Enzymatic resolution of racemic phenyloxirane by a novel epoxide hydrolase from Aspergillus niger SQ-6 and its fed-batch fermentation.
A microorganism with the ability to catalyze the resolution of racemic phenyloxirane was isolated and identified as Aspergillus niger SQ-6. Chiral capillary electrophoresis was successfully applied to separate both phenyloxirane and phenylethanediol. The epoxide hydrolase (EH) involved in this resolution process was (R)-stereospecific and constitutively expressed. When whole cells were used during the biotransformation process, the optimum temperature and pH for stereospecific vicinal diol production were 35 degrees C and 7.0, respectively. After a 24-h conversion, the enantiomer excess of (R)-phenylethanediol produced was found to be >99%, with a conversion rate of 56%. In fed-batch fermentations at 30 degrees C for 44 h, glycerol (20 g L(-1)) and corn steep liquor (CSL) (30 g L(-1)) were chosen as the best initial carbon and nitrogen sources, and EH production was markedly improved by pulsed feeding of sucrose (2 g L(-1) h(-1)) and continuous feeding of CSL (1 g L(-1) h(-1)) at a fermentation time of 28 h. After optimization, the maximum dry cell weight achieved was 24.5+/-0.8 g L(-1); maximum EH production was 351.2+/-13.1 U L(-1) with a specific activity of 14.3+/-0.5 U g(-1). Partially purified EH exhibited a temperature optimum at 37 degrees C and pH optimum at 7.5 in 0.1 M phosphate buffer. This study presents the first evidence for the existence of a predicted epoxide racemase, which might be important in the synthesis of epoxide intermediates.
Liu
Yanbin
Y
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beiyitiao 13, Zhongguancun, Haidian, Beijing, 100080, People's Republic of China.
Sha
Qian
Q
Wu
Sheng
S
Wang
Jianjun
J
Yang
Liu
L
Sun
Wanru
W
D001234:Aspergillus niger / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism*; D019075:Electrophoresis, Capillary; D004851:Epoxide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D004852:Epoxy Compounds / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D005026:Ethylene Glycols / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D013237:Stereoisomerism; D013696:Temperature
D004852:Epoxy Compounds; D005026:Ethylene Glycols; C031474:styrene glycol; C013690:styrene oxide; D004851:Epoxide Hydrolases