12 Industrial Fermentation 2008 Ho

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Industrial fermentation Production with microbes Industrial microbiology

The use of microbes in the production

Start from an ancient time

Large scale production

Early 20th century beverage industry, vinegar,

baking yeast, citric and lactic acid

Late 20th century development of

biotechnology from the antibiotic industrial

Production with microbes

Fermentation industry

Microbes to produce useful product

Usually a naturally produce substances for

growth, and maintenance.

Metabolites

Primary metabolite produce during growth

Secondary metabolite produce after growth

stage

Substrate

Product

Cell

Time

Substrate

Product

Cell

Time

Primary metabolite

Secondary metabolite

Trophophase Idiophase

ProductsI. Foods and foods related- Fermented meat, Cheeses and milk products,mushrooms, bakers yeast, coffee, pickles, singlecell-proteins, vinegar, amino acids, vitamins,alcoholic beverages

ProductsII. Organic acids- Citric acids, Itaconic acid

III. Enzymes and microbial transformation

- Commercial enzyme, sterol conversion

IV. Engineered microbes

- Insulin, human growth factor


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Production with microbes

Steps in Industrial fermentation

Isolation of microbes that produce your

interested product

Screen for the best producing strain: naturally or

mutation.

Optimize production condition (growth

basically) in lab

Scale up from lab scale (up to 10 L) to industrail

scale (>10,000 L) (raw material?)

Food and food related products

Whole cells products

Including mushrooms and Single-cell protein

(SCP) Mushrooms fruiting body of mold!

Highly consumed every year high nutrition.

Grow on decaying organic matter in soil or wood.

Stalk and cap = compact mycelium

Gills = site of reproductive spores

E.g. Agaricus bisporus(botton mushroom),

Lentinus edulus(Shitake mushroom)

Agaricus bisporusFood and food related products

Single-cell protein yeast, and cyanobacteria

Alternative food sources mostly used now as

supplementary diet e.g. yeast is a source for B

Vitamins.

Organisms: yeast, Spirulina

Growing biomass and freeze dry Raw material from other industrial, farming


Bakers yeast Same process as in single cell protein

Food and food related products Cheese production

Can produce from any milk.

4 phases: coagulation, separation, shaping

and ripening

Coagulation: rennin enzyme or acids precipitatecasein

From calvess stomachs, engineered bacteria

Acid production by lactic acid bacteria

Curd entrap bacteria, fat globules and othermaterial.


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Cheese production

Separation & Shaping

Additives (salt and herbs) may be added after seperation

or shaping: effect property of cheese product

Ripening add flavor and look

Natural or inoculation of microbes e.g. Penicillium

roquefortii (spores are usually added into blue

cheese), and Penicillium camembertiis added to

the surface of a cheese known as Camenbert

Blue cheese

Brie

Gorgonzola

Swiss


Yogurt and other fermented milk products

Yogurt, kefir, buttermilk, sourcream

Yogurt usually use mixture of 2 cultures:

Streptococcus thermophilus: ini. acid production

Lactobacillus bulgaricus

0.5 - 1.0% lactic acid

Fermented at 28-32 C

Taste due to acetaldehyde, diacetyl, and acetoin

Product need to contains live cultures

A mixed lactic acid and

alcoholic fermentation.

Streptococcus lactis,

Lactobacillus

bulgaricus, yeasts

Kefir

This milk product is used for its

medicinal therapeutic value.

Lactobacillus

acidophilusAcidophilu

s milk

Made with skimmed or partly

skimmed pasteurized milk.

Streptococcus sp.

Leuconostoc sp.Cultured

buttermilk

Cream is inoculated and leave

to develop acidity

Streptococcus sp.

Leuconostoc sp.Sour cream

DescriptionMicroorganismsFermented

Product

Some Foods Produced from Fermented Milk.

Industrial production

of yogurtKefir Buttermilk


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Alcoholic beverages

Accidental discovered from ancient human

Saccharomyces cerevisiae Raw materials are regional.variety of products

Wine grapes, fruits

Beer cereal grain barley, sorgum, wheat

Other rice wine (rice), teguila (agave cactus),

vodka (corn), mead (honey).


Fermented meat

Use to preserve food E.g. salami, summer

sausage, cured ham

Thailand naam (fermented pork), fermented

sausage, fermented fish, fish sauceetc

Main organism Pediococcus serevisiae,

Lactobacillus plantarum, other lactic acid bacteria

Production of lactic acid to change pH of food

Production of probiotics in some strains


Fermented fruits and vegetables

Found around the world

With bacteria, yeast and mold

Depends mainly on native microbes

associate with the plants.

Most included mixed culture

fermentation

Conditioning by human to favor growth

of beneficial microbes - salting

http://www.coffeeresearch.org/Pictures/fermentationtanks.jpg

https://reader009.{domain}/reader009/html5/0525/5b07df5ad6cbb/5b07df5ee7adb.jpg

Rhizopus oligosporus, Rhizopus oryzae,

Lactobacillus delbrueckii, Saccharomyces

rouxii

Soy sauce

Leuconostoc mesenteroides,

Saccharomyces marscianus,

Flavobacterium spp., Fusarium spp.

Coffee

Lactobacillus mesenteroids, Lactobacillus

plantarum

Sauerkraut

Pediococcus cerevisiae, Lactobacillus

plantarum

Pickles

Leuconostoc mesenteroides, Candida sp.Cocoa products

OrganismsProducts Food and food related products Flavoring agents: amino acids and nucleotides

Monosodium glutamate, inosinic acid flavor

enhancers

Originally use seaweed, fish

Produce by enzymatic hydrolysis of yeast RNA

(Candida utilis)

Now direct fermentation = monosodium glutamate

and 5-IMP (inosine monophosphate)

Brevibacterium ammoniagenesproduce inosine and

then chemically phosphorylated


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Non-food products

Enzyme production

Produce in large scale around the world

Major producer Novozyme (44%), GenencorIntl, USA (18%)

Cheese productionMicrococcusLipases

Fruit juice clarificationAspergillusPectinase

Cheese manufactureAlcaligenes,

Aspergillus,Candida

Rennin

Starch liquefaction, brewing,

baking, feed, detergents

BacillusBacterial amylases

Fructose syrup productionAspergillusGlucoamylase

Antitumor agentEscherichia,

Serratia

Asparaginase

SweetenersBacillus, StreptomycesGlucose isomerase

Semisynthetic penicillinsEscherichiaPenicillin acylase

DetergentsBacillus, StreptomycesBacterial proteases

UseOrganismsEnzyme Non-food product

1. Vitamins 2. Antibiotics

3. Alkaloids 4. Steroids

5. Non-Steroid Hormones/cell regulators (cytokines):

1. Epidermal growth factor 2. Proinsulin

3. Insulin 4. Human growth hormone

5. Somatostatin 6. Interferons

7. Platelet-derived growth factor

8. Fibroblast growth factor 9. Tumor Necrosis Factor

10. Other cytokines are coming on line all the time

Non-food product6. Blood coagulating factor XIII

7. The restriction enzymes

8. Other enzymes (e.g. ligase, DNA polymerases etc.)

9. Solvents

How do the microbes produce these products? Genetically modified microorganisms

Clone genes that encode protein of interest into

vectors and express this genes in microbes used

in production.

Able to produce high amount of compounds than

in original organisms with ease of manipulation.

Microbes able to utilize waste from other process

as a raw material for fermentation.


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Antibiotics

Penicillin by Penicillium chrysogenum

Now most of the antibiotics were discover from soil

microorganisms especially in Streptomyces spp. Engineered genes into production strains.

Modification of existing antibiotics.

http://www.sanger.ac.uk

http://scienceblogs.com

Streptomycin from Streptomyces griseus

http://www.agen.ufl.edu/~chyn/age4660/lect/lect_21/f15_11.JPG

Alkaloids

Nitrogen containing organic compounds that

derived from plants

Have medicinal purposes:

Atropine dilates pupils of the eyes (some muscle

disease)

Morphine and codeine relief of pain

Cocaine local anesthetic

Quinine, caffeine, nicotine, strychnine, serotonin , and

LSD

Atropineserotonin

Steroids Types of products

include cortisone

and reproductive

hormones

Human proteins


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Biotech Drugs

Mainly recombinants proteins and monoclonalantibodies.

Highly active fields of biotech

Lots of new products each year

For list of product approve in US www.bio.org

Timeline in drug development

Documents

12 Industrial Fermentation 2008 Ho