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Industrial vitamin B12 production by Pseudomonas denitrificans using maltose syrup and corn steep liquor as the cost-effective fermentation substrates

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Abstract

The aerobic Pseudomonas denitrificans is widely used for industrial and commercial vitamin B12 fermentation, due to its higher productivity compared to the anaerobic vitamin B12-producing microorganisms. This paper aimed to develop a cost-effective fermentation medium for industrial vitamin B12 production by P. denitrificans in 120,000-l fermenter. It was found that maltose syrup (a low-cost syrup from corn starch by means of enzymatic or acid hydrolysis) and corn steep liquor (CSL, a by-product of starch industry) were greatly applicable to vitamin B12 production by P. denitrificans. Under the optimal fermentation medium performed by response surface methodology, 198.27 ± 4.60 mg/l of vitamin B12 yield was obtained in 120,000-l fermenter, which was close to the fermentation with the refined sucrose (198.80 mg/l) and was obviously higher than that obtained under beet molasses utilization (181.75 mg/l). Therefore, maltose syrups and CSL were the efficient and economical substrates for industrial vitamin B12 fermentation by P. denitrificans.

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References

  1. Martins JH, Barg H, Warren MJ, Jahn D (2002) Microbial production of vitamin B12. Appl Microbiol Biotechnol 58:275–285

    Article  Google Scholar 

  2. Randaccio L, Geremia S, Demitri N, Wuerges J (2010) Vitamin B12: unique metalorganic compounds and the most complex vitamins. Molecules 15:3228–3259

    Article  CAS  Google Scholar 

  3. Sally P, Stabler MD (2014) Vitamin B12 deficiency. New Engl J Med 368:149–160

    Google Scholar 

  4. Takahashi-Iñiguez T, García-Hernandez E, Arreguín-Espinosa R, Flores ME (2012) Role of vitamin B12 on methylmalonyl-CoA mutase activity. Zhejiang Univ-Sci B (Biomed Biotechnol) 13(6):423–437

    Article  Google Scholar 

  5. Survase SA, Bajaj IB, Singhal RS (2006) Biotechnological production of vitamins. Food Technol Biotechnol 44(3):381–396

    CAS  Google Scholar 

  6. Kang Z, Zhang J, Zhou J, Qi QS, Du GC, Chen J (2012) Recent advances in microbial production of δ-aminolevulinic acid and vitamin B12. Biotechnol Adv 30(6):1533–1542

    Article  CAS  Google Scholar 

  7. Wang P, Wang YS, Su ZG (2012) Improvement of adenosylcobalamin production by metabolic control strategy in Propionibacterium freudenreichii. Appl Biochem Biotechnol 167:62–72

    Article  CAS  Google Scholar 

  8. Li KT, Zhou J, Cheng X, Wei SJ (2012) Study on the dissolved oxygen control strategy in large-scale vitamin B12 fermentation by Pseudomonas denitrificans. J Chem Technol Biotechnol 87:1648–1653

    Article  CAS  Google Scholar 

  9. Li KT, Liu DH, Chu J, Wang YH, Zhuang YP, Zhang SL (2008) An effective and simplified pH-stat control strategy for the industrial fermentation of vitamin B12 by Pseudomonas denitrificans. Bioproc Biosyst Eng 31:605–610

    Article  CAS  Google Scholar 

  10. Li KT, Liu DH, Li YL, Chu J, Wang YH, Zhuang YP, Zhang SL (2008) Improved large-scale production of vitamin B12 by Pseudomonas denitrificans with betaine feeding. Bioresour Technol 99:8516–8520

    Article  CAS  Google Scholar 

  11. Li KT, Peng WF, Zhou J, Wei SJ, Cheng X (2013) Establishment of beet molasses as the fermentation substrate for industrial vitamin B12 production by Pseudomonas denitrificans. J Chem Technol Biotechnol 88:1730–1735

    Article  CAS  Google Scholar 

  12. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  13. Niu XH, Yue ZH, Zhang QM (2007) HPLC-ELED determination of glucose, maltotriose and other related substances in maltose. Chin J Pharm Anal 27(8):1261–1263

    CAS  Google Scholar 

  14. Prabakaran G, Hoti SL (2008) Influence of amino nitrogen in the culture medium enhances the production of δ-endotoxin and biomass of Bacillus thuringiensis var. israelensis for the large-scale production of the mosquito control agent. J Ind Microbiol Biotechnol 35:961–965

    Article  CAS  Google Scholar 

  15. Ebert RF (1986) Aminoacid analysis by HPLC: optimized conditions for chromatography of phenylthiocarbamyl derivatives. Anal Biochem 154(2):431–435

    Article  CAS  Google Scholar 

  16. Murthy GS, Johnston DB, Rausch KD, Tumbleson ME, Singh V (2011) Starch hydrolysis modeling: application to fuel ethanol production. Bioprocess Biosyst Eng 34:879–890

    Article  CAS  Google Scholar 

  17. Khan I, Nazir K, Wang ZP, Liu GL, Chi ZM (2014) Calcium malate overproduction by Penicillium viticola 152 using the medium containing corn steep liquor. Appl Microbiol Biotechnol 98:1539–1546

    Article  CAS  Google Scholar 

  18. Sharma N, Prasad GS, Choudhury AR (2013) Utilization of corn steep liquor for biosynthesis of pullulan, an important exopolysaccharide. Carboh Polym 93:93–101

    Article  Google Scholar 

  19. Warren MJ, Raux E, Schubert HL, Escalante-Semerena JC (2002) The biosynthesis of adenosylcobalamin (vitamin B12). Nat Prod Rep 19:390–412

    Article  CAS  Google Scholar 

  20. Demain AL, Daniels HJ, Schnabel L, White RF (1968) Specificity of the stimulatory effect of betaine on the vitamin B12 fermentation. Nature 220:1324–1325

    Article  CAS  Google Scholar 

  21. Demain AL, White RF (1971) Porphyrin overproduction by Pseudomonas denitrificans: essentiality of betaine and stimulation by ethionine. J Bacteriol 107:456–460

    CAS  Google Scholar 

  22. Roman RV, Iluc E, Mustea A, Neacsu A, Asandului V (2001) Optimization of medium components in vitamin B12 biosynthesis. Roum Biotechnol Lett 6:343–350

    Google Scholar 

  23. Arockiasamy S, Krishnan IPG, Anandakrishnan N, Seenivasan S, Sambath A, Venkatasubramani JP (2008) Enhanced production of laccase from Coriolus versicolor NCIM 996 by nutrient optimization using using response surface methodology. Appl Biochem Biotech 151(2–3):371–379

    Article  CAS  Google Scholar 

  24. Singh N, Rai V, Tripathi CKM (2012) Production and optimization of oxytetracycline by a new isolate Streptomyces rimosus using response surface methodology. Med Chem Res 21(10):3140–3145

    Article  CAS  Google Scholar 

  25. Wang ZJ, Wang HY, Li YL, Chu J, Huang MZ, Zhuang YP, Zhang SL (2010) Improved vitamin B12 production by step-wise reduction of oxygen uptake rate under dissolved oxygen limiting level during fermentation process. Bioresour Technol 101(8):2845–2852

    Article  CAS  Google Scholar 

  26. van Hoek P, Aristidou A, Hahn JJ, Patist A (2003) Fermentation goes large scale. Chem Eng Prog 99:37–42

    Google Scholar 

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (Grant No. 31460019), Jiangxi Key Project of Basic Research Program (20143ACB21005), Training Program for Young Scientists of Jiangxi Provincial Department of Science and Technology (20142BCB23025), and International Scientific and Technological Cooperation Projects of Jiangxi Provincial Department of Science and Technology (20141BDH80033). We are also grateful to Shijiazhuang Pharma Group Hua Rong Pharmaceutical Co. Ltd for supporting this work.

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  1. Nanchang Key Laboratory of Applied Fermentation Technology, Jiangxi Agricultural University, Nanchang, 330045, China

    Wei Xia, Wei Chen, Wei-fu Peng & Kun-tai Li

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  1. Wei Xia
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  2. Wei Chen
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  3. Wei-fu Peng
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  4. Kun-tai Li
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Correspondence to Kun-tai Li.

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Xia, W., Chen, W., Peng, Wf. et al. Industrial vitamin B12 production by Pseudomonas denitrificans using maltose syrup and corn steep liquor as the cost-effective fermentation substrates. Bioprocess Biosyst Eng 38, 1065–1073 (2015). https://doi.org/10.1007/s00449-014-1348-5

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