CHEM-C2310 Bioprocess Technology Spring 2021

CHEM-C2310 Bioprocess Technology Spring 2021

Bioprocess technology - Introduction


Tero Eerikäinen• 1979 Helsinki University of Technology (TKK)• 1980-1981 Military service in navy• 1986 Master of Science • 1989 Licentiate in Technology • 1993 Doctor of Technology • 1991-1992 Detmold Germany; research scienti• 1989-1998 TKK, laboratory manager• 1998-2003 TKK, Senior assistant• 2003-2015 TKK, Senior lecturer• 2015-2016 Aalto, Research fellow• 2016-2019 Aalto, University teacher• 2020… Aalto, University lecturer

Bioprocess Technology CHEM-C2310

• After completing the course the student will be able to:

• Describe biotechnology as an industrial sector and products and methods thereof, including applications in different fields and challenges and opportunities of industrial biotechnology

• Present transition phenomena in bioprocesses (material and energy) qualitatively and quantitatively and estimations of the behavior of biological components (cells, proteins, substrates, products, enzymes) under process conditions

• Identity the aseptic requirements in equipment and process design and applications thereof

• Present the concept of joining unit operations into bioprocesses and identify phenomena thereof and factors influencing such phenomena

• Formulate basic process models for cell and enzymatic processes based on material and energy balance and differential equations

• Name key variables of bioprocesses and describe the measuring technologies thereof as well as mathematic solutions and basics of process controls


Bioprocess technology news

Content covered in the course• Cells: operation in reactors, kinetics and quantification of growth and yield, dynamic

phenomena, process implementation methods and related simple mathematical solutions, basics of metabolism.

• Bioreactors: typical solutions, transfer phenomena, scale-up / scale-down design and research methods, asepsis in reactor design, measurement and control engineering solutions.

• Enzymes: enzymes as industrial products, equilibrium reactions, practical kinetics, thermodynamics of reactions, operation in reactors, immobilized enzymes and effects of mass transfer at the principle level.

• Processes: basics of typical unit operations, utilization of material and energy balances, consideration of biological factors in process design.

• Fundamentals of mathematical modeling of phenomena, operations and processes, with special regard to biocomponents.



Study materials & course completion

• E-book: Pauline M. Doran: Bioprocess engineering principles, 2nd edition, 2012, where applicable; readable from the link: http://libproxy.aalto.fi/login?url=http://www.sciencedirect.com/science/book/9780122208515

• Lecture and Exercise materials• MyCourses assignments • Grade formation:

– exam: 75 % – assignments: 25%

http://libproxy.aalto.fi/login?url=http://www.sciencedirect.com/science/book/9780122208515

Bioprocess technology in Finnish (help / supplementary material)


Bioprocess technology Spring 2021 program

Week Day Date Time Lecture(green)/Exercise (white) Doran's chapter9 Tuesday 02/03/2021 10:15-11:45 1. Introduction 1;3

Thrursday 4.3. 12:30-14:00 Exercise 1 (growth kinetics) 3;1210 Tuesday 9.3. 10:15-11:45 2. Cell as production unit 3;4;5

Thrursday 11.3. 12:30-14:00 Exercise 2 (stoichiometry bioprocesses) 411 Tuesday 16.3. 10:15-11:45 3. Cultivation methods 14

Thrursday 18.3. 12:30-14:00 Exercise 3 (mass balances) 412 Tuesday 23.3. 10:15-11:45 4. Bioreactors 14

Thrursday 25.3. 12:30-14:00 Exercise 4 (chemostat) 1413 Tuesday 30.3. 10:15-11:45 5. Aseptic working 9;14

Thrursday 1.4. 12:30-14:00 Exercise 5 (sterilization) 12;1414 Tuesday 6.4. 10:15-11:45 6. Mass transfer 7;8;10

Thrursday 8.4. 12:30-14:00 Exercise 6 (kLa calculations) 1015 Tuesday 13.4. Exam week

Thrursday 15.4.16 Tuesday 20.4. 10:15-11:45 7. Unit operations in bioprocesses 11

Thrursday 22.4. 12:30-14:00 Exercise 7 (filtration) 11Tuesday 11.5. 13:00-16:00 Course exam



Biotechnology combines science and engineering so that organisms, cells, cell parts or molecular analogues can be applied to the production of products and services.

Biotechnology

Bioprocess technologyThe bioprocess is based on the utilization of biocatalysts.Bioprocess technology researches, develops and applies bioprocesses.

The colors of biotechnology

Red biotechnology vs. white biotechnology

Plant investment and operating costs!


How does biotechnology affect us?

• The benefits of biotechnology include, for example:

• healing of infectious diseases, advanced diagnostics

• creating more environmentally efficient chemicals and fuels

• increasing crop yields • Disadvantages of biotechnology

include, for example• antibiotic resistant

bacteria• new allergic reactions• seed costs for farmers



Protein from carbon dioxide in the air and sunlight:https://edition.cnn.com/2020/01/20/europe/solar-foods-solein-scn-intl-c2e/index.html?fbclid=IwAR1KmgqSyFo_9C_tkYdns8JPPJhcDyyxupPruoJobjd3DN7VKdgHgo2HqaQ

History, penicillin production:https://www.youtube.com/watch?v=UTfLs8EJe20


https://edition.cnn.com/2020/01/20/europe/solar-foods-solein-scn-intl-c2e/index.html?fbclid=IwAR1KmgqSyFo_9C_tkYdns8JPPJhcDyyxupPruoJobjd3DN7VKdgHgo2HqaQ

https://www.youtube.com/watch?v=UTfLs8EJe20

Bioreaction Engineering Principles

Cell/enzyme kinetics

Unstructured Structured

Steady-state balances

Stoichiometry

Scale-up

Energetics

MixingIdeal reactor

Reaction rates

Mass transfer

CATALYST PERFORMANCE

BIOREACTOR PERFORMANCE

CALCULATION TOOLS

Modified from: Villadsen, Nielsen, Liden:Bioreaction Engineering Principles, 3rd Ed. 2011

• Combining engineering computing and life sciences

• Quantitative analysis

Chemist engineer vs. bioengineer?


Biotechnology as a production technology• Bulk products vs. fine chemicals• Host organisms

• Microbes• Plant cells• Mammalian cells• Insect cells

• Finland is also known for its enzyme industry


When the biotechnological process?

• The biotechnological process often has tocompete with other methods

• The choice is decided, for example, by – price– product complexity– environmental friendliness

Production process

What has enabled the development of biotechnological processes?

Nutrient mixing, sterilization and

pre-cultures

Recovery and

purification

Down stream processBioreactionPretreatment

Bioprocess development work

Translated from http://www.finbio.net/download/biotech-week_2013/jari-vehmaanpera.pdf


Microbiology Gene cloning Microbial technology Enzyme technology

Protein chemistryProtein chemistry Enzyme

technology

Analytics

AnalyticsBioinformatics

Application research

Application research

BiodiversityProtein

engineering Strain development Process optimization

Enzyme products

Formulation

Environmental samplesStock collectionsGenomics mining

VectorsPlasmids

PromotersAntibodies

Cell factories-Trichoderma-Bacillus-Aspergillus

Bioreactors-laboratory reactors-pilot scale reactors

Intensive screeningAutomation

RoboticsColony picker

FACS

Developed mutant linesIndustrial property rightsGenetic engineering tools

Fluid handling robots

DNA chipsProteomics

Fermented products Yeasts, lactic acid bacteria 1 000 Mt/a beer, wine, dairy products600 000 M€/a

Ethanol S. cerevisiae 90 Mt/a chemical, beverage, fuel60 000 M€/a

Antibiotics fungi, bacteria 20 000 t/a antimicrobial drugs(Streptomyces spp.) 25 000 M€/a

Vitamines bacteria, fungi 150 M€/a drugs, feed, food(B12,B2,C)

Amino acids bacteria 2 Mt/a food, feed(MSG,Lys,Thr) (Corynebacterium spp.) 2500 M€/a

Organic acids bacteria, fungi 3 Mt/a food, chemicals, polymers2000 M€/a

Thickeners bacteria 0,1 Mt/a food, cosmetics, oil drilling

Enzymes fungi, bacteria 3300 M€/a detergents, feed, (yeasts) food, pharmaceuticals,

Therapeutic yeasts, bacteria, > 120 000 M€/a pharmaceuticalsproteins mammalian cells

Cell mass S. cerevisiae,starters 0,5 Mt/a foods

Some products of cellular processesProduct Producer Production Application

L-lysine biosynthesis


Would you make L-lysine by fermentation or synthetically?

Corybacterium glutamicum L-lysine

Lactic acid bacteriaNisin

Propionibacterium sp. B12-vitamin

Aspergillus nigerfungus α-amylase

Some fermentation products

• Simple, non-chiral compounds (history, security, economics)• “Small” chiral molecules• Complex organic compounds• Polymers with a specific structure and exact composition

(enzymes, peptides, other proteins, polysaccharides)

Acetobacter sp. Acetic acid CH3COOH


http://www.ph.ed.ac.uk/%7Edra/images/acetic_l.gif

http://ligand.genome.ad.jp:8080/compound/query.html?dbkey=compound&keyword=C02823

Products of enzymatic processes

Glucose syrup α-amylase, gluco amylase, 12 Mt/a (dw) foodspullulanase 4000 M€/a

Maltose syrup α-amylase, β-amylase foods

HFCS e.g. glucose syrup + 8 Mt/a (dw in USA) foods glucose isomerase 3000 M€/a (USA)

Aspartame Thermolysin (protease) 20 kt/a foods 500 M€/a (intensive sweetener)

Acrylamide Nitrile hydratase 150 000 t/a* chemicals (polyacrylamide)500 M€/a

Psicose D-tagatose-3-epimerase foods (low-calorie sweetener

Product Enzyme Production Application

* biotechnical prod.dw = dry weightBioprocess technology - Introduction


ASPECTS RELATED TO THE CALCULATION AND PRESENTATION OF THE DATA



Figure 3.6

Growth kinetics of unicellular organisms

)(12

)(12

12

12

)()()()(tai

))0()(ln( )0()(

ttµ

ttµ

tµ

etNtNetXtX

tµX

tXeXtX

−

−

×

⋅=

⋅=

⋅=⋅=µ = specific growth rateWhen the cells grow exponentially, µ = constanti.e. then µ ≠ f(t)

X(t): cell concentration at time tN(t): number of cells at time t



Figure 3.8FIGURE 3.8 Transformation of constant errors in y after (a) taking logarithms or (b) inverting the data. Errors in ln y and 1/y vary in magnitude as the value of y changes even though the error in y is constant.

Calculate the error range for ln(y), when error for y is ± 0,5 and the range for y is 5…15:

Error bars represent a constant error in y, in this case equal to B/2


Figure 3.9

Two frequently occurring nonlinear functions are the power law and the exponential function. These relationships are often presented using graph paper with logarithmic coordinates.

Logarithmic plot (log-log-plot)


Figure 3.11

Semi-log-plot

Figure 3.10

Logarithmic scale and graph

Semi-logarithmic scale and graph


Process based on fermentation

Upstream Production Downstream

In the production of therapeutic proteinsup to 80-90% of production costs

-Block diagram of the production process-Design of pilot plant equipment-Complement the process with material balances-Examine the effect of process conditions-Calculate material and energy balances-Calculate the quantities of commodities Bioprocess technology - Introduction

Raw-materials Pre-cultivation

Inoculumn

water

Cell separation Purification

Plantcultivation

Sterilization

Operating commodities

AirCooling water

Waste or side product

product

An example of a fermentation-based process

Vacuum drum filterUltrafiltration unit

Growing in a petri dish Inoculations (here are laboratory fermentors) Production fermentors

(about 1…500 m3)

Packaged product


Examples• BioPharmax implementation of an insulin factory:• https://www.youtube.com/watch?annotation_id=annotation_3392797453&fe

ature=iv&src_vid=-_LCGuK3Q1g&v=JCA_jr19FrM

• GE: KUBio modular biopharma facility built in Germany, assembled in China

• https://www.youtube.com/watch?v=JEDhsPpD4Xo

• GE: FlexFactory™ Single-use Bioprocess Platform• https://www.youtube.com/watch?v=k39LYWnofk8


https://www.youtube.com/watch?annotation_id=annotation_3392797453&feature=iv&src_vid=-_LCGuK3Q1g&v=JCA_jr19FrM

https://www.youtube.com/watch?v=JEDhsPpD4Xo

https://www.youtube.com/watch?v=k39LYWnofk8

34Bioprocess technology - Introduction

Figure 3.14

FIGURE 3.14 Process flow sheet showing the major operations for production of bacitracin.

35Bioprocess technology - Introduction

Figure 3.15FIGURE 3.15 Quantitative flow sheet for the downstream processing of 2,3-butanediol based on fermentation of 1000 bushels of wheat per dayby Aerobacillus polymyxa.

• Corn-derived starch is converted into D-glucose

• The glucose liquor feed is fed in a down-flow manner into a series of fixed bed IGI reactors

• D-glucose syrup is converted into a HFCS-42 mixture containing

• Chromatographic enrichment to HFCS-90

• HFCS-90 blended with HFCS-42 to produce HFCS-55

Enzymatic process for high fructose corn syrup (HFCS)


Enzymatic processes

• Amylolytic enzymes are cheap => used as single batches without recycling and recovery; some are also destroyed (inactivated) in the process

• Glucose isomerase is slightly more expensive => immobilized on a solid support and can be used in columns (pictured) for several months



Bioprocesses from the perspective of a production engineer

• minimum/ complex• pH, T, aeration• batch,.. continuous• rates• measure and calculate• mixing, mass transfer • dimensions, scale-up• on-, offline, physical, chemical…• black-box, …• recovery, purification• separation, mixing…

• development of culture medium• cultivation conditions• cultivation technology• growth kinetics• substance balances• flow dynamics• reactor characteristics• measurement and control• modeling• dowstream processing• unit operations

Documents

CHEM-C2310 Bioprocess Technology Spring 2021