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用于……的乙酰丙酸诱导型和可调基因表达系统

Levulinic Acid-Inducible and Tunable Gene Expression System for .

作者信息

Sathesh-Prabu Chandran, Ryu Young Shin, Lee Sung Kuk

机构信息

School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, South Korea.

Department of Energy Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, South Korea.

出版信息

Front Bioeng Biotechnol. 2021 Dec 15;9:797020. doi: 10.3389/fbioe.2021.797020. eCollection 2021.

Abstract

AM1 is an efficient platform strain possessing biotechnological potential in formate- and methanol-based single carbon (C1) bioeconomy. Constitutive expression or costly chemical-inducible expression systems are not always desirable. Here, several glucose-, xylose-, and levulinic acid (LA)-inducible promoter systems were assessed for the induction of green fluorescent protein (GFP) as a reporter protein. Among them, the LA-inducible gene expression system (HpdR/P ) showed a strong expression of GFP (51-fold) compared to the control. The system was induced even at a low concentration of LA (0.1 mM). The fluorescence intensity increased with increasing concentrations of LA up to 20 mM. The system was tunable and tightly controlled with meager basal expression. The maximum GFP yield obtained using the system was 42 mg/g biomass, representing 10% of the total protein content. The efficiency of the proposed system was nearly equivalent (90%-100%) to that of the widely used strong promoters such as P and P . The HpdR/P system worked equally efficiently in five different strains of . LA is a low-cost, renewable, and sustainable platform chemical that can be used to generate a wide range of products. Hence, the reported system in potent strains of is highly beneficial in the C1-biorefinery industry to produce value-added products and bulk chemicals

摘要

AM1是一种高效的平台菌株,在基于甲酸盐和甲醇的单碳(C1)生物经济中具有生物技术潜力。组成型表达或成本高昂的化学诱导表达系统并不总是理想的。在此,评估了几种葡萄糖、木糖和乙酰丙酸(LA)诱导的启动子系统对绿色荧光蛋白(GFP)作为报告蛋白的诱导作用。其中,LA诱导基因表达系统(HpdR/P )与对照相比显示出GFP的强烈表达(51倍)。该系统即使在低浓度的LA(0.1 mM)下也能被诱导。荧光强度随着LA浓度增加至20 mM而增加。该系统是可调节的,且基础表达微弱,受到严格控制。使用该系统获得的最大GFP产量为42 mg/g生物量,占总蛋白含量的10%。所提出系统的效率与广泛使用的强启动子如P 和P 的效率几乎相当(90%-100%)。HpdR/P 系统在五种不同的 菌株中同样高效地发挥作用。LA是一种低成本、可再生且可持续的平台化学品,可用于生产多种产品。因此,在 的高效菌株中报道的该系统在C1生物精炼行业中对于生产增值产品和大宗化学品非常有益。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ccf/8714952/04d2e5d1a30b/fbioe-09-797020-g001.jpg

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