BS2011 2/2008

Production with microbes

Industrial fermentation „ 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 Cell

Product
„ Fermentation industry
‰ Microbes to produce ‘useful’ product Secondary metabolite
‰ Usually a naturally produce substances for Cell
growth, and maintenance. Substrate Product

‰ Metabolites
Time
„ Primary metabolite – produce during growth
„ Secondary metabolite – produce after growth Primary metabolite
Substrate
stage
Trophophase Idiophase

Time

Products
I. Foods and foods related
- Fermented meat, Cheeses and milk products,
mushrooms, baker’s yeast, coffee, pickles, single
cell-proteins, vinegar, amino acids, vitamins,
alcoholic beverages
Products
II. 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 Food and food related products

„ 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) Æ in lab
„ 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

‰ Scale up from lab scale (up to 10 L) to industrail ‰ E.g. Agaricus bisporus (botton mushroom),
scale (>10,000 L) (raw material?) Lentinus edulus (Shitake mushroom)

Food and food related products

Agaricus bisporus
„ 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

Food and food related products Food and food related products
„ Baker’s yeast
‰ Same process as in single cell protein „ Cheese production
‰ Can produce from any milk.
‰ 4 phases: coagulation, separation, shaping
and ripening
„ Coagulation: rennin enzyme or acids precipitate
casein
‰ From calves’s stomachs, engineered bacteria
‰ Acid production by lactic acid bacteria
‰ Curd entrap bacteria, fat globules and other
material.

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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 camemberti is added to
the surface of a cheese known as Camenbert

Blue cheese Food and food related products

„ Yogurt and other fermented milk products
‰ Yogurt, kefir, buttermilk, sourcream

Brie
‰ Yogurt usually use mixture of 2 cultures:
„ Streptococcus thermophilus: ini. acid production
„ Lactobacillus bulgaricus
‰ 0.5 - 1.0% lactic acid
Gorgonzola
‰ Fermented at 28-32 °C
‰ Taste due to acetaldehyde, diacetyl, and acetoin
‰ Product need to contains live cultures
Swiss

Some Foods Produced from Fermented Milk.

Fermented Microorganisms Description

Product
Sour cream Streptococcus sp. Cream is inoculated and leave
Leuconostoc sp. to develop acidity
Cultured Streptococcus sp. Made with skimmed or partly
Leuconostoc sp.
buttermilk skimmed pasteurized milk.
Acidophilu Lactobacillus This milk product is used for its
s milk acidophilus medicinal therapeutic value.
Kefir Streptococcus lactis, A mixed lactic acid and
Lactobacillus alcoholic fermentation.
bulgaricus, yeasts
Industrial production Kefir Buttermilk
of yogurt

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Food and food related products

„ 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).

Food and food related products Food and food related products
„ Fermented meat „ Fermented fruits and vegetables
‰ Use to preserve food E.g. salami, summer ‰ Found around the world
sausage, cured ham ‰ With bacteria, yeast and mold
‰ Thailand – naam (fermented pork), fermented ‰ Depends mainly on native microbes
sausage, fermented fish, fish sauce…etc associate with the plants.
‰ Main organism – Pediococcus serevisiae, ‰ Most included mixed culture
Lactobacillus plantarum, other lactic acid bacteria fermentation
‰ Production of lactic acid to change pH of food ‰ Conditioning by human to favor growth
‰ Production of probiotics in some strains of beneficial microbes - salting

http://www.coffeeresearch.org/Pictures/fermentationtanks.jpg
http://www.sfu.ca/geog351fall03/groups-webpages/gp8/prod/fregment.jpg

Food and food related products

Products Organisms
„ Flavoring agents: amino acids and nucleotides
Sauerkraut Lactobacillus mesenteroids, Lactobacillus
plantarum ‰ Monosodium glutamate, inosinic acid – flavor
enhancers
Pickles Pediococcus cerevisiae, Lactobacillus
plantarum ‰ Originally use seaweed, fish
Soy sauce Rhizopus oligosporus, Rhizopus oryzae, ‰ Produce by enzymatic hydrolysis of yeast RNA
Lactobacillus delbrueckii, Saccharomyces (Candida utilis)
rouxii
‰ Now direct fermentation = monosodium glutamate
Coffee Leuconostoc mesenteroides,
and 5’-IMP (inosine monophosphate)
Saccharomyces marscianus,
Flavobacterium spp., Fusarium spp. ‰ Brevibacterium ammoniagenes produce inosine and
Cocoa products Leuconostoc mesenteroides, Candida sp. then chemically phosphorylated

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Non-food products
„ Enzyme production
‰ Produce in large scale around the world
‰ Major producer – Novozyme (44%), Genencor
Intl, USA (18%)

Enzyme Organisms Use Non-food product

Bacterial proteases Bacillus, Streptomyces Detergents
1. Vitamins 2. Antibiotics
Asparaginase Escherichia, Antitumor agent
Serratia 3. Alkaloids 4. Steroids
Glucoamylase Aspergillus Fructose syrup production 5. Non-Steroid Hormones/cell regulators (cytokines):
Bacterial amylases Bacillus Starch liquefaction, brewing,
1. Epidermal growth factor 2. Proinsulin
baking, feed, detergents
Glucose isomerase Bacillus, Streptomyces Sweeteners 3. Insulin 4. Human growth hormone
Rennin Alcaligenes, Cheese manufacture 5. Somatostatin 6. Interferons
Aspergillus,
7. Platelet-derived growth factor
Candida
Pectinase Aspergillus Fruit juice clarification 8. Fibroblast growth factor 9. Tumor Necrosis Factor
Lipases Micrococcus Cheese production 10. Other cytokines are coming on line all the time
Penicillin acylase Escherichia Semisynthetic penicillins

Non-food product How do the microbes produce these products?

6. Blood coagulating factor XIII „ Genetically modified microorganisms
7. The restriction enzymes „ Clone genes that encode protein of interest into
8. Other enzymes (e.g. ligase, DNA polymerases etc.) vectors and express this genes in microbes used
9. Solvents 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

„ 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

Streptomycin from Streptomyces griseus

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

„ Steroids „ Human proteins

‰ Types of products
include cortisone
and reproductive
hormones

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„ Biotech Drugs
‰ Mainly recombinants proteins and monoclonal
antibodies.
‰ 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

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