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Microbial Cell Factories · 2019 ·Vol. 18 ·期 1 ·1-10 · IF 4.402

Fermentative production of the unnatural amino acid l-2-aminobutyric acid based on metabolic engineering

Xu Jian-Miao,Li Jian-Qiang,Zhang Bo,Liu Zhi-Qiang,Zheng Yu-Guo

Background l -2-aminobutyric acid ( l -ABA) is an unnatural amino acid that is a key intermediate for the synthesis of several important pharmaceuticals. To make the biosynthesis of l -ABA environmental friendly and more suitable for the industrial-scale production. We expand the nature metabolic network of Escherichia coli using metabolic engineering approach for the production of l -ABA. Results In this study, Escherichia coli THR strain with a modified pathway for threonine-hyperproduction was engineered via deletion of the rhtA gene from the chromosome. To redirect carbon flux from 2-ketobutyrate (2-KB) to l -ABA, the ilvIH gene was deleted to block the l -isoleucine pathway. Furthermore, the ilvA gene from Escherichia coli W3110 and the leuDH gene from Thermoactinomyces intermedius were amplified and co-overexpressed. The promoter was altered to regulate the expression strength of ilvA* and leuDH . The final engineered strain E. coli THR Δ rhtA Δ ilvIH /Gap- ilvA *-Pbs- leuDH was able to produce 9.33 g/L of l -ABA with a yield of 0.19 g/L/h by fed-batch fermentation in a 5 L bioreactor. Conclusions This novel metabolically tailored strain offers a promising approach to fulfill industrial requirements for production of l -ABA.

CAS / MDL: 8063-07-8
研究领域: 生物技术 · 代谢工程 · 微生物生物技术
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Foods · 2024 ·Vol. 13 ·期 17 ·2731 · IF 4.7

Characterization of Key Aroma Compounds of Soy Sauce-like Aroma Produced in Ferment of Soybeans by Bacillus subtilis BJ3-2

Qibo Tan,Yongjun Wu,Cen Li,Jing Jin,Lincheng Zhang,Shuoqiu Tong,Zhaofeng Chen,Li Ran,Lu Huang,Zeyan Zuo

Fermented soybeans are popular among many for their rich soy sauce-like aroma. However, the precise composition of this aroma remains elusive, with key aroma compounds unidentified. In this study, we screened the candidate genesilvAandserAin BJ3-2 based on previous multi-omics data, and we constructed three mutant strains, BJ3-2-ΔserA, BJ3-2-ΔilvA, and BJ3-2-ΔserAΔilvA, using homologous recombination to fermented soybeans with varying intensities of soy sauce-like aroma. Our objective was to analyze samples that exhibited different aroma intensities resulting from the fermented soybeans of BJ3-2 and its mutant strains, thereby exploring the key flavor compounds influencing soy sauce-like aroma as well analyzing the effects ofilvAandserAon soy sauce-like aroma. We employed quantitative descriptive sensory analysis (QDA), gas chromatography–olfactometry–mass spectrometry (GC-O-MS), relative odor activity value analysis (rOAV), principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), and partial least squares regression analysis (PLSR). QDA revealed the predominant soy sauce-like aroma profile of roasted and smoky aromas. GC-MS detected 99 volatile components, predominantly pyrazines and ketones, across the four samples, each showing varying concentrations. Based on rOAV (>1) and GC-O, 12 compounds emerged as primary contributors to soy sauce-like aroma. PCA and OPLS-DA were instrumental in discerning aroma differences among the samples, identifying five compounds with VIP > 1 as key marker compounds influencing soy sauce-like aroma intensity levels. Differential analyses of key aroma compounds indicated that the mutant strains ofilvAandserAaffected soy sauce-like aroma mainly by affecting pyrazines. PLSR analysis indicated that roasted and smoky aromas were the two most important sensory attributes of soy sauce-like aroma, with pyrazines associated with roasted aroma and guaiacol associated with smoky aroma. In addition, substances positively correlated with the intensity of soy sauce-like aroma were verified by additional experiments. This study enhances our understanding of the characteristic flavor compounds in soy sauce-like aroma ferments, provides new perspectives for analyzing the molecular mechanisms of soy sauce-like aroma formation, and provides a theoretical framework for the targeted enhancement of soy sauce-like aroma in various foods.

研究领域: 食品科学 · 微生物学 · 分析化学 · 感官分析
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