Abstract
Objectives
To characterize a novel ene-reductase from Meyerozyma guilliermondii and achieve the ene-reductase-mediated reduction of activated C=C bonds.
Results
The gene encoding an ene-reductase was cloned from M. guilliermondii. Sequence homology analysis showed that MgER shared the maximal amino acid sequence identity of 57 % with OYE2.6 from Scheffersomyces stipitis. MgER showed the highest specific activity at 30 °C and pH 7 (100 mM sodium phosphate buffer), and excellent stereoselectivities were achieved for the reduction of (R)-carvone and ketoisophorone. Under the reaction conditions (30 °C and pH 7.0), 150 mM (R)-carvone could be completely converted to (2R,5R)-dihydrocarvone within 22 h employing purified MgER as catalyst, resulting in a yield of 98.9 % and an optical purity of >99 % d.e.
Conclusion
MgER was characterized as a novel ene-reductase from yeast and showed great potential for the asymmetric reduction of activated C=C bonds of α,β-unsaturated compounds.
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Acknowledgments
This work was financially supported by the National Key Basic Research Development Program of China (“973” Program, No. 2012CB721003), the Natural Science Foundation of China (No. 21276084), Shanghai Natural Science Foundation (No. 15ZR1408600) and National Major Science and Technology Projects of China (No. 2012ZX09304009).
Supporting information
Supplementary Figure 1—Multiple sequence alignment of MgER with the other nine Old Yellow Enzymes. Substrate binding sites are indicated.
Supplementary Figure 2—Phylogenetic relationship of amino acid sequences of MgER to other Old Yellow Enyzmes with known function.
Supplementary Figure 3—GC-MS spectrum of (2R,5R)-dihydrocarvone prepared by MgER. GC-EI-MS m/z (M+ 152 for C10H16O) 137, 109, 95, 81, 67, 55.
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Zhang, B., Zheng, L., Lin, J. et al. Characterization of an ene-reductase from Meyerozyma guilliermondii for asymmetric bioreduction of α,β-unsaturated compounds. Biotechnol Lett 38, 1527–1534 (2016). https://doi.org/10.1007/s10529-016-2124-1
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DOI: https://doi.org/10.1007/s10529-016-2124-1