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Biocatalytic Production of Glucosamine from N-Acetylglucosamine by Diacetylchitobiose Deacetylase

  • Jiang, Zhu (Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University) ;
  • Lv, Xueqin (Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University) ;
  • Liu, Yanfeng (Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University) ;
  • Shin, Hyun-dong (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology) ;
  • Li, Jianghua (Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University) ;
  • Du, Guocheng (Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University) ;
  • Liu, Long (Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University)
  • Received : 2018.05.27
  • Accepted : 2018.07.18
  • Published : 2018.11.28

Abstract

Glucosamine (GlcN) is widely used in the nutraceutical and pharmaceutical industries. Currently, GlcN is mainly produced by traditional multistep chemical synthesis and acid hydrolysis, which can cause severe environmental pollution, require a long prodution period but a lower yield. The aim of this work was to develop a whole-cell biocatalytic process for the environment-friendly synthesis of glucosamine (GlcN) from N-acetylglucosamine (GlcNAc). We constructed a recombinant Escherichia coli and Bacillus subtilis strains as efficient whole-cell biocatalysts via expression of diacetylchitobiose deacetylase ($Dac_{ph}$) from Pyrococcus furiosus. Although both strains were biocatalytically active, the performance of B. subtilis was better. To enhance GlcN production, optimal reaction conditions were found: B. subtilis whole-cell biocatalyst 18.6 g/l, temperature $40^{\circ}C$, pH 7.5, GlcNAc concentration 50 g/l and reaction time 3 h. Under the above conditions, the maximal titer of GlcN was 35.3 g/l, the molar conversion ratio was 86.8% in 3-L bioreactor. This paper shows an efficient biotransformation process for the biotechnological production of GlcN in B. subtilis that is more environmentally friendly than the traditional multistep chemical synthesis approach. The biocatalytic process described here has the advantage of less environmental pollution and thus has great potential for large-scale production of GlcN in an environment-friendly manner.

Keywords

References

  1. Chen JK, Shen CR, Liu CL. 2010. N-acetylglucosamine: production and applications. Marine Drugs 8: 2493-2516. https://doi.org/10.3390/md8092493
  2. Nakamura H. 2011. Application of glucosamine on human disease-Osteoarthritis. Carbohydr. Polym. 84: 835-839. https://doi.org/10.1016/j.carbpol.2010.08.078
  3. Hungerford DS, Jones LC. 2003. Glucosamine and chondroitin sulfate are effective in the management of osteoarthritis. J. Arthroplasty. 18: 5-9. https://doi.org/10.1054/arth.2003.50067
  4. Towheed TE. 2003. Current status of glucosamine therapy in osteoarthritis. Arthritis Rheum. 49: 601-604. https://doi.org/10.1002/art.11183
  5. Sitanggang AB, Wu HS, Wang SS, Ho YC. 2010. Effect of pellet size and stimulating factor on the glucosamine production using Aspergillus sp. BCRC 31742. Bioresour. Technol. 101: 3595-3601. https://doi.org/10.1016/j.biortech.2009.12.084
  6. Zhang J, Liu L, Li J, Du G, Chen J. 2012. Enhanced glucosamine production by Aspergillus sp. BCRC 31742 based on the time-variant kinetics analysis of dissolved oxygen level. Bioresour. Technol. 111: 507-511. https://doi.org/10.1016/j.biortech.2012.02.063
  7. Chen X, Liu L, Li J, Du G, Chen J. 2012. Improved glucosamine and N-acetylglucosamine production by an engineered Escherichia coli via step-wise regulation of dissolved oxygen level. Bioresour. Technol. 110: 534-538. https://doi.org/10.1016/j.biortech.2011.12.015
  8. Deng MD, Severson DK, Grund AD, Wassink SL, Burlingame RP, Berry A, et al. 2005. Metabolic engineering of Escherichia coli for industrial production of glucosamine and N-acetylglucosamine. Metab. Eng. 7: 201-214. https://doi.org/10.1016/j.ymben.2005.02.001
  9. Liu Y, Liu L, Shin HD, Chen RR, Li J, Du G, et al. 2013. Pathway engineering of Bacillus subtilis for microbial production of N-acetylglucosamine. Metab. Eng. 19: 107-115. https://doi.org/10.1016/j.ymben.2013.07.002
  10. Mine S, Ikegami T, Kawasaki K, Nakamura T, Uegaki K. 2012. Expression, refolding, and purification of active diacetylchitobiose deacetylase from Pyrococcus horikoshii. Protein Expr. Purif. 84: 265-269. https://doi.org/10.1016/j.pep.2012.06.002
  11. Kang Z, Yang S, Du G, Chen J. 2014. Molecular engineering of secretory machinery components for high-level secretion of proteins in Bacillus species. J. Ind. Microbiol. Biotechnol. 41: 1599-1607. https://doi.org/10.1007/s10295-014-1506-4
  12. Tanaka T, Fukui T, Fujiwara S, Atomi H, Imanaka T. 2004. Concerted action of diacetylchitobiose deacetylase and exobeta- D-glucosaminidase in a novel chitinolytic pathway in the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. J. Biol. Chem. 279: 30021-30027. https://doi.org/10.1074/jbc.M314187200
  13. Mine S, Niiyama M, Hashimoto W, Ikegami T, Koma D, Ohmoto T, et al. 2014. Expression from engineered Escherichia coli chromosome and crystallographic study of archaeal N,N'-diacetylchitobiose deacetylase. FEBS J. 281: 2584-2596. https://doi.org/10.1111/febs.12805
  14. Nicolas P, Mader U, Dervyn E, Rochat T, Leduc A, Pigeonneau N, et al. 2012. Condition-dependent transcriptome reveals high-level regulatory architecture in Bacillus subtilis. Science 335: 1103-1106. https://doi.org/10.1126/science.1206848
  15. Promchai R, Promdonkoy B, Tanapongpipat S, Visessanguan W, Eurwilaichitr L, Luxananil P. 2016. A novel salt-inducible vector for efficient expression and secretion of heterologous proteins in Bacillus subtilis. J. Biotechnol. 222: 86-93. https://doi.org/10.1016/j.jbiotec.2016.02.019
  16. Bertram R , Rigali S , Wood N , L ulko A T, K uipers O P, Titgemeyer F. 2011. Regulon of the N-acetylglucosamine utilization regulator NagR in Bacillus subtilis. J. Bacteriol. 193: 3525-3536. https://doi.org/10.1128/JB.00264-11
  17. Vincent F, Yates D, Garman E, Davies GJ, Brannigan JA. 2004. The three-dimensional structure of the N-acetylglucosamine- 6-phosphate deacetylase, NagA, from Bacillus subtilis: a member of the urease superfamily. J. Biol. Chem. 279: 2809-2816. https://doi.org/10.1074/jbc.M310165200
  18. Song Y, Li J, Shin HD, Du G, Liu L, Chen J. 2015. One-step biosynthesis of alpha-ketoisocaproate from L-leucine by an Escherichia coli whole-cell biocatalyst expressing an L-amino acid deaminase from Proteus vulgaris. Sci. Rep. 5: 12614. https://doi.org/10.1038/srep12614
  19. Westers L, Westers H, Quax WJ. 2004. Bacillus subtilis as cell factory for pharmaceutical proteins: a biotechnological approach to optimize the host organism. Biochim. Biophys. Acta 1694: 299-310. https://doi.org/10.1016/j.bbamcr.2004.02.011
  20. Zhang XZ, Cui ZL, Hong Q, Li SP. 2005. High-level expression and secretion of methyl parathion hydrolase in Bacillus subtilis WB800. Appl. Environ. Microbiol. 71: 4101-4103. https://doi.org/10.1128/AEM.71.7.4101-4103.2005
  21. Mine S, Ikegami T, Kawasaki K, Nakamura T, Uegaki K. 2012. Expression, refolding, and purification of active diacetylchitobiose deacetylase from Pyrococcus horikoshii. Protein Express. Purification 84: 265-269. https://doi.org/10.1016/j.pep.2012.06.002
  22. Shi F, Li K, Huan X, Wang X. 2013. Expression of NAD(H) kinase and glucose-6-phosphate dehydrogenase improve NADPH supply and L-isoleucine biosynthesis in Corynebacterium glutamicum ssp. lactofermentum. Appl. Biochem. Biotechnol. 171: 504-521. https://doi.org/10.1007/s12010-013-0389-6
  23. Blombach B, Schreiner ME, Holatko J, Bartek T, Oldiges M, Eikmanns BJ. 2007. L-valine production with pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum. Appl. Environ. Microbiol. 73: 2079-2084. https://doi.org/10.1128/AEM.02826-06

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