Rhodococcus strains as source for ene-reductase activity
Bi Shuang Chen (TU Delft - BT/Biocatalysis, Sun Yat-sen University)
Rosario Médici (TU Delft - BT/Biocatalysis)
Michelle P. van der Helm (TU Delft - BT/Biocatalysis)
Ymke van Zwet (TU Delft - BT/Biocatalysis)
Lorina Gjonaj (Leiden University Medical Center, TU Delft - BT/Biocatalysis)
Roelien van der Geest (TU Delft - BT/Biocatalysis)
Linda G. Otten (TU Delft - BT/Biocatalysis)
Ulf Hanefeld (TU Delft - BT/Biocatalysis)
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Abstract
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.