Biocatalytic production of chiral epichlorohydrin in organic solvents

Choi, Won Jae; Lee, Eun Yeol; Yoon, Sung Jun; Yang, Seung-Taek; Choi, Cha Yong

doi:10.1016/s1389-1723(00)80022-5

Cited by 43 publications

(13 citation statements)

References 10 publications

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“…The e.e.% of (S)-ECH increases with pH from 3 to 8, and thereafter decreases from pH 8 to 9. These findings are in accordance with several earlier reports showing pH optima of 7.0 to 8.0 for partial purified EH from A. niger (Choi et al, 1999) and R. glutinis (Kim et al, 2004;Lee 2007), which also indicate that the EH from domestic duck liver is very sensitive to pH and lost a large degree of its activity when pH is lower than 5.0. EH has an optimum pH of about 8.0 at which their activity is maximal.…”

Section: Effects Of Ph On Relative Activity Of Ehsupporting

confidence: 93%

“…An enantio-selective EH from an actinomycetes source has been observed in a Nocardia H8, showing enantioselectivity in the hydrolysis of racemic ECH (Weijers and De Bont, 1991). Fungal epoxide hydrolysis has been described for Aspergillus niger (Choi et al, 1999) and Rhodotorula glutinis (Weijers, 1997;Kim et al, 2004;Lee, 2007). However, microbial cells include only a small amount of EH and often need to be induced to express this enzyme.…”

Section: Introductionmentioning

confidence: 99%

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Kinetic investigation on enantioselective hydrolytic resolution of epichlorohydrin by crude epoxide hydrolase from domestic duck liver

Ling¹,

Wang³

et al. 2011

Afr. J. Biotechnol.

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Enantiopure epichlorohydrin is a valuable epoxide intermediate for preparing optically active pharmaceuticals. In this study, a novel epoxide hydrolase prepared from domestic duck liver was used as biocatalyst for producing (S) -epichlorohydrin from its racemates. To characterize the biocatalytic profiles of crude epoxide hydrolase, some effect factors were investigated. The crude epoxide hydrolase has an apparent optimal pH of 8.0 and an optimal temperature of 35°C. Fe 2+ and Mn 2+ both enhanced the activity of epoxide hydrolase to different degrees. The results of kinetic study revealed that a significant inhibition phenomenon is observed in this reaction process. The apparent kinetic parameters (viz., V max and K m ) and apparent substrate inhibition parameter K is were calculated with linear fitting. The developed production process indicates that the novel epoxide hydrolase from domestic duck liver is a highly efficient biocatalyst for preparing enantiopure epichlorohydrin.

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Section: Effects Of Ph On Relative Activity Of Ehsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Kinetic investigation on enantioselective hydrolytic resolution of epichlorohydrin by crude epoxide hydrolase from domestic duck liver

Ling¹,

Wang³

et al. 2011

Afr. J. Biotechnol.

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“…Improvement of asymmetric hydrolysis by whole cells was achieved through optimization of the growth medium composition and fermentation mode. EH enzymes obtained from A. niger strains can catalyze hydrolysis of epichlorohydrin, cyclohexene oxide, styrene oxide, nitrostyrene oxides, pyridyloxiranes, and related compounds [13,15,16,20,23,30], and have been applied in various syntheses of key, enantiopure, building blocks [27,34]. Partial purification of EH from A. niger SQ-6 suggests that this enzyme shows some similarities with the EH identified in A. niger LCP521 [28], including substrate specificity, optimum pH and regioselectivity.…”

Section: Discussionmentioning

confidence: 96%

Enzymatic resolution of racemic phenyloxirane by a novel epoxide hydrolase from Aspergillus niger SQ-6 and its fed-batch fermentation

Liu

Qian

et al. 2005

J IND MICROBIOL BIOTECHNOL

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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.

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“…After 35 min of reaction, the ee of (R)-epichlorohydrin reached >99% with 42.7% yield. The ArEH was found to be the most enantioselective among several EH that have been reported for the preparation of chiral epichlorohydrin [19,25]. When the (S)-enantiomer was completely converted, the (R)-enantiomer of the epichlorohydrin started to be hydrolyzed at a much higher rate.…”

Section: Time Course Of the Resolution Of Racemic Epichlorohydrinmentioning

confidence: 90%

“…In the two-phase system used here, the substrate epichlorohydrin was mainly distributed in the organic phase, which reduced the inhibition and the spontaneous hydrolysis of epichlorohydrin. It has been reported that cyclohexane is an appropriate organic phase for the kinetic resolution of epichlorohydrin by EH [25]. Although other organic solvents such as n-hexane and n-heptane could be also used as the organic phase based on the high remaining activity of ArEH, the final yield of (R)-epichlorohydrin was lower than that in cyclohexane (data not shown).…”

Section: Concentration Enzymementioning

confidence: 93%

Biosynthesis of (R)‐epichlorohydrin at high substrate concentration by kinetic resolution of racemic epichlorohydrin with a recombinant epoxide hydrolase

Jin

Liu

et al. 2013

Engineering in Life Sciences

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Research ArticleBiosynthesis of (R)-epichlorohydrin at high substrate concentration by kinetic resolution of racemic epichlorohydrin with a recombinant epoxide hydrolaseThe substrate concentration and yield were shown to be very low in the production of (R)-epichlorohydrin by hydrolysis of racemic epichlorohydrin using epoxide hydrolases in previous studies. In this work, we synthesized an epoxide hydrolase gene from Agrobacterium radiobacter and expressed it in Escherichia coli by the PCR assembly method. The recombinant A. radiobacter epoxide hydrolase (ArEH) was applied in the preparation of (R)-epichlorohydrin and, a yield of 42.7% with ≥99% enantiomeric excess (ee) from 25.6 mM racemic epichlorohydrin was obtained. However, the ee of (R)-epichlorohydrin was not able to reach 99% due to substrate and product inhibition when the substrate concentration was over 320 mM. Inhibition studies revealed that (S)-3-chloro-1,2-propanediol displayed noncompetitive inhibition in the conversion of (S)-epichlorohydrin but non-significant inhibition for (R)-epichlorohydrin. Moreover, ArEH was successfully applied in the preparation of (R)-epichlorohydrin at high substrate concentration by eliminating the substrate inhibition. The substrate concentration increased to 448 mM by intermittent feeding of the substrate and to 512 mM by using a two-phase reaction system, with a high yield (>27%) and ee (>98%) of (R)-epichlorohydrin. This is the first report of high-yield production of (R)-epichlorohydrin at high substrate concentration, laying the foundations for its application on the industrial scale.

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Biocatalytic production of chiral epichlorohydrin in organic solvents

Cited by 43 publications

References 10 publications

Kinetic investigation on enantioselective hydrolytic resolution of epichlorohydrin by crude epoxide hydrolase from domestic duck liver

Kinetic investigation on enantioselective hydrolytic resolution of epichlorohydrin by crude epoxide hydrolase from domestic duck liver

Enzymatic resolution of racemic phenyloxirane by a novel epoxide hydrolase from Aspergillus niger SQ-6 and its fed-batch fermentation

Biosynthesis of (R)‐epichlorohydrin at high substrate concentration by kinetic resolution of racemic epichlorohydrin with a recombinant epoxide hydrolase

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