Efficient expression of novel glutamate decarboxylases and high level production of γ-aminobutyric acid catalyzed by engineered Escherichia coli

Tang, Cun-Duo; Li, Xiang; Shi, Hongbo; Jia, Yuanyuan; Dong, Zixing; Jiao, Zhujin; Wang, Linfeng; Xu, Jian‐He; Yao, Lunguang; Kan, Yunchao

doi:10.1016/j.ijbiomac.2020.05.195

Cited by 27 publications

(23 citation statements)

References 35 publications

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“… 21 , 28 , 29 , 35 Recently, there has been an attempt to search for novel GADs from GRAS probiotic bacteria including LAB based on genome mining and sequence alignment analysis, and consequently, 16 putative GADs were screened. 29 Among them, LsGAD and LlGAD were found to possess a wider optimal pH range and a more durable enzymatic stability than GAD from L. brevis , which was used as a probe. 29 Therefore, in this study, we hereby examined three different GADs (EcGADmut, LsGAD, and LlGAD) that were previously reported to exhibit superior activity under a wide pH range (pH 5.0–7.0) for GABA production from glucose at near-neutral pH in l -glutamate-overproducing C. glutamicum KCTC 1852 strains.…”

Section: Resultsmentioning

confidence: 99%

“…First, the GABA biosynthesis pathway was established in recombinant C. glutamicum KCTC 1852 strains via the construction of plasmids for heterologous expression of three GADs under the control of two different strong synthetic promoters, H30 and H36 ( Table 2 ). 29 , 30 Then, further control of pH in flask cultivation by the intermittent addition of an acid or a base was implemented to investigate the optimal pH for GABA production. Finally, batch fermentation of the engineered strains with heterologous expression of EcGADmut, LsGAD, and LlGAD under the H36 promoter was conducted by maintaining the culture pH at 6.0 and 7.0 ( Figure 1 ).…”

Section: Introductionmentioning

confidence: 99%

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Production of γ-Aminobutyrate (GABA) in Recombinant Corynebacterium glutamicum by Expression of Glutamate Decarboxylase Active at Neutral pH

et al. 2022

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γ-Aminobutyrate (GABA) is an important chemical by itself and can be further used for the production of monomer used for the synthesis of biodegradable polyamides. Until now, GABA production using Corynebacterium glutamicum harboring glutamate decarboxylases (GADs) has been limited due to the discrepancy between optimal pH for GAD activity (pH 4.0) and cell growth (pH 7.0). In this study, we developed recombinant C. glutamicum strains expressing mutated GAD from Escherichia coli (EcGADmut) and GADs from Lactococcus lactis CICC20209 (LlGAD) and Lactobacillus senmaizukei (LsGAD), all of which showed enhanced pH stability and adaptability at a pH of approximately 7.0. In shake flask cultivations, the GABA productions of C. glutamicum H36EcGADmut, C. glutamicum H36LsGAD, and C. glutamicum H36LlGAD were examined at pH 5.0, 6.0, and 7.0, respectively. Finally, C. glutamicum H36EcGADmut (40.3 and 39.3 g L –1 ), H36LlGAD (42.5 and 41.1 g L –1 ), and H36LsGAD (41.6 and 40.2 g L –1 ) produced improved GABA titers and yields in batch fermentation at pH 6.0 and pH 7.0, respectively, from 100 g L –1 glucose. The recombinant strains developed in this study could be used for the establishment of sustainable direct fermentative GABA production from renewable resources under mild culture conditions, thus increasing the availability of various GADs.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Production of γ-Aminobutyrate (GABA) in Recombinant Corynebacterium glutamicum by Expression of Glutamate Decarboxylase Active at Neutral pH

et al. 2022

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“…Besides GABACls channels, other components from the GABAergic system can represent molecular targets, but that is still too little scientifically explored (Ilg et al ., 2013). This system is composed of four primary parts: GABA synthesis by GAD action, GABA catabolism by GABA-transaminase, GABA transporters and GABA receptors (Dionisio et al ., 2011; Ilg et al ., 2013; Tang et al ., 2020).…”

Section: Resultsmentioning

confidence: 99%

“…GAD is a key enzyme in the dynamic regulation of neural network excitability because both its substrate and product are neurotransmitters and exhibit opposite actions ( l -glutamate – excitatory effect and GABA – inhibitory action). GAD is considered as an initiator on the GABAergic system and is frequently used as a molecular marker from GABAergic neurons (Li et al ., 2016; Tang et al ., 2020).…”

Section: Resultsmentioning

confidence: 99%

Molecular targets for the development of new acaricides againstRhipicephalus microplus: a review

et al. 2022

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The cattle tick Rhipicephalus microplus is an ectoparasite with high economic importance to bovine culture, mainly in tropical and subtropical regions. The resistance of the tick from the commercial acaricides has hindered its control, thus motivating the search for new strategies. The purpose of this study was to perform a critical review about the main molecular targets of R. microplus that are useful for the discovery of new acaricides. Bibliographic search was conducted in the databases PubMed, ScienceDirect and CAB Direct, using the following descriptors: ‘Rhipicephalus microplus’, ‘Boophilus microplus’, ‘molecular targets’ and ‘action’, published between 2010 and 2021. Out of the 212 publications identified, 17 articles were selected for study inclusion. This review described 14 molecular targets and among these 4 are targets from commercial acaricides. Most of them are enzymes to catalyse important reactions to tick survival, related to energetic metabolism, mechanisms of biotransformation and neurotransmission. The data will be helpful in the development of new more effective and selective acaricides.

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“…Glutamate decarboxylase (GAD) has the potential of converting l ‐glutamate to form GABA (Tang et al, 2020). Corynebacterium glutamicum G01 efficiently converts glucose to the l ‐glutamate, but it cannot yield GABA from l ‐glutamate due to the lack of glutamate decarboxylase and pyridoxal kinase (Ham et al, 2022).…”

Section: Biosynthesis Of Chemicals Which Plp Participated As Cofactormentioning

confidence: 99%

Advances and prospects in the biosynthesis of vitamin B₆ and its bioengineering as a cofactor of other chemicals

Liu

Jin

et al. 2022

Food Bioengineering

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Vitamin B6 is an essential nutrient, which is widely used in food products, feed additives, pharmaceuticals, disease diagnosis, and other fields. Pyridoxal‐5′‐phosphate (PLP), the active form of vitamin B6, is an important coenzyme participating in a variety of enzyme reactions. At present, the oxazole method is mainly used for the production of vitamin B6, but toxic and corrosive intermediates produced in the synthesis process, which does not conform to the green manufacturing. Therefore, there is considerable interest in shifting from chemical processes to sustainable fermentation techniques and research on PLP metabolism of other valuable compounds. In this review, we will briefly describe the enzymes that PLP participated and focus on the advances in vitamin B6 biosynthesis and discuss its application to engineering bacteria that overproduce other commercially valuable substances including cadaverine, 3,4‐dihydroxyphenylacetate‐l‐alanine, γ‐aminobutyric acid, and l‐phosphinothricin. It will provide a reference for the biosynthesis of vitamin B6 or other valuable compounds in which PLP participates in the reaction, and we also look forward to the future development prospect of PLP.

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Efficient expression of novel glutamate decarboxylases and high level production of γ-aminobutyric acid catalyzed by engineered Escherichia coli

Cited by 27 publications

References 35 publications

Production of γ-Aminobutyrate (GABA) in Recombinant Corynebacterium glutamicum by Expression of Glutamate Decarboxylase Active at Neutral pH

Production of γ-Aminobutyrate (GABA) in Recombinant Corynebacterium glutamicum by Expression of Glutamate Decarboxylase Active at Neutral pH

Molecular targets for the development of new acaricides againstRhipicephalus microplus: a review

Advances and prospects in the biosynthesis of vitamin B₆ and its bioengineering as a cofactor of other chemicals

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Efficient expression of novel glutamate decarboxylases and high level production of γ-aminobutyric acid catalyzed by engineered Escherichia coli

Cited by 27 publications

References 35 publications

Production of γ-Aminobutyrate (GABA) in Recombinant Corynebacterium glutamicum by Expression of Glutamate Decarboxylase Active at Neutral pH

Production of γ-Aminobutyrate (GABA) in Recombinant Corynebacterium glutamicum by Expression of Glutamate Decarboxylase Active at Neutral pH

Molecular targets for the development of new acaricides againstRhipicephalus microplus: a review

Advances and prospects in the biosynthesis of vitamin B6 and its bioengineering as a cofactor of other chemicals

Contact Info

Product

Resources

About

Advances and prospects in the biosynthesis of vitamin B₆ and its bioengineering as a cofactor of other chemicals