Microbial chassis design and engineering for production of amino acids used in food industry

Wang, Jianli; Ma, Wenjian; Zhou, Jingwen; Wang, Xiaoyuan

doi:10.1007/s43393-022-00137-0

Cited by 5 publications

(3 citation statements)

References 120 publications

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“…In particular, it is a major building block of the biodegradable polyamide nylon 4 and to have antihypertensive and antidepressant effects as a food ingredient. 28,29 Whole-cell catalysis is the primary method for producing GABA, 30,31 which relies on the expression of glutamate decarboxylase (GAD) to convert glutamate to GABA (Figure 4a). We cloned the gadB gene encoding GAD from E. coli into the low-and medium-copy-number T7-BOOST plasmids to assess their protein expression capability.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…The nonprotein amino acid GABA has numerous applications in feed, pharmaceuticals, food, and industrial biotechnology. In particular, it is a major building block of the biodegradable polyamide nylon 4 and to have antihypertensive and antidepressant effects as a food ingredient. , Whole-cell catalysis is the primary method for producing GABA, , which relies on the expression of glutamate decarboxylase (GAD) to convert glutamate to GABA (Figure a). We cloned the gadB gene encoding GAD from E.…”

Section: Resultsmentioning

confidence: 99%

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Efficient Protein Expression and Biosynthetic Gene Cluster Regulation in Bacillus subtilis Driven by a T7-BOOST System

Wu,

Li,

Zhang

et al. 2023

ACS Synth. Biol.

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Bacillus subtilis is a generally recognized as safe microorganism that is widely used for protein expression and chemical production, but has a limited number of genetic regulatory components compared with the Gram-negative model microorganism Escherichia coli. In this study, a two-module plug-and-play T7-based optimized output strategy for transcription (T7-BOOST) systems with low leakage expression and a wide dynamic range was constructed based on the inducible promoters P hy‑spank and P xylA . The first T7 RNA polymerase-driven module was seamlessly integrated into the genome based on the CRISPR/Cpf1 system, while the second expression control module was introduced into low, medium, and high copy plasmids for characterization. As a proof of concept, the T7-BOOST systems were successfully employed for whole-cell catalysis production of γ-aminobutyric acid (109.8 g/L with a 98.0% conversion rate), expression of human αS1 casein and human lactoferrin, and regulation of exogenous lycopene biosynthetic gene cluster and endogenous riboflavin biosynthetic gene cluster. Overall, the T7-BOOST system serves as a stringent, controllable, and effective tool for regulating gene expression in B. subtilis.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Efficient Protein Expression and Biosynthetic Gene Cluster Regulation in Bacillus subtilis Driven by a T7-BOOST System

Wu,

Li,

Zhang

et al. 2023

ACS Synth. Biol.

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“…Nonetheless, the industrial-scale production of 2,5-DMP remains a significant challenge, and the yield of 2,5-DMP remains to be improved from the following aspects. First, 2,5-DMP, as a derivative of the l -aspartate family amino acids, can potentially benefit from metabolic engineering strategies commonly used for these amino acids. ,− Moreover, the effect of pH on the fermentation of 2,5-DMP also deserves future research attention . In addition, there are no reports on transport proteins of 2,5-DMP, so the mining of transport proteins is also an urgent issue.…”

Section: Discussionmentioning

confidence: 99%

Metabolic Engineering of Escherichia coli for Production of 2,5-Dimethylpyrazine

Zeng,

Wu,

Han

et al. 2024

J. Agric. Food Chem.

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2,5-Dimethylpyrazine (2,5-DMP) is a high-value-added alkylpyrazine compound with important applications in both the food and pharmaceutical fields. In response to the increasing consumer preference for natural products over chemically synthesized ones, efforts have been made to develop efficient microbial cell factories for the production of 2,5-DMP. However, the previously reported recombinant strains have exhibited low yields and relied on expensive antibiotics and inducers. In this study, we employed metabolic engineering strategies to develop an Escherichia coli strain capable of producing 2,5-DMP at high levels without the need for inducers or antibiotics. Initially, the biosynthesis pathway of 2,5-DMP was constructed that realized 2,5-DMP production from glucose. Subsequently, efforts focused on enhancing 2,5-DMP production by improving the availability of the cofactor NAD + and precursor L-threonine. Additionally, the supply and conversion of L-threonine were balanced by optimizing the copy number of the key gene tdh on the chromosome and by modifying the L-threonine transport system. The final engineering strain D19 produced 3.1 g/L of 2,5-DMP, which is the highest titer for fermentative production of 2,5-DMP using glucose as the carbon source up to date. The strategies used in this study lay a good foundation for the production of 2,5-DMP on a large scale.

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Emerging Technologies for the Recovery of Microbial Bioactive Compounds

Srivastava,

Dasila

2023

Microbial Bioactive Compounds

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Microbial chassis design and engineering for production of amino acids used in food industry

Cited by 5 publications

References 120 publications

Efficient Protein Expression and Biosynthetic Gene Cluster Regulation in Bacillus subtilis Driven by a T7-BOOST System

Efficient Protein Expression and Biosynthetic Gene Cluster Regulation in Bacillus subtilis Driven by a T7-BOOST System

Metabolic Engineering of Escherichia coli for Production of 2,5-Dimethylpyrazine

Emerging Technologies for the Recovery of Microbial Bioactive Compounds

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