Efficient gamma-aminobutyric acid bioconversion by engineered <i>Escherichia coli</i>

Ke, Chongrong; Wei, Jie; Ren, Yang; Yang, Xinwei; Chen, Jia; Huang, Jianzhong

doi:10.1080/13102818.2018.1446765

Cited by 8 publications

(4 citation statements)

References 29 publications

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“…Several mutational approaches such as directed evolution and site-specific mutagenesis are considered as powerful tools for optimizing or improving enzyme properties. Several researchers have applied these approaches to improve GAD activity [84,[97][98][99][100][101] and were applied in whole-cell biocatalysts. In order to improve GAD activity over an expanded pH range, recombinant C. glutamicum cells were obtained by expressing L. brevis Lb85 GadB variants.…”

Section: Improvement Of Gad Activities and Gaba Productionmentioning

confidence: 99%

Glutamate Decarboxylase from Lactic Acid Bacteria—A Key Enzyme in GABA Synthesis

2020

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Glutamate decarboxylase (l-glutamate-1-carboxylase, GAD; EC 4.1.1.15) is a pyridoxal-5’-phosphate-dependent enzyme that catalyzes the irreversible α-decarboxylation of l-glutamic acid to γ-aminobutyric acid (GABA) and CO2. The enzyme is widely distributed in eukaryotes as well as prokaryotes, where it—together with its reaction product GABA—fulfils very different physiological functions. The occurrence of gad genes encoding GAD has been shown for many microorganisms, and GABA-producing lactic acid bacteria (LAB) have been a focus of research during recent years. A wide range of traditional foods produced by fermentation based on LAB offer the potential of providing new functional food products enriched with GABA that may offer certain health-benefits. Different GAD enzymes and genes from several strains of LAB have been isolated and characterized recently. GABA-producing LAB, the biochemical properties of their GAD enzymes, and possible applications are reviewed here.

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Section: Improvement Of Gad Activities and Gaba Productionmentioning

confidence: 99%

Glutamate Decarboxylase from Lactic Acid Bacteria—A Key Enzyme in GABA Synthesis

2020

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“…The optimal induction conditions for the whole-cell biocatalyst were explored using a single-factor experimental design. Recombinant cells were subjected to induction at various temperatures (16, 20, 25, 30, and 37°C) for varying durations (2, 4, 6, 10, and 16 h) using 1 mmol/L IPTG [40]. The experiments were replicated three times for each set.…”

Section: Construction Of Plasmids and Strainsmentioning

confidence: 99%

Efficient Whole-cell Catalysis for γ-Aminobutyric Acid Production Using Engineered Escherichia coli

Chang,

Wang,

Zhang

et al. 2023

Preprint

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Background γ-Aminobutyric acid (GABA) is a non-proteinogenic amino acid that has extensive applications in the food, feed, pharmaceutical, and chemical synthesis fields. The utilization of engineered Escherichia coli in whole-cell catalysis offers a promising approach for GABA synthesis based on the rapid reaction kinetics and reduced byproduct formation. Previously, we constructed a recombinant E. coli that efficiently converts L-glutamate (L-Glu) to GABA; however, freezing and thawing of the strain and the addition of external pyridoxal 5′-phosphate (PLP) were required. The objective of this investigation was to enhance the efficiency of GABA synthesis through E. coli genetic modifications to achieve a more streamlined production process. Results First, the inducible expression conditions of the gad gene were optimized to 37°C for 6 h. Next, cell permeability was improved by overexpressing sulA in E. coli, which eliminated the need for the freeze-thaw treatment during GABA production. The overexpression of pdxS and pdxT from Bacillus subtilis strain 168 resulted in an ideal engineered strain without the addition of external PLP. Thus, an efficient whole-cell biocatalytic process was optimized. The ideal isopropyl β-D-thiogalactopyranoside concentration, cellular density, and reaction temperature were 0.2 mmol/L, 15 units, and 37°C, respectively, and the substrate consisted of a 4:1 ratio of L-glutamic acid (L-Glu) to L-monosodium glutamate (L-MSG). Ultimately, the optimized conditions were employed for a bioconversion procedure using whole cells in a 3 L bioreactor. The microbial strain was capable of being utilized for a minimum of two cycles with 1 mol/L substrate mixtures, thus achieving a GABA productivity of 103.1 g/L/h and a molar yield of 100.0%. Conclusion A whole-cell catalyst for highly efficient GABA production from a mixture of L-Glu and L-MSG was constructed by engineering E. coli, and the freeze-thaw steps and external PLP addition were not required. This research illustrates that the recently engineered strain of E. coli exhibits promise for utilization in the large-scale industrial synthesis of GABA.

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“…Several mutational approaches such as directed evolution and site-specific mutagenesis are considered as powerful tools for optimizing or improving enzyme properties. Several researchers have applied these approaches to improve GAD activity [83,[97][98][99][100] that was applied in whole-cell biocatalysts. In order to improve GAD activity over an expanded pH range, recombinant C. glutamicum cells were obtained by expressing L. brevis Lb85 GadB variants.…”

Section: Biochemical Insights Into Glutamate Decarboxylase From Lactimentioning

confidence: 99%

A Brief Review on Glutamate Decarboxylase From Lactic Acid Bacteria

Yogeswara¹,

Maneerat²,

Haltrich³

2020

Preprint

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Glutamate decarboxylase (L-glutamate-1-carboxylase, GAD; EC 4.1.1.15) is a pyridoxal 5-phosphate-dependent enzyme, which catalyzes the irreversible α-decarboxylation of L-glutamic acid to γ-aminobutyric acid (GABA) and CO2. The enzyme is widely distributed in eukaryotes as well as prokaryotes, where it – together with its reaction product GABA - fulfils very different physiological functions. The occurrence of gad genes encoding GAD has been shown for many microorganisms, and GABA-producing lactic acid bacteria (LAB) have been a focus of research during recent years. A wide range of traditional foods produced by fermentation based on LAB offer the potential of providing new functional food products enriched with GABA that may offer certain health-benefits. Different GAD enzymes and genes from several strains of LAB have been isolated and characterized recently. GABA-producing LAB, biochemical properties of their GAD enzymes, and possible applications are reviewed here.

show abstract

Efficient gamma-aminobutyric acid bioconversion by engineered Escherichia coli

Cited by 8 publications

References 29 publications

Glutamate Decarboxylase from Lactic Acid Bacteria—A Key Enzyme in GABA Synthesis

Glutamate Decarboxylase from Lactic Acid Bacteria—A Key Enzyme in GABA Synthesis

Efficient Whole-cell Catalysis for γ-Aminobutyric Acid Production Using Engineered Escherichia coli

A Brief Review on Glutamate Decarboxylase From Lactic Acid Bacteria

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