Bioprocess Strategies for Vitamin B12 Production by Microbial Fermentation and Its Market Applications
Abstract
:1. Historical Overview
2. Structure of Cobalamin Derivatives and Functions as Enzyme Cofactors
3. Biosynthesis of Vitamin B12: The Aerobic and Anaerobic Pathways
4. Microbial Production of Vitamin B12: Bioprocess Optimization for Cyanocobalamin Production
4.1. Microbial Production in Pseudomonas denitrificans
4.2. Microbial Production in Propionibacterium freudenreichii
5. Vitamin B12 Downstream Processing and Post-Modification Strategies
6. Patents—State of the Art
Patent Application Number (Reference) | Name | Microorganism/Strain | Innovation | Volumetric Production | Year | |
---|---|---|---|---|---|---|
Propionibacterium genus | US4544633A [43] (Expired) | Process for producing vitamin B12 by the fermentation technique, and vitamin B12-producing microorganism | P. freudenreichii (IFO 12424, IFO 12391, IFO 12426) | Creation of propionic-resistant strains (P. freudenreichii FERM-86 and FERM-87) for enhanced CNCbl production | 15 mg/L | 1983 |
US6492141B1 [107] (Expired) | Process for the production of vitamin B12 | P. freudenreichii CBS 929.97 | O2 effect in production during the anaerobic phase and a "fill and draw" strategy for enhanced production | 19 mg/L | 1999 | |
US6187761B1 [117] (Expired) | Production and use of compositions comprising high concentrations of vitamin B12 activity | P. freudenreichii subsp. shermanii and P. denitrificans | Method for producing vitamin B12 and making highly concentrated compositions | 10 mg/L | 1999 | |
US7427397B2 [108] (Expired) | Probiotic Propionibacterium | Propionibacterium jensenii 702 | Propionibacterium jensenii as a probiotic | 0.0012 mg/L | 2004 | |
EP2376644B1 [118] (Active) | Process for the preparation of a fermentation broth | Lactobacillus plantarum DSM 22,118 and P. freudenreichii DSM 22120 | Fermentation media optimization and co-culture for folate and vitamin B12 production | 1.07 mg/L | 2009 | |
CN206828509U [110] (Active) | A device for producing propionic acid and co-producing vitamin B12 by semi-continuous fermentation | P. freudenreichii | Simultaneous production of propionic acid and vitamin B12 in a semicontinuous fermentation with propionic acid separation | 20.12 mg/L | 2017 | |
US9938554 [109] (Active) | Co-cultivation of Propionibacterium and yeast. | P. freudenreichii (ATCC 6207) and yeast cells (DSM 28271) | Co-culture of Propionibacterium and propionic-resistant yeast to decrease the chemical oxygen load (COD) of spent media | 16 mg/L | 2018 | |
US20200149084A1 [83] (Active) | Sequential co-culturing method for producing a vitamin- and protein-rich food product | Basidiomycota and P. freudenreichii | Co-culture of Basidiomycota genus strains and vitamin B12-producing strains for in situ food fortification | 0.0014 mg/L 1 | 2020 | |
IN201827044769 A [111] (Active) | Continuous process for co-production of vitamin B12 and organic acids | P. freudenreichii (ATCC 13673) | Co-production of vitamin B12 and organic acids in a continuous fermentation with a single bioreactor | 76.13 mg/L | 2020 | |
WO21041759 A1 [119] (Active) | Modified Propionibacterium and methods of use | P. freudenreichii (P. UF 1) | Generation of a vitamin B12-overproducing strain by introducing a mutation that decreases the activity of the cbiMcbl riboswitch | n.d. 2 | 2021 | |
Pseudomonas denitrificans | US3018225A [100] (Expired) | Production of vitamin B12 | P. denitrificans MB-580 | A process for vitamin B12 production with a high-yield strain (P. denitrificans MB-580) | 2.4 mg/L 1 | 1962 |
US20060019352A1 [101] (Abandoned) | Methods for increasing the production of cobalamins using cob gene expression | P. denitrificans | Overexpression of several genes involved in Cob biosynthesis; generation of several overproducing strains, such as SC-510 | 65 mg/L | 1990 | |
US6156545A [120] (Expired) | Biosynthesis method enabling the preparation of cobalamins | P. denitrificans G2650 | Enhanced Cob production by the heterologous overexpression of precursors, such as DMBI and O-phospo-L-threonine | 7.9 mg/L | 1996 | |
CN101538599A [121] (Active) | Method for improving the yield of denitrified pseudomonas vitamin B12 | P. denitrificans J741 | Enhance cob production by betaine addition optimization | 177.49 mg/L | 2008 | |
CN102399845A [122] (Active) | Vitamin B12 fermentation production control process based on CO2 concentration in tail gas | P. denitrificans MB-580 | Vitamin B12 enhanced production through a carbon dioxide control strategy during fermentation | 164.6 mg/L | 2010 | |
CN101748177 A [123] (Active) | Optimized method for producing vitamin B12 through P. denitrificans fermentation and synthetic medium | P. denitrificans | Development and optimization of media and bioprocess conditions for improved vitamin B12 production | 77 mg/L | 2010 | |
CN102021214 A [124] (Active) | Oxygen consumption rate-based vitamin B12 fermentation production control process | P. denitrificans | Vitamin B12 production optimization through an oxygen control strategy | 171,4 mg/L | 2011 | |
CN102453740 A [125] (Active) | Culture medium for producing vitamin B12 by fermenting P. denitrificans and fermentation method thereof | P. denitrificans | Use of artificial molasses and bioprocess optimization for a more stable fermentation yield | 198 mg/L | 2012 | |
CN108949866 A [103] (Active) | Multi-stage rotating speed regulating policy for improving P. denitrificans fermentation for production of vitamin B12 | P. denitrificans | Vitamin B12 production improved by optimization of the culture media and the stirring speed of the bioprocess | 246 mg/L 1 | 2018 | |
CN108913739 A [126] (Active) | Method for producing vitamin B12 by using P. denitrificans based on pH value control | P. denitrificans | Improved vitamin B12 production by optimization of the bioprocess through pH value control | 248 mg/L | 2018 | |
CN110205350 A [104] (Active) | Method for improving the yield of vitamin B12 based on the regulation of ammonia nitrogen index | P. denitrificans | A method for improved Cbl production by supplementation with yeast extract controlled by the ammonia nitrogen index | 167 mg/L 1 | 2019 | |
CN109837320 A [105] (Active) | Method for promoting P. denitrificans to generate vitamin B12 | P. denitrificans | Optimization of media and culture conditions for improved vitamin B12 production | 198 mg/L | 2019 | |
CN111808158 A [106] (Active) | Preparation method of vitamin B12 crude product | P. denitrificans | Downstream process improvement for AdoCbl extraction | n.d. 2 | 2020 | |
CN111254173 A [102] (Active) | Screening method and screening culture medium for bacterial strains for high yield of vitamin B12 produced through fermentation production with P. denitrificans | Several high-yield strains of P. denitrificans | Screening for high-vitamin B12 producing P. denitrificans strains and culture medium screening for high vitamin B12 production | 281 mg/L 1 | 2020 | |
Other producers | US2650896A [127] (Expired) | Cyanide ions in production of vitamin B12 | Streptomyces griseus | Effects of cyanide ions in B12 production | Biological assay | 1953 |
US2576932A [112] (expired) | Fermentation process to produce vitamin B12 | B. megaterium B-938 | Vitamin B12 production with B. megaterium in a nutrient media with sucrose | 0.45 mg/L | 1983 | |
US20050227332A1 [128] (Expired) | Method for producing vitamin B12 from hydrogen-metabolizing methane bacterium | A mesophilic methane bacterium obtained from digested sludge | The culture is acclimatized in a H2–CO media and grown in an immobilized bed bioreactor | 25.2 mg/L | 2005 | |
US20060105432A1 [129] (Abandoned) | Method for the production of vitamin B12 | B. megaterium DSMZ509 | Genetically modified B. megaterium strain | 0.008 mg/L 1 | 2006 | |
WO2011154820A2 [113] (Application granted) | Vitamin B12-producing probiotic bacterial strains | Lactobacillus reuteri (DSM 17938, DSM 16143, ATCC 55730) | In situ food fortification for increased vitamin B12 production with Lactobacillus reuteri strains | 0.018 mg/L 1 | 2011 | |
CN104342390 A [114] (Active) | Sinorhizobium meliloti strain and composition and application of Sinorhizobium meliloti strain | S. meliloti (CGMCC 9638) | A S. melitolli strain capable of producing vitamin B12 and optimization of the bioprocess for vitamin B12 production | At least 50 mg/L | 2015 | |
WO2019109975A1 [116] (Active) | Recombinant strain of Escherichia coli for de novo synthesis of vitamin B12, construction method therefor and application thereof | E. coli | Recombinant E. coli for the de novo synthesis of vitamin B12 | 89 µg/g DCW | 2019 | |
CN110804598 A [115] (Active) | Procorrin-2C(20)-methyltransferase mutant and mutant gene and application thereof in preparing vitamin B12 | Sinorhizobium (CGMCC 9638) | Generation of a vitamin B12 overproducer strain by overexpressing the precorrin-2C(20)-methyltransferase gene | 115 mg/L | 2020 |
7. Vitamin B12 Market Applications and the State of the Market
8. Concluding Remarks
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Microorganism/Strain | Main Media Components | Scale | Summary/Innovation | Volumetric Production | Volumetric Productivity (mg/L/h) | Reference |
---|---|---|---|---|---|---|
B. megaterium DSM 319 | Terrific broth media | 250 mL shake flask | Precursor supplementation and pO2 control | 0.21 mg/L c | 0.006 mg/L/h c | [47] |
Lactobacillus reuteri ZJ03 | Soymilk | 250 mL shake flask | Different carbon source supplementation | 0.204 mg/L | 0.003 mg/L/h | [49] |
P. freudenreichii subsp. shermanii NRRL-B-4327, 3523 and NRRL-B-3524 | Sodium lactate broth | 250 mL shake flask | Vitamin B12 analogue addition | 31 mg/L | 0.51 mg/L | [50] |
P. freudenreichii CICC 10019 | Glucose, CSL a | 7 L stirred tankbioreactor | Expanded-bed bioreactor (EBAB) with crop stark hydrolysates | 47.6 mg/L | 0.18 mg/L/h | [51] |
P. freudenreichii CICC 10019 | Glucose, CSL | 7 L stirred tank bioreactor | EBAB bioreactor | 43.4 mg/L | 0.27 mg/L/h | [52] |
P. freudenreichii CICC 10019 | Glucose, CSL | 1.5 L stirred tank bioreactor | EBAB bioreactor and DMBI addition | 58.8 mg/L | 0.59 mg/L/h | [53] |
P. freudenreichii CICC 10019 | Glucose/glycerol, CSL | 5 L stirred tank bioreactor | EBAB bioreactor, glycerol as carbon source and crop stalk hydrolysate as nitrogen source | 43 mg/L | 0.36 mg/L/h | [54] |
P. freudenreichii DF13 | Supplemented whey permeate | 1 L stirred tank bioreactor | Co-culture with Lactobacillus plantarum SM39 for simultaneous folate and Cbl production | 0.75 mg/L | 0.004 mg/L/h | [55] |
P. freudenreichii DSM 20271//Lactobacillus brevis ATCC 14869 | Wheat bran dough | n.d. b | Co-fermentation in wheat bran dough for in situ production of Vitamin B12 | 332 ng/g c | n.d. b | [56] |
P. freudenreichii IFO 12424//Ralstonia eutropha H16 (ATCC17699) | Polypeptone, casein, yeast extract | 5 L stirred tank bioreactor | Cell recycling system and co-culture with Ralstonia eutropha for decreasing propionic acid inhibition | 8 mg/L c | 0.14 mg/L/h c | [57] |
P. freudenreichii PTCC 1674. | Tryptone, yeast extract, different carbon sources | 100 cm3 | Waste frying sun oil as a carbon source for vitamin B12 production | 2.74 mg/L | 0.02 mg/L/h | [58] |
P. freudenreichii subsp. shermanii ATCC 13673 | Glucose, yeast extract | 2 L stirred tank bioreactor | Inoculum volume, pH control and substrate concentration optimization | 0.087 mg/L | 0.002 mg/L/h | [59] |
P. freudenreichii subsp. shermanii CICC 10019 | Glucose, CSL | 100 L fermenter | Addition of DMBI precisely with Ado-Cbl control strategy | 39.15 mg/L | 0.32 mg/L/h | [60] |
P. freudenreichii subsp. shermanii | Glycerol, tryptone, casein, DMBI | 200 mL shake flask | Media optimization by design of experiments with crude glycerol as the main carbon source | 4.01 mg/L | 0.024 mg/L/h | [61] |
P. freudenreichii subsp. shermanii | Whey based media | 20 mL tubes | DMBI, Nicotinamide and Riboflavin supplementation | 5.3 mg/L | 0.03 mg/L/h | [62] |
P. freudenreichii subsp. shermanii | Food-like media (cereal matrices) | n.d. b | Precursor supplementation in different cereal-like matrices | 1.5 mg/Kg | 0.009 mg/Kg/h | [63] |
P. freudenreichii subsp. shermanii 2067 | Cheese-based propionic media/whey-based liquid media | 50 mL shake flask | Production in food-like conditions without DMBI addition | 0.124 mg/L c | 0.0013 mg/L/h | [64] |
P. freudenreichii CICC10019 | Glucose, yeast extract, CSL | 100 mL flasks | Media optimization by statistical analysis | 8.32 mg/L | 0.068 mg/L/h | [65] |
P. freudenreichii CICC10019 | Glucose, CSL | 7 L fermenter | Membrane separation-coupled fed-batch fermentation | 21.6 mg/L | 0.16 mg/L/h | [66] |
P. denitrificans | Maltose, peptone, betaine | 250 mL shake flask | Addition of rotenone as a respiration inhibitor for enhanced production | 54.7 mg/L | 0.57 mg/L/h | [67] |
P. denitrificans | Beet molasses, sucrose, betaine | 120 m3 fermenter | Glucose-betaine feeding, pH control strategy | 214.13 mg/L c | 1.27 mg/L/h c | [68] |
P. denitrificans | Glucose, CSL, betaine | 120 m3 fermenter | Stepwise oxygen uptake rate control strategy | 188 mg/L | 1.12 mg/L/h | [69] |
P. denitrificans | Glucose, CSL, betaine | 50 L fermenter | Effects of specific oxygen consumption rate on cell morphology and production | 213.1 mg/L | 1.88 mg/L/h | [70] |
P. denitrificans | Maltose, peptone, betaine | 250 mL shake flask | Betaine supplementation | 58.61 mg/L | 0.48 mg/L/h | [71] |
P. denitrificans | Maltose syrup, CSL, betaine | 120 m3 fermenter | Maltose syrup and CSL as the main substrates | 198.27 mg/L | 1.10 mg/L/h | [72] |
P. denitrificans | Glucose, CSL, betaine | 120 m3 fermenter | pO2 stepwise control | 198.80 mg/L | 1.18 mg/L/h | [73] |
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Calvillo, Á.; Pellicer, T.; Carnicer, M.; Planas, A. Bioprocess Strategies for Vitamin B12 Production by Microbial Fermentation and Its Market Applications. Bioengineering 2022, 9, 365. https://doi.org/10.3390/bioengineering9080365
Calvillo Á, Pellicer T, Carnicer M, Planas A. Bioprocess Strategies for Vitamin B12 Production by Microbial Fermentation and Its Market Applications. Bioengineering. 2022; 9(8):365. https://doi.org/10.3390/bioengineering9080365
Chicago/Turabian StyleCalvillo, Álvaro, Teresa Pellicer, Marc Carnicer, and Antoni Planas. 2022. "Bioprocess Strategies for Vitamin B12 Production by Microbial Fermentation and Its Market Applications" Bioengineering 9, no. 8: 365. https://doi.org/10.3390/bioengineering9080365