利用 CRISPR/Cas9 系统对乳酸菌的各种菌株进行高效代谢途径改造,以提高抗菌化合物的合成。

IF 4.1 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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引用次数: 0

摘要

众所周知,乳酸菌(LAB)在发酵过程中发挥着各种有益作用,可作为益生菌,并产生大量有价值的化合物,包括具有抗菌活性的化合物,其中细菌素样抑制物质(BLIS)可用作生物防腐剂,以提高食品安全和质量。然而,类抑菌物质的产量往往有限,这给目前的保鲜做法带来了商业竞争力方面的挑战。因此,本研究旨在建立一个优化的双质粒 CRISPR/Cas9 系统,通过破坏各种 LAB 菌株上的乳酸脱氢酶基因(ldh),将碳通量从乳酸转向具有抗菌活性的化合物。乳酸缺失(ldhΔ)菌株会导致新陈代谢的转变,从而使其对所选食源性病原体的抑制活性比野生型(WT)菌株提高 78%。效果最明显的是粪肠球菌-ldhΔ,与没有抗菌活性的 WT 菌株相比,它对所有食源性病原体都有明显的杀菌效果。本研究为具有重要经济价值的 LAB 和其他有益细菌合成有价值的食品和工业化合物并提高其产量提供了一个框架模型。本研究首次报道了可以通过改变ldh来操纵某些酵母菌的新陈代谢,从而获得具有更高抗微生物活性的代谢产物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Efficient metabolic pathway modification in various strains of lactic acid bacteria using CRISPR/Cas9 system for elevated synthesis of antimicrobial compounds

Lactic acid bacteria (LAB) are known to exhibit various beneficial roles in fermentation, serving as probiotics, and producing a plethora of valuable compounds including antimicrobial activity such as bacteriocin-like inhibitory substance (BLIS) that can be used as biopreservative to improve food safety and quality. However, the yield of BLIS is often limited, which poses a challenge to be commercially competitive with the current preservation practice. Therefore, the present work aimed to establish an optimised two-plasmid CRISPR/Cas9 system to redirect the carbon flux away from lactate towards compounds with antimicrobial activity by disrupting lactate dehydrogenase gene (ldh) on various strains of LAB. The lactic acid-deficient (ldhΔ) strains caused a metabolic shift resulting in increased inhibitory activity against selected foodborne pathogens up to 78 % than the wild-type (WT) strain. The most significant effect was depicted by Enterococcus faecalis-ldh∆ which displayed prominent bactericidal effects against all foodborne pathogens as compared to the WT that showed no antimicrobial activity. The present work provided a framework model for economically important LAB and other beneficial bacteria to synthesise and increase the yield of valuable food and industrial compounds. The present work reported for the first time that the metabolism of selected LAB can be manipulated by modifying ldh to attain metabolites with higher antimicrobial activity.

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来源期刊
Journal of biotechnology
Journal of biotechnology 工程技术-生物工程与应用微生物
CiteScore
8.90
自引率
2.40%
发文量
190
审稿时长
45 days
期刊介绍: The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.
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