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Efficient production of ethyl (R)-2-hydroxy-4-phenylbutyrate using a cost-effective reductase expressed in Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
3113 151 2014 6 PDF Available
Title
Efficient production of ethyl (R)-2-hydroxy-4-phenylbutyrate using a cost-effective reductase expressed in Pichia pastoris
Abstract

•CgKR2 was successfully expressed in Pichia pastoris with high level.•Up to 1 M OPBE was converted to (R)-HPBE using CgKR2 as catalyst.•Secreted CgKR2 showed to be a better catalyst.•CgKR2 is cost-effective.

Recombinant strains of Pichia pastoris expressing carbonyl reductase CgKR2 from Candida glabrata were constructed for stereoselective reduction of ethyl-2-oxo-4-phenylbutyrate (OPBE) to ethyl (R)-2-hydroxy-4-phenylbutyrate [(R)-HPBE], an important building block for synthesis of angiotensin-converting enzyme (ACE) inhibitors. An intracellular expression level of 6.67 g L−1 and a secretion expression level of 3.0 g L−1 were obtained, respectively, by high cell density fermentation. By using whole cells of KM71/CgKR2 in aqueous system, the CgKR2-mediated bioreduction was performed, achieving a complete conversion of 1.0 M OPBE at 0.5 L scale, with a final yield of 77.9% and an enantiomeric excess (ee) of 97.3%. The secreted enzyme CgKR2 appeared to be a better catalyst with respect to the perfect ee of the product (R)-HPBE (>99%) when comparing with the recombinant cells of KM71/CgKR2. The cost of the resultant (R)-HPBE was reduced by 1/4 using the secreted CgKR2.

Keywords
Biocatalysis; (R)-HPBE; Enzyme production; Recombinant Pichia pastoris; Fermentation; Cost-effective reductase; Fed-batch culture
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Efficient production of ethyl (R)-2-hydroxy-4-phenylbutyrate using a cost-effective reductase expressed in Pichia pastoris
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 91, 15 October 2014, Pages 72–77
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us