Efficient production of gamma‐aminobutyric acid by engineered Saccharomyces cerevisiae with glutamate decarboxylases from Streptomyces

Biotechnology and Applied Biochemistry
2020.0

Abstract

<jats:title>Abstract</jats:title><jats:p>Gamma‐aminobutyric acid (GABA) is an industrially valuable natural product. This study was aimed to establish an efficient food‐grade production process of GABA by engineering <jats:italic>Saccharomyces cerevisiae</jats:italic> that is generally recognized as safe (GRAS). GABA can be produced by catalytic decarboxylation of <jats:sc>l</jats:sc>‐glutamate (<jats:sc>l</jats:sc>‐Glu) by glutamate decarboxylase (GAD, EC4.1.1.15). Two GADs, SsGAD from <jats:italic>Streptomyces sp</jats:italic>. MJ654‐NF4 and ScGAD from <jats:italic>Streptomyces chromofuscus</jats:italic> ATCC 49982, were heterologously expressed in <jats:italic>S. cerevisiae</jats:italic> BJ5464. The engineered yeast strains were used as whole‐cell biocatalysts for GABA production. <jats:italic>S. cerevisiae</jats:italic> BJ5464/SsGAD exhibited significantly higher efficient catalytic activity than that of <jats:italic>S. cerevisiae</jats:italic> BJ5464/ScGAD. The optimal bioconversion system consisted of a cell density of OD<jats:sub>600</jats:sub> 30, 0.1 M <jats:sc>l</jats:sc>‐Glu, and 0.28 mM pyridoxal phosphate in 0.2 M Na<jats:sub>2</jats:sub>HPO<jats:sub>4</jats:sub>–citric acid buffer with pH 5.4, and the reactions were performed at 50 °C for 12 H. <jats:italic>S. cerevisiae</jats:italic> BJ5464/SsGAD cells can be reused, and the accumulated GABA titer reached 62.6 g/L after 10 batches with an overall molar conversion rate of 60.8 mol%. This work thus provides an effective production process of GABA using engineered yeast for food and pharmaceutical applications.

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