г.К., РайпулисЕ.П.– М.: Агропромиздат, 1990. – 334 с.

5. Егоров Н.С. Биотехнология : микробиологическое производствобиологически активных веществ и препаратов: учеб. пособиедля вузов. / Н.С. Егоров, В.Д. Самуилов. - М.: Высшая школа,1997. – 143 с. – ( в 8 кн., кн.6).

6. Воробъёва Л.И. Промышленная микробиология: учеб.пособие /Воробъёва Л.И. – М.: Изд-во МГУ, 1989. – 294 с.

7. Пирог Т.П. Загальна мікробіологія: підручник / Пирог Т.П. — К.:НУХТ, 2004. — 471 с.

8. Пирог Т.П. Загальна біотехнологія: підручник / Т.П. Пирог, О.А.Ігнатова. – К.: НУХТ, 2009. – 336 с.

9. Сельскохозяйственная биотехнология / [Шевелуха В.С.,

Калашникова Е.А., Кочиева Е.З. и др. ]; под ред. В.С.Шевелухи. – [3-е изд., перераб. и доп.] – М.: Высшая школа, 2008. – 710 с.

Additional:10.Антипова Л.В. Прикладная биотехнология: Учеб. пособие для вузов /

Антипова Л.В., Глотова И.А., Жаринов А.И. - Воронеж: Воронеж.гос. технол. акад., 2000.- 283 с.

11.Биотехнология. Принципы и применение / [Бич Г., Бест Д., БрайерлиК. и др. ] Пер. с англ. ; под ред. И. Хиггинса, Д. Беста, Дж. Джонса. –М.: Мир, 1988. – 480 с.

12.Биотехнология микробных ферментов / [Лобанок А.Г., АстаповичН.И., Михайлова Р.В., Безбородов А.М.] – Минск: Наука и техника,1989. – 204 с.

13.Бирюков В. В. Основы промышленной биотехнологии / Бирюков В.В. — М.: КолосС, 2004. — 296 с.

14.Витол И.С. Экологические проблемы производства и потребленияпищевых продуктов: учеб.пособие. / Витол И.С. - М.: МГУПП, 2003.– 93 с.

15.Микробиология / [Воробьев АА., Быков A.C., Пашков Е.П.,Рыбакова A.M. ] — М.: Медицина. — 1998. — 336 с.

16.Голубев В.Н. Пищевая биотехнология / В.Н.Голубев, И.Н. Жиганов.– М.: Делипринт, 2001.– 123 с.

17.Елинов Н.П. Основы биотехнологии: Для студентов институтов /Елинов Н.П. – СПб: Наука, 1995. – 600 с.

18.Кавецкий Г.Д., Васильев Б.В. Процессы и аппараты пищевойтехнологи / Г.Д. Кавецкий, Б.В. Васильев. - М.: Колос, 2000.- 551 с.

19.Неверова О.А. Пищевая биотехнология продуктов из сырьярастительного происхождения/ О.А. Неверова, Г.А.Гореликова,В.М.Позняковский. – Новосибирск: Сиб.унив.изд-во, 2007. – 415 с.

20.Современная микробиология. Прокариоты / Под ред. И. Ленгелера,Г. Древса, Г. Шлегеля. — М.: Мир, 2005. — Т. 1. — 654 с.

17. Information Resources

1. http://www.biotechnolog.ru2. http://fp7-bio.ru/biotech/nkt-in-russia3. http://www.processindustries.mwgroup.net/ru/glavnaja

Summary of lectures (short version) by discipline "Industrial Biotechnology"

Module 1. Fermentation processes in the biotechnology industry

Topic of the lecture classes 1, 2. Using products of microorganisms in the

bakery, getting dairy products, beverages and vinegar, pickling processes and

silage. Etiological period of industrial biotechnology. Researches of L.Paster in

nature microbiological fermentation, anaerobic microorganisms living

conditions, vaccination and vaccinotherapy, methods of sterilization. Works of

R. Koch, D.Tyndalya, E. Duclos, C. Shamberlana, Mechnikov, D.Zabolotnoho

and other researchers. Development of methods for obtaining pure cultures of

microorganisms producing food pressed yeast start commercial production of

metabolic products of microorganisms (acetone, butanol, citric and lactic acid).

Design, development and implementation of practice equipments for industrial

biotechnology, including bioreactors. Optimization of fermentation, cultivation

of microorganisms to produce practically valuable metabolites (antibiotics,

amino acids, vitamins, enzymes, etc.). The development of the food industry.

Enzymatic catalysis. Fundamental work of F. Crick and Watson (1953) to install

the structure of DNA. Henotehnichnyy period of industrial biotechnology.

Formation of genetic engineering, creation of the first recombinant DNA

molecule. Works O.O. Baev, A.M. Belozersky, A. Avery, G. Gamow, HG

Koran, F. Jacob, J. Mono, JA Ovchinnikov, A. Spirin, RV Petrova

R.H.Butenko. Industrial production of drugs produced by genetic engineering:

insulin, interferon, human growth hormone and others.

Topic of the lecture classes 3, 4. General characteristics of industrial strains of

microorganisms. The main ways of getting new products and strains. Screening

of new products that are synthesized industrial microorganisms. Selective

allocation of industrially-important strains. Features metabolism of

microorganisms. Primary and secondary products of anabolism and catabolism.

Physiological aspects of the regulation of metabolism strains. Population growth

rate and the synthesis product. Physic-chemical factors and synthesis of

metabolites. Regulation of the synthesis of enzymes. Requirements for industrial

strains of microorganisms. Isolation producers from natural sources. Parameters

for assessing variability crops by almost important features. Stepwise selection

of industrial microorganisms. Principles of using mutagens in the selection of

microorganisms. Using mutants in breeding resistant to analogs of metabolites,

that are resistant to antibiotics, a modified transport metabolites). Principles of

mutant organisms with impaired regulation of the synthesis of metabolites.

Using genetic engineering to produce practically valuable strains of

microorganisms. Supersynthesis metabolic products using genetic methods.

Getting the proteins of humans and animals. Construction of strains of primary

and secondary metabolites. Preparations for the production process, long-term

storage and maintenance of the active state of industrial microorganisms.

Content module II. Templates and main stage biotech industries

Theme Lectures 5,6. Classification fermentation process: breakdown byproduct, main phase, with respect to oxygen and light, the degree of protectionfrom extraneous microflora, the number of species of microorganisms, the waythe organization. Basic parameters and phase periodic fermentation.Quantitative characteristics of growth and productivity: growth rate, doublingtime of biomass, economic and metabolic factors influence substrateconcentration on the rate of growth, productivity and specific production rate.Kinetics of growth of microorganisms. The curve of growth of the cellpopulation. Quantitative indicators of fermentation processes: formation heat ofoxygen and carbon dioxide. Types and modes fermentation: periodic andcontinuous processes. Problems aeration, defaming, asepsis and sterility invarious fermentation. Open and closed fermentation system. Hemostat andturbidostat modes cultivation producers. Basic requirements that apply to thebioreactors. Mixing systems that are used in modern fermenters. Principles ofscaling processes: laboratory, pilot and industrial fermenters and solved usingtheir problem. Specialized fermentation technologies: anaerobic, solid phase andgas-phase processes.

Theme Lectures 7,8. Characteristics of the main stages of the biotech industry.Major substrates and by-products used for microbial synthesis. Sources ofcarbon and nitrogen nutrition. Natural raw materials of plant origin. Wastevarious industries as raw material for biotechnological processes. Chemical and

petrochemical substrates used as raw material. Characterization of complexmedias. Requirements for growth media used in industrial biotechnology.Preparatory stage. Preparation and sterilization of the medium, air, bioreactors,preparation of inoculum for shallow and deep cultivation, biocatalysts, pre-processing of raw materials. Biotech stage: fermentation, biotransformation,biocatalysis, biookyslennya, methanogenic fermentation biokompostuvannya,biosorbtsiya and others. Separation of liquids and biomass sedimentation,filtration, separation, centrifugation, coagulation, flotation. Isolation internally -and externally - cellular metabolic products: precipitation, extraction,adsorption, electrochemical methods, ion exchange chromatography. Thedestruction of the cell wall and extraction of target products. Pretreatment ofculture fluid for a more complete phase separation. Acid coagulation. Thermalcoagulation. Adding electrolytes. Purification, concentration, modification andstabilization of the target product. Production of finished product forms. Typesof products at their destination in a typical process flow charts. Samplepreparation flowchart biotech industries

MODULE 3. Getting of biologically active substances and the individualcomponents of microbial synthesis

Theme Lectures 9,1011,12,13. Biotechnology products of microbial synthesis.Classification of products of microbiological synthesis. Preparations based onbiomass of bacteria: probiotics, bacterial fertilizers, biological products for plantprotection, baking yeast protein products. Bio-based live and inactivatedvaccines. Preparations based on metabolic products of microorganisms: primarymetabolites - amino acids, organic acids, vitamines, secondary metabolites -antibiotics ekzopolisaharydy, surface-active substances, etc., products offermentation - alcohols, organic acids, ketones; enzymes. Immobilized bacterialcells and their applications. Methods of immobilizing cells of microorganisms.Adsorption. And covalently cross-linking. Inclusion in the gel. Features livingcells immobilized microorganisms. Biotechnology receiving amino acids: lysinereceipt, glutamic acid, proline, leucine, tryptophan. Chemistry, Enzymology andregulation of biosynthesis of glutamic acid, lysine, threonine, tryptophan.Methods for selection of mutants for amino acids production. Technologicalschemes of amino acids by microbiological synthesis. Getting amino acids usingimmobilized cells and enzymes. Features of the enzymes of microorganisms.Enzymes microorganisms used in production. Biotechnology enzymes: gettingactive producers, nutrient media for culturing microorganisms, especially thecultivation of microorganisms as producers of enzymes. Isolation andstabilization of enzymes. Immobilized enzymes. The use of enzymes in the foodindustry, agriculture and medicine. Nomenclature of microbial enzyme

preparations. Biotechnology production of organic acids: citric, acetic, gluconicand itakonovoyi. The use of citric acid producers. Selection of active producers.Biosynthesis and its regulation. Terms fermentation Aspergillus niger. Gettingcitric acid industry: cultivation shallow, deep, firmly of phase. Biosynthesis ofvitamins and regulation of these processes. Ways to get riboflavin, vitamin B12,carotenoids, ergosterol. Microbiological synthesis of polysaccharides and lipids.Polysaccharides cytoplasm and membrane structures. Polysaccharides of cellwalls. Exopolysaccharides. The composition and content of lipids inmicroorganisms. The properties of the main types of polysaccharides and lipidsof microbial origin, and their practical use. The producers of polysaccharidesand lipids. Pathways of biosynthesis. Technological schemes of microbialpolysaccharides and lipids. Industrial production of xanthan. Microorganisms -producers of antibiotics. Pathways of biosynthesis and genetic control of theformation of these compounds. The search for new antibiotics. Formation ofantibiotics in industrial environments. Antibiotics are formed by bacteria,actinomycetes, filamentous fungi. Ways to improve antibiotic biosynthesis bymicroorganisms. Two phase nature of antibiotic producers. Technologicalfoundations of antibiotics. Preparation of culture media antibiotic producers andseed. Phase biosynthesis of antibiotics. Pretreatment culture fluid. Isolation andpurification of antibiotics. The industrial method for semi antibiotics. Loss ofthe ability of microorganisms to the formation of antibiotics in industrialenvironments. The using of fermentation processes and other metabolicprocesses of microorganisms. Biotechnology food and fermentation products.Alcoholic fermentation. Physiology and chemistry of yeast alcoholicfermentation. Characterization of strains producers used in industry. Use ofyeast in the industry: Getting ethanol, production of bread. Biotechnology is awine, beer and strong alcoholic beverages. Lactic acid fermentation. Homo-andhetero enzymatic fermentation. Distribution and relationship with othermicroorganisms. Isolation and storage of lactic acid bacteria. Biotechnology useof lactic acid bacteria and baker's yeast. Flow charts alcoholic and lacticfermentations. The use of lactic acid bacteria in the process of ensiling forage,pickled vegetables, cheese, cooking prophylactic and therapeutic drugs.Biotechnology cheese. Propionic acid fermentation characteristics of theproducers. Microorganisms used in the fermentation industry. Acetone-butylalcohol fermentation, especially the cultivation of microorganisms-producers,flowsheet. Getting microbial biomass as a source of protein. Process andprinciples control the cultivation of microorganisms. The main raw materials,characteristic strains.

Theme Lectures 14,15,16. Industrial biotechnology in agriculture.Formulations and standardization of biological products. Bacterial, viral drugs

and biological products based on fungi for plant protection. Characterization ofmicroorganisms, which are the basis of biological products. Entomopathogenicbacterial agents: characterization of Bacillus thuringiensis (Bt), metabolites, themechanism of action of crystalline δ-endotoxin. Biotechnology receivingbiologics. Bio-based biologically active substances antagonisticmicroorganisms. Description of basic biological plant protection used inUkraine. Bacterial fertilizers and herbicides. Types of fertilizers andbacterial microorganisms that are used to obtain them. Nitrogen-fixing bacterialpreparation, properties rhizobia. Technological schemes of fixing nodulebacteria, azotobakterynu, planryzu, haupsynu and tryhoderminu. Inoculation andstorage products. Control of production. Ways to improve the efficiency ofinoculation. Prospects of using.

Form № Н-5.05NATIONAL UNIVERSITY of LIFE and ENVIROMENTAL SCIENCES of UKRAINE

Faculty BiotechnologyEducational qualification «Bachelor»Direction of training 0514 «Biotechnology»Form of education Full-time, distance learningSemester, course 6 semester, course 3Discipline Industrial biotechnology

«Approved»Ecobiotechnology and biodiversity department

«___» ____________ 2013Head of department____________O. L. Klyachenko

Lecturer – Borodai V.V.

Examination card №Question 1. Xanthomonas campestrisbacteria used in industrial biotechnologyfor the production…………..

(Fill in the form correct answer)Question 2. Move the target productfrom the aqueous phase in a nonmixable with water organic matter:1. flotation2. coagulation3. extraction4. dialysis

Question 3. Find the accordancebetween two columns:

Question 4. What methods are used todetermine the activity of antibiotics inthe lab?1 The method of paper discs2 method of Vinogradsky3 The method of agar blocks4 The method of calculation in the

chamber Horyayeva5 Method of perpendicular strokes

Question 5. Sources of carbon nutritionare:

1 molasses, hydrological, starch,ammonium sulfate

2 starch, hydrological, lactose, corn flour,methyl alcohol

3 ammonium sulfate, ammonium nitrate,urea, hydrol

4 ammonium nitrate, urea, corn extractQuestion 6. Preparation of catalyticagent refers to the stage:

1 Concentration of the product2 Separation of liquid and biomass3 Purification4 Biotech stage5 Preparation stage

Question7. Mechanism of action of β-lactam antibiotics

1 mechanism of action is different2 disturbance of synthesis of cell wall3 irreversible inhibition of protein

synthesis4 reversible inhibition of protein synthesis5 violation of plasma membrane

Question8. Over- synthesis of citric acid ispossible at ...

1 limitation the producer of K and Mg,excess sucrose, low pH

2 limitation the producer of P and Fe,excess sucrose, low pH

3 limitation the producer of N and C, anexcess of ammonium sulfate, pH neutral

4 limitation the producer of Zn and S, anexcess of ammonium sulfate, high pH

Question9. Autotrophs used as sourcesof carbon:А lipidsБ carbohydratesВ organic matterГ amino acids and proteinsД carbon dioxide

A. manycyclical process

1. loading raw and inoculumoccurs simultaneously, afterfermentative liquid unloaded

В. continuousprocess

2. A part of fermentativeliquid material in theapparatus serves as theinoculum for furtherfermentation at discharge

С. periodicprocess

3. loading and unloading ofmedia takes place at thesame time for a long time

Question10. By the universal physicalmutagens include:fill in the form of answers correct answerQuestion11. Which the metabolicproducts related to primary andsecondary products of catabolism andanabolism?

A. Primaryproducts ofanabolic

1. Lipids, vitamins,biomass,polysaccharides andcellular biosynthesisintermediates:nucleotides and aminoacids

B. Secondaryproducts anabolic

4. Acetone, butanol.

V. PrimaryProductscatabolism

3. Ethanol, acetic acid,CO2, ATP.

G. Secondaryproductscatabolism

4. Antibiotics,alkaloids, toxins.

Question12. What is the correctsequence of phases of cell populationgrowth of bacteria?

1. The lag phase2. Acceleratedgrowth phase3. The logarithmicphase4. Phase of slowgrowth5. The stationaryphase6. The deathphase

Question13. Rapid freezing attemperatures from -35 to -78 ° C, transfervials into a vacuum-drying apparatus anddrying under a pressure 13,33-1,33 Pawithin 25-30 h, following sealing vials ofdried cultures in an atmosphere of inert gasis called ... ...Fill in the form of answers correct answerQuestion14. The main advantage ofmicrobial synthesis of amino acids is toget them ... ... ..Fill in the form of answers correct answer

Question15. The name of process ofprotein biosynthesis is………

1 transcription2 replication3 translation4 transformation5 restriction

Question 16. For adsorption ... ... containsprotein molecules on the surface of thecarrier by electrostatic, hydrophobic,dispersion interactions and hydrogen bondsFill in the form of answers correct answerQuestion17. Chemical factors mutagenicaction:Fill in the form of answers correct answerQuestion 18. The trophophase includes:

1 Accelerated growth phase2 The stationary phase3 The death phase4 The logarithmic phase5 Phase of slow growth6 The lag phase

Question19. High-purity enzymesare:….

1 10х

2 20х

3 3хQuestion 20. Cell membranes ofbacteria built with:1) protein and carbohydrates;2) lipids and proteins;3) cellulose;4) nucleic acids and lipids.

Question21. The most common aminoacids, obtained by microbiologicalsynthesis, are:

1 tryptophan, dextran, proline, lysine,surfaktyn

2 proline, tsiankobalamid, lysine, glutamicacid, leucine, tryptophan

3 glutamic acid, leucine, tryptophan,dextran, proline, lysine

4 proline, lysine, glutamic acid, leucine,tryptophan