Abstract
Objectives
To investigate the roles of adhS, which encodes the AdhS subunit of membrane-bound alcohol dehydrogenase (mADH) in Gluconobacter oxydans DSM2003, and to rationally improve mADH activity.
Results
adhS was identified and overexpressed in G. oxydans DSM2003. Its overexpression promoted the AdhA subunit which serves as the primary dehydrogenase transfer from the periplasmic space to the periplasmic surface of the membrane thereby increasing the amount of active mADH and thus enhancing mADH activity up to 1.96-fold. The increased mADH activity significantly altered product selectivity (glyceric acid/dihydroxyacetone) during glycerol oxidation and increased the glyceric acid production by 7.6-fold. By comparison, overexpression of adhS and adhABS was equally effective in increasing the mADH activity and glyceric acid production.
Conclusions
adhS overexpression effectively improved mADH activity, indicating that for mADH, adhS might be a limiting component. The findings provide a guide for the efficient application of Gluconobacter spp. in hydroxy acid production.
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Acknowledgments
This work was supported by the National Key Basic Research Development Program of China (“973” Program; No. 2012CB721003), the Shanghai Natural Science Foundation (No. 15ZR1408600), the National Major Science and Technology Projects of China (No.2012ZX09304009), and the National High Technology Research and Development Program of China (“863” Program; No. 2012AA022201C).
Supporting Information
Supplementary Table 1—Primers used in this study.
Supplementary Figure 1—Nucleotide and amino acid sequences of adhS and upstream promoter in Gluconobacter oxydans DSM 2003.
Supplementary Figure 2—Comparison of amino acid sequences deduced from adhS in four Gluconobacter species.
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Zhang, H., Shi, L., Lin, J. et al. Effective improvement of the activity of membrane-bound alcohol dehydrogenase by overexpression of adhS in Gluconobacter oxydans . Biotechnol Lett 38, 1131–1138 (2016). https://doi.org/10.1007/s10529-016-2084-5
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DOI: https://doi.org/10.1007/s10529-016-2084-5