Publication: Kinetic properties and stability of glucose dehydrogenase from Bacillus amyloliquefaciens SB5 and its potential for cofactor regeneration
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Issued Date
2015
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eng
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Mahidol University
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springerOpen Journal
Bibliographic Citation
. AMB Expr. Vol.5, (2015), 68
Suggested Citation
Thunyarat Pongtharangkul, Pattra Chuekitkumchorn, Nhuengtida Suwanampa, Panwajee Payongsri, Kohsuke Honda, Watanalai Panbangred Kinetic properties and stability of glucose dehydrogenase from Bacillus amyloliquefaciens SB5 and its potential for cofactor regeneration. . AMB Expr. Vol.5, (2015), 68. doi:10.1186/s13568-015-0157-9 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/3025
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Title
Kinetic properties and stability of glucose dehydrogenase from Bacillus amyloliquefaciens SB5 and its potential for cofactor regeneration
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Abstract
Glucose dehydrogenases (GluDH) from Bacillus species offer several advantages over other NAD(P)H regeneration
systems including high stability, inexpensive substrate, thermodynamically favorable reaction and flexibility to regenerate
both NADH and NADPH. In this research, characteristics of GluDH from Bacillus amyloliquefaciens SB5 (GluDHBA)
was reported for the first time. Despite a highly similar amino acid sequence when comparing with GluDH from
Bacillus subtilis (GluDH-BS), GluDH-BA exhibited significantly higher specific activity (4.7-fold) and stability when pH
was higher than 6. While an optimum activity of GluDH-BA was observed at a temperature of 50 °C, the enzyme was
stable only up to 42 °C. GluDH-BA exhibited an extreme tolerance towards n-hexane and its respective alcohols. The
productivity of GluDH obtained in this study (8.42 mg-GluDH/g-wet cells; 1035 U/g-wet cells) was among the highest
productivity reported for recombinant E. coli. With its low KM-value towards glucose (5.5 mM) and NADP+ (0.05 mM),
GluDH-BA was highly suitable for in vivo applications. In this work, a recombinant solvent-tolerant B. subtilis BA overexpressing
GluDH-BA was developed and evaluated by coupling with B. subtilis overexpressing an enzyme P450 BM3
F87V for a whole-cell hydroxylation of n-hexane. Significantly higher products obtained clearly proved that B. subtilis
BA was an effective cofactor regenerator, a valuable asset for bioproduction of value-added chemicals.