Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 102, Issue 13, Pages 5545-5556Publisher
SPRINGER
DOI: 10.1007/s00253-018-8984-7
Keywords
Rhodococcus; Ene-reductase; Enantioselectivity; Asymmetric reduction
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Funding
- China Scholarship Council-Delft University of Technology Joint Program
- Netherlands Organization for Scientific Research (NWO)
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Rhodococcus strains are ubiquitous in nature and known to metabolise a wide variety of compounds. At the same time, asymmetric reduction of C=C bonds is important in the production of high-valued chiral building blocks. In order to evaluate if Rhodococci can be used for this task, we have probed several Rhodococcus rhodochrous and R. erythropolis strains for ene-reductase activity. A series of substrates including activated ketones, an aldehyde, an imide and nitro-compound were screened using whole cells of seven Rhodococcus strains. This revealed that whole cells of all Rhodococcus strains showed apparent (S)-selectivity towards ketoisophorone, while most other organisms show (R)-selectivity for this compound. Three putative ene-reductases from R. rhodochrous ATCC 17895 were heterologously expressed in Escherichia coli. One protein was purified and its biocatalytic and biochemical properties were characterised, showing typical (enantioselective) properties for class 3 ene-reductases of the old yellow enzyme family.
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