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Co-immobilised aspartase and transaminase for high-yield synthesis of l-phenylalanine

Paper ID Volume ID Publish Year Pages File Format Full-Text
3069 149 2015 6 PDF Available
Title
Co-immobilised aspartase and transaminase for high-yield synthesis of l-phenylalanine
Abstract

•One-pot synthesis of l-phenylalanine catalysed by coupling the enzymes aspartase and transaminase.•Enzymes were compatible upon reaction and immobilisation conditions.•Concentration of substrates and enzymes were optimised.•Co-immobilisation of enzymes allowed the synthesis of high concentrations of Phe.

l-phenylalanine (Phe) was synthesised by coupling the enzymes aspartase (AspB) catalysing the synthesis of l-aspartate from fumarate and NH4Cl and microbial aspartate transaminase (TA) catalysing the transfer of the amino group from l-aspartate to phenylpyruvate. Phe synthesis was studied with enzymes in solution and immobilised separately and together on amino-epoxy Relizyme® support. Immobilisation efficiencies and recovered activities of co-immobilised enzymes were slightly lower than those obtained when immobilised separately. Substrate and enzyme concentrations for the synthesis reactions were optimised as follows: co-immobilised 0.3 U/mL AspB and 2 U/mL TA, 0.15 M fumarate, 0.3 M NH4Cl, 0.1 M phenylpyruvate, 0.1 mM pyridoxal-5′-phosphate (PLP) at pH 7.5 and 37 °C. Total reaction yield of 83% and Phe yield of 95% were obtained. The initial rates of the reactions catalysed by co-immobilised enzymes were similar to those obtained when the reactions were catalysed by free enzymes, indicating negligible diffusional limitations associated to the application of the co-immobilised enzymes.

Keywords
One-pot multienzymatic reaction; Co-immobilised enzymes; l-phenylalanine; Aspartase; Transaminase; Immobilisation on epoxy support.AspB, Laspartate ammonia-lyase (EC 4.3.1.1) from Bacillus sp. YM55-1; AAT, Laspartate transaminase (EC 2.6.1.1) from porcine
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Co-immobilised aspartase and transaminase for high-yield synthesis of l-phenylalanine
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 93, 15 January 2015, Pages 173–178
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