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Fermentative Production of Vitamins:
    Indian and Global Scenario

                          Irfan Ahmad
                 M. Tech. Pharm. Tech. (Biotech.)
National Institute of Pharmaceutical Education and Research, S.A.S. Nagar
CONTENTS
 1.   Introduction to Vitamins.
 2.   Industrial Production of Vitamins.
 3.   Vitamins Market Scenario.
 4.   Fermentative Production of Water Soluble Vitamins.
         i.   Vitamin B₁₂
         ii. Vitamin B₂
         iii. Vitamin C
         iv. Vitamin H
 5.   Fermentative Production of Fat Soluble Vitamins.
         i.   Vitamin E
         ii. Vitamin K
 6.   Summary.
Vitamins
Vitamins are defined as essential micronutrients that are
required in trace quantity and cannot be synthesized by
mammals but are essential for metabolism of all living
organisms
                        VITAMINS



Fat Soluble Vitamins                 Water Soluble Vitamins
Vitamin A                           Vitamins B
Vitamin D                           Vitamin C
Vitamin E                           Vitamin H
Vitamin K                           Vitamin P, etc.
Role of Vitamins

1   Nutritional and Physiologic Role
                   .
2        Food/Feed Additives


3         Therapeutic Agents


4            Health Aids


5           Technical Aids
Industrial Production Of Vitamins




      Extraction   Chemical   Fermentation
Industrial Production of Vitamins
                    Production Method                       World Production (tonne)
                    Biotech.       Chem.       Extr.         1980      1990      2000      2010
Thiamine (B1)                         +                      1700 4200 3700
Riboflavin (B2)         +                                    2000 2400        4400
Niacin                                +          +           8500 22000 2800
Pantothenic acid                      +          +           5000 7000
Pyridoxine (B6)                       +                      1600 2550 3800
Biotin                  +             +                      2.7   2.5    88  112
Folic acid                            +                      100   400  534
Vitamin B12             +                                    2      10    25  175
Vitamin C               +             +                     40000 60000 10700
Vitamin A                             +          +          16800 2500 2700
Vitamin D                             +          +                 5000
Tocopherol (E)          +             +          +          6800 22000 30000
Vitamin K                             +                             500




               Survase SA, Bajaj IB, Singhal RS: Biotechnological production of vitamins
Microbial Production of Vitamins
Vitamin                                 Enzyme (Microorganism)

Vitamin C     2,5-diketo-D-gluconic acid reductase (Corynebacterium sp.).
              ucu
Biotin        Fermentation (Serratia marcescens); Multiple enzyme system
              (Bacillus sphaericus).

Riboflavin    Fermentation (Eremothecium ashbyii, Ashbya gossypii,
              Bacillus sp.).
              Fermentation (Propionibacterium shermanii, Pseudomonas
Vitamin B12
              denitrificans)

Vitamin E     Freshwater microalgae, Euglena gracilis.


              Shimizu S: Vitamins and related compounds: microbial production
Fermentative Production of Vitamins

Advantages                              Disadvantages
  i.      Efficient,
                                        i.   Complex,
  ii.     Less Energy Intensive,
  iii.    Carried out at ambient        ii.  Require Careful Monitoring,
          temperature and pressure,     iii. Limited Operations Region,
  iv.     Produce Renewable             iv.  Susceptible to Substrate and
          byproducts (e.g., Biomass),        Product Inhibition,
  v.      Low Cost of Waste Disposal,
                                        v. High Cost of Downstream
  vi.     Highly enantioselective and        Processing,
          regioselective,
  vii.   Don’t suffer from consumer     vi. High Risk of Contamination,
         consciousness with regard to   vii. Tendency to Elicit Allergic
         safety.                             Manifestations.
Vitamin Market Scenario
• Europe represents the largest regional market; the US
  constitutes the single largest market globally.
• Asia-Pacific is likely to emerge as the fastest growing market,
  with a CAGR of about 4.0% over the analysis period.
• Vitamin E represents the largest segment, owing to the
  extensive use of these vitamins in cosmetics, pharmaceuticals,
  and food end-use applications
• The cosmetics industry, though relatively small in terms of
  the percentage share, is emerging as a key end-user with a
  CAGR of 4.6% over the analysis period.
• The global vitamin market was $2.3 billion in 2000.
  However, the vitamin market is expected to reach US$3.2
  Billion by 2017.
Top Five Sources of US Vitamin Imports
                                          China   Belgium   Other   Germany   India   Netherland



The vitamin industry was once
dominated by a core group of
                                                            3% 3%
producers in the developed economies                 7%

but is now far more open and             13%
competitive as an increasing number
of manufacturers in emerging
                                                                                              57%
economies such as China and India
                                         17%
weigh in.
Hoffmann La Roche
(Switzerland),     BASF
(Germany),          ADM
(USA), Hubei Guangji
(China),           Merck
(Germany) and Takeda
(Japan) are some of the
global leaders in vitamin
production.
Sales
                                                     Single vitamin
In 2011, the Indian vitamin segment                      0.3%
grew 7.5 per cent in volumes (against
                                                   Single
0.1 decline in the year 2010) and 10.3             Mineral
per cent in value (against 5.4 per cent             9%
                                          Tonics
growth in 2010).By the end of              12%
2014, the vitamins and minerals
category will be worth INR
21,174.6m ($486.7m), with an
expected CAGR of 3% between 2009
and 2014.
                                                                      Multivitamins
                                                                          79%
Five of the top 10 players are foreign
multinational firms. Among Indian
companies, the largest        vitamin
producers are Wockhardt, Raptakos
Brett, Piramal Healthcare and Alkem.
WATER SOLUBLE VITAMINS

        Vitamin B₁₂
        Vitamin B₂
        Vitamin C
        Vitamin H
Vitamin B₁₂( Cyanocobalamin)

 Cyanocobalamin is the industrially produced
  stable cobalamin form which is not found in
  nature.
 It is obtained exclusively by fermentation
  process.
 Merck began production of vitamin B₁₂ by
  Pseudomonas denitrificans in 1952 and have
  improved the efficiency of culture more than 30-
  fold relative to the performance of the original
  soil isolates by genetic manipulations and
  microbial screening.
 Mutagenic treatments have resulted in improved
  activity.




                             Sumi: Microbial Production of Vitamin B12
Vitamin B₁₂
Biosynthesis of Vitamin B₁₂
Downstream Process of Vitamin B₁₂
Vitamin B2 (Riboflavin)
• Riboflavin has been produced commercially by chemical
  synthesis, by fermentation and by a combination of
  fermentation and chemical synthesis.
• Recently, fermentation route has been widely used as it
  produces the vitamin in a single step, resulting in substantial
  cost    savings.     In contrast, chemical processes are
  multistage, and incur a lot of cost.
• Most           of         the          producers           like
  BASF, Roche, ADM/Aventis, Hubei Guangji prefer
  fermentative production of riboflavin over chemical process.
Vitamin B₂
• Although bacteria (Clostridium sp.) and yeasts (Candida sp.) are good
  producers, two closely related ascomycete fungi, Eremothecium ashbyii
  and Ashbya gossypii, are considered the best riboflavin producers.
  Ashbya gossypii produces 40000 times more vitamin than it needs for
  its own growth.
• Recently, Nippon Roche, Japan, has developed and commercialized a
  single step fermentative riboflavin production using a recombinant
  Bacillus subtilis strain, which effectively produces riboflavin directly
  from glucose in fed-batch operation.
• Improved strains for the production of riboflavin were constructed
  through metabolic engineering using recombinant DNA techniques in
  Corynebacterium ammoniagenes.
• Sybesma et al. developed Lactococcus lactis strain using both direct
  mutagenesis and metabolic engineering for simultaneous overproduction
  of both folate and riboflavin.
Vitamin C (Ascorbic Acid)
• L-ascorbic acid finds its use mainly in food industry, being a
  vitamin as well as an antioxidant.
• Majority of commercially manufactured L-ascorbic acid is
  synthesized via Reichstein process using D-glucose as a
  starting material
• Approximately 50 % of synthetic ascorbic acid is used in
  vitamins supplements and pharmaceutical preparations.
• Because of its antioxidant properties and its potential to
  stimulate collagen production, it is also widely used as an
  additive to cosmetics.
• The current global market of L-ascorbic acid is in excess of
  US$ 585 million with an annual growth rate of 3 %.
Vitamin C (Ascorbic Acid)
.
                         D-Glu

            D-Sorbitol



                          Reichestein Process
    L-Sorbose
                                                   L-
                                                Ascorbic
                Diacetone-L-      2-Keto-L-       acid
                  Sorbose         Gluconic
                                 acid methyl
                                    ester
Bacterial Fermentation of Vitamin C
                              2-Keto-D-gluconic acid
Sorbitol Pathway
                                     Pathway
Yeast Based Fermentation Process


• Saccharomyces        cerevisiae      and Zygosaccharomyces
  bailiiaccumulate produce L-ascorbic acid intracellularly
  when incubated with L-galactose.
• Over-expression of the D-arabinose dehydrogenase and D-
  arabinono-1,4-lactone oxidase in Saccharomyces cerevisiae
  enhances this ability significantly.
Algae Based Fermentation Process
• Skatrud and Huss described a method that involved initial
  growth of Chlorella pyrenoidosa ATCC53170 in a fermentor
  with a carbon source that is sufficient for the cells to grow
  to an intermediate density. At the depleted stage, additional
  carbon source was added sequentially or continuously to
  maintain      the   carbon source concentration below a
  predetermined level until the addition is terminated. This
  resulted in the production of 1.45 g/L of L-ascorbic acid.
• Euglena gracilis Z. is one of the few microorganisms
  which simultaneously produce antioxidant vitamins such
  as carotene (71 mg/L), vitamin C (86.5 mg/L) and vitamin E
  (30.1 mg/L).
Vitamin H (Biotin)
 Biotin (vitamin H) is one of the most fascinating cofactors
  involved in central pathways in pro- and eukaryotic cell
  metabolism.
 While humans and animals require several hundred micrograms
  of biotin per day, most microbes, plants and fungi appear to be
  able to synthesize the cofactor themselves.
 Biotin is added to many food, feed and cosmetic
  products, creating a world market of 10–30 t/year.
 Majority of the biotin sold is synthesized chemically via
  Goldberg and Sternbach syntheses.
 The chemical synthesis is linked with a high environmental
  burden, much effort has been put into the development of biotin-
  overproducing microbes
Biosynthesis of Biotin



                                     bioF gene



                                     bioA gene


                                     bioD gene

The conversion of
                                     bioB gene
dethiobiotin to biotin
has not been resolved.
Fermentative Production of Biotin
• Ogata et al. screened microorganisms and demonstrated that
  the bacterium B. sphaericus can excrete significant quantities
  of    biotin   synthetic     pathway    intermediates   from
  precursor, Pimelic acid.
• Brown et al. studied the production of biotin by recombinant
  strains of E. coli. The strain used was E. coli C268 having a
  bio A genotype and plasmid pTG3410 with two expression
  cassettes .
• Biotin production under limiting growth conditions by
  Agrobacterium/Rhizobium HK4 transformed with a modified
  Escherichia coli bio operon.
FAT SOLUBLE VITAMINS


       Vitamin E
       Vitamin K
Vitamin E
 Most abundant among fat soluble vitamins and has the
  highest antioxidant activity in vivo.
 In nature, only photosynthetic organisms are capable of
  producing α-tocopherol.
 In humans, ∞-tocopherol is believed to play a major role in
  prevention of light induced pathologies of the skin, eyes and
  degenerative disorders such as atherosclerosis, cardiovascular
  diseases and cancer.
 Industrial application of ∞-tocopherol includes its use in
  preservation of food, in cosmetics and sunscreens.
Vitamin E

                                   Extraction


                                                     The synthetic form is a
                                                     mixture of eight stereoisomers
Extraction from oils is not
                                                     collectively known as all-rac-
efficient, as these typically
                                                     alpha-tocopherol.
contain low levels of               Vitamin E
∞-tocopherol                                         when administered in equal
                                                     amounts, the bioavailability of
                                                     natural with respect to
                                                     synthetic ∞-tocopherol is 2:1.


                                Chemical Synthesis

                                Chemical Synthesis
Vitamin E
Fermentative Production of Vitamin K₂
   Microalgae Euglena gracilis Z and marine microalgae
    Dunaliella        tertiolecta produce ∞-tocopherol in
    concentrations higher than conventional foods.
   Carballo-Cardenas et al. studied ∞-tocopherol production
    with       Dunaliella      tertiolecta   and     Tetraselmis
    suecica, demonstrating that nutrient composition can be used
    as a tool to improve α-tocopherol productivity.
Vitamin K₂
 Vitamin K₁
                   .
              Vitamin K   Vitamin K₂
Vitamin K

Fermentative Production of Vitamin K₂
 Tani and Taguchi have reported that as much as 182 mg/L
  MK was produced using detergent supplement culture and a
  mutant of Flavabacterium.
 Lactic acid bacteria are reported to produce MK with the
  yield of 29–123 g/L MK-7, MK-8, MK-9 and MK-10.
 In fermented soybeans, Bacillus subtilis produces
  menaquinones, the major component being MK-7 and the
  minor one being MK-6.
 Sumi studied production of MKs by the fermentation of okara
  with seven different natto bacilli.
Summary

 Fermentative productions of vitamins have many
  advantages compared to conventional chemical
  synthesis processes.

 Different     methods             like        media
  optimization, mutation and screening, genetic
  engineering and biocatalyst conversion have been
  used for improvement of the production of vitamins.
vitamin by fermnetation

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vitamin by fermnetation

  • 1. Fermentative Production of Vitamins: Indian and Global Scenario Irfan Ahmad M. Tech. Pharm. Tech. (Biotech.) National Institute of Pharmaceutical Education and Research, S.A.S. Nagar
  • 2. CONTENTS 1. Introduction to Vitamins. 2. Industrial Production of Vitamins. 3. Vitamins Market Scenario. 4. Fermentative Production of Water Soluble Vitamins. i. Vitamin B₁₂ ii. Vitamin B₂ iii. Vitamin C iv. Vitamin H 5. Fermentative Production of Fat Soluble Vitamins. i. Vitamin E ii. Vitamin K 6. Summary.
  • 3. Vitamins Vitamins are defined as essential micronutrients that are required in trace quantity and cannot be synthesized by mammals but are essential for metabolism of all living organisms VITAMINS Fat Soluble Vitamins Water Soluble Vitamins Vitamin A Vitamins B Vitamin D Vitamin C Vitamin E Vitamin H Vitamin K Vitamin P, etc.
  • 4. Role of Vitamins 1 Nutritional and Physiologic Role . 2 Food/Feed Additives 3 Therapeutic Agents 4 Health Aids 5 Technical Aids
  • 5. Industrial Production Of Vitamins Extraction Chemical Fermentation
  • 6. Industrial Production of Vitamins Production Method World Production (tonne) Biotech. Chem. Extr. 1980 1990 2000 2010 Thiamine (B1) + 1700 4200 3700 Riboflavin (B2) + 2000 2400 4400 Niacin + + 8500 22000 2800 Pantothenic acid + + 5000 7000 Pyridoxine (B6) + 1600 2550 3800 Biotin + + 2.7 2.5 88 112 Folic acid + 100 400 534 Vitamin B12 + 2 10 25 175 Vitamin C + + 40000 60000 10700 Vitamin A + + 16800 2500 2700 Vitamin D + + 5000 Tocopherol (E) + + + 6800 22000 30000 Vitamin K + 500 Survase SA, Bajaj IB, Singhal RS: Biotechnological production of vitamins
  • 7. Microbial Production of Vitamins Vitamin Enzyme (Microorganism) Vitamin C 2,5-diketo-D-gluconic acid reductase (Corynebacterium sp.). ucu Biotin Fermentation (Serratia marcescens); Multiple enzyme system (Bacillus sphaericus). Riboflavin Fermentation (Eremothecium ashbyii, Ashbya gossypii, Bacillus sp.). Fermentation (Propionibacterium shermanii, Pseudomonas Vitamin B12 denitrificans) Vitamin E Freshwater microalgae, Euglena gracilis. Shimizu S: Vitamins and related compounds: microbial production
  • 8. Fermentative Production of Vitamins Advantages Disadvantages i. Efficient, i. Complex, ii. Less Energy Intensive, iii. Carried out at ambient ii. Require Careful Monitoring, temperature and pressure, iii. Limited Operations Region, iv. Produce Renewable iv. Susceptible to Substrate and byproducts (e.g., Biomass), Product Inhibition, v. Low Cost of Waste Disposal, v. High Cost of Downstream vi. Highly enantioselective and Processing, regioselective, vii. Don’t suffer from consumer vi. High Risk of Contamination, consciousness with regard to vii. Tendency to Elicit Allergic safety. Manifestations.
  • 9. Vitamin Market Scenario • Europe represents the largest regional market; the US constitutes the single largest market globally. • Asia-Pacific is likely to emerge as the fastest growing market, with a CAGR of about 4.0% over the analysis period. • Vitamin E represents the largest segment, owing to the extensive use of these vitamins in cosmetics, pharmaceuticals, and food end-use applications • The cosmetics industry, though relatively small in terms of the percentage share, is emerging as a key end-user with a CAGR of 4.6% over the analysis period. • The global vitamin market was $2.3 billion in 2000. However, the vitamin market is expected to reach US$3.2 Billion by 2017.
  • 10. Top Five Sources of US Vitamin Imports China Belgium Other Germany India Netherland The vitamin industry was once dominated by a core group of 3% 3% producers in the developed economies 7% but is now far more open and 13% competitive as an increasing number of manufacturers in emerging 57% economies such as China and India 17% weigh in.
  • 11. Hoffmann La Roche (Switzerland), BASF (Germany), ADM (USA), Hubei Guangji (China), Merck (Germany) and Takeda (Japan) are some of the global leaders in vitamin production.
  • 12. Sales Single vitamin In 2011, the Indian vitamin segment 0.3% grew 7.5 per cent in volumes (against Single 0.1 decline in the year 2010) and 10.3 Mineral per cent in value (against 5.4 per cent 9% Tonics growth in 2010).By the end of 12% 2014, the vitamins and minerals category will be worth INR 21,174.6m ($486.7m), with an expected CAGR of 3% between 2009 and 2014. Multivitamins 79% Five of the top 10 players are foreign multinational firms. Among Indian companies, the largest vitamin producers are Wockhardt, Raptakos Brett, Piramal Healthcare and Alkem.
  • 13. WATER SOLUBLE VITAMINS Vitamin B₁₂ Vitamin B₂ Vitamin C Vitamin H
  • 14. Vitamin B₁₂( Cyanocobalamin)  Cyanocobalamin is the industrially produced stable cobalamin form which is not found in nature.  It is obtained exclusively by fermentation process.  Merck began production of vitamin B₁₂ by Pseudomonas denitrificans in 1952 and have improved the efficiency of culture more than 30- fold relative to the performance of the original soil isolates by genetic manipulations and microbial screening.  Mutagenic treatments have resulted in improved activity. Sumi: Microbial Production of Vitamin B12
  • 17. Downstream Process of Vitamin B₁₂
  • 18. Vitamin B2 (Riboflavin) • Riboflavin has been produced commercially by chemical synthesis, by fermentation and by a combination of fermentation and chemical synthesis. • Recently, fermentation route has been widely used as it produces the vitamin in a single step, resulting in substantial cost savings. In contrast, chemical processes are multistage, and incur a lot of cost. • Most of the producers like BASF, Roche, ADM/Aventis, Hubei Guangji prefer fermentative production of riboflavin over chemical process.
  • 19. Vitamin B₂ • Although bacteria (Clostridium sp.) and yeasts (Candida sp.) are good producers, two closely related ascomycete fungi, Eremothecium ashbyii and Ashbya gossypii, are considered the best riboflavin producers. Ashbya gossypii produces 40000 times more vitamin than it needs for its own growth. • Recently, Nippon Roche, Japan, has developed and commercialized a single step fermentative riboflavin production using a recombinant Bacillus subtilis strain, which effectively produces riboflavin directly from glucose in fed-batch operation. • Improved strains for the production of riboflavin were constructed through metabolic engineering using recombinant DNA techniques in Corynebacterium ammoniagenes. • Sybesma et al. developed Lactococcus lactis strain using both direct mutagenesis and metabolic engineering for simultaneous overproduction of both folate and riboflavin.
  • 20. Vitamin C (Ascorbic Acid) • L-ascorbic acid finds its use mainly in food industry, being a vitamin as well as an antioxidant. • Majority of commercially manufactured L-ascorbic acid is synthesized via Reichstein process using D-glucose as a starting material • Approximately 50 % of synthetic ascorbic acid is used in vitamins supplements and pharmaceutical preparations. • Because of its antioxidant properties and its potential to stimulate collagen production, it is also widely used as an additive to cosmetics. • The current global market of L-ascorbic acid is in excess of US$ 585 million with an annual growth rate of 3 %.
  • 21. Vitamin C (Ascorbic Acid) . D-Glu D-Sorbitol Reichestein Process L-Sorbose L- Ascorbic Diacetone-L- 2-Keto-L- acid Sorbose Gluconic acid methyl ester
  • 22. Bacterial Fermentation of Vitamin C 2-Keto-D-gluconic acid Sorbitol Pathway Pathway
  • 23. Yeast Based Fermentation Process • Saccharomyces cerevisiae and Zygosaccharomyces bailiiaccumulate produce L-ascorbic acid intracellularly when incubated with L-galactose. • Over-expression of the D-arabinose dehydrogenase and D- arabinono-1,4-lactone oxidase in Saccharomyces cerevisiae enhances this ability significantly.
  • 24. Algae Based Fermentation Process • Skatrud and Huss described a method that involved initial growth of Chlorella pyrenoidosa ATCC53170 in a fermentor with a carbon source that is sufficient for the cells to grow to an intermediate density. At the depleted stage, additional carbon source was added sequentially or continuously to maintain the carbon source concentration below a predetermined level until the addition is terminated. This resulted in the production of 1.45 g/L of L-ascorbic acid. • Euglena gracilis Z. is one of the few microorganisms which simultaneously produce antioxidant vitamins such as carotene (71 mg/L), vitamin C (86.5 mg/L) and vitamin E (30.1 mg/L).
  • 25. Vitamin H (Biotin)  Biotin (vitamin H) is one of the most fascinating cofactors involved in central pathways in pro- and eukaryotic cell metabolism.  While humans and animals require several hundred micrograms of biotin per day, most microbes, plants and fungi appear to be able to synthesize the cofactor themselves.  Biotin is added to many food, feed and cosmetic products, creating a world market of 10–30 t/year.  Majority of the biotin sold is synthesized chemically via Goldberg and Sternbach syntheses.  The chemical synthesis is linked with a high environmental burden, much effort has been put into the development of biotin- overproducing microbes
  • 26. Biosynthesis of Biotin bioF gene bioA gene bioD gene The conversion of bioB gene dethiobiotin to biotin has not been resolved.
  • 27. Fermentative Production of Biotin • Ogata et al. screened microorganisms and demonstrated that the bacterium B. sphaericus can excrete significant quantities of biotin synthetic pathway intermediates from precursor, Pimelic acid. • Brown et al. studied the production of biotin by recombinant strains of E. coli. The strain used was E. coli C268 having a bio A genotype and plasmid pTG3410 with two expression cassettes . • Biotin production under limiting growth conditions by Agrobacterium/Rhizobium HK4 transformed with a modified Escherichia coli bio operon.
  • 28. FAT SOLUBLE VITAMINS Vitamin E Vitamin K
  • 29. Vitamin E  Most abundant among fat soluble vitamins and has the highest antioxidant activity in vivo.  In nature, only photosynthetic organisms are capable of producing α-tocopherol.  In humans, ∞-tocopherol is believed to play a major role in prevention of light induced pathologies of the skin, eyes and degenerative disorders such as atherosclerosis, cardiovascular diseases and cancer.  Industrial application of ∞-tocopherol includes its use in preservation of food, in cosmetics and sunscreens.
  • 30. Vitamin E Extraction The synthetic form is a mixture of eight stereoisomers Extraction from oils is not collectively known as all-rac- efficient, as these typically alpha-tocopherol. contain low levels of Vitamin E ∞-tocopherol when administered in equal amounts, the bioavailability of natural with respect to synthetic ∞-tocopherol is 2:1. Chemical Synthesis Chemical Synthesis
  • 31. Vitamin E Fermentative Production of Vitamin K₂  Microalgae Euglena gracilis Z and marine microalgae Dunaliella tertiolecta produce ∞-tocopherol in concentrations higher than conventional foods.  Carballo-Cardenas et al. studied ∞-tocopherol production with Dunaliella tertiolecta and Tetraselmis suecica, demonstrating that nutrient composition can be used as a tool to improve α-tocopherol productivity.
  • 32. Vitamin K₂ Vitamin K₁ . Vitamin K Vitamin K₂
  • 33. Vitamin K Fermentative Production of Vitamin K₂  Tani and Taguchi have reported that as much as 182 mg/L MK was produced using detergent supplement culture and a mutant of Flavabacterium.  Lactic acid bacteria are reported to produce MK with the yield of 29–123 g/L MK-7, MK-8, MK-9 and MK-10.  In fermented soybeans, Bacillus subtilis produces menaquinones, the major component being MK-7 and the minor one being MK-6.  Sumi studied production of MKs by the fermentation of okara with seven different natto bacilli.
  • 34. Summary  Fermentative productions of vitamins have many advantages compared to conventional chemical synthesis processes.  Different methods like media optimization, mutation and screening, genetic engineering and biocatalyst conversion have been used for improvement of the production of vitamins.

Editor's Notes

  1. Glucanobacteroxidans, Brevibacterium, Corynebacterium and Pseudomaonas