Adaptive laboratory evolution of Bacillus subtilis to overcome toxicity of lignocellulosic hydrolysate derived from Distiller's dried grains with solubles (DDGS)

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2023-06-01 DOI:10.1016/j.mec.2023.e00223
Jasper L.S.P. Driessen , Josefin Johnsen , Ivan Pogrebnyakov , Elsayed T.T. Mohamed , Solange I. Mussatto , Adam M. Feist , Sheila I. Jensen , Alex T. Nielsen
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Abstract

Microbial tolerance to toxic compounds formed during biomass pretreatment is a significant challenge to produce bio-based products from lignocellulose cost effectively. Rational engineering can be problematic due to insufficient prerequisite knowledge of tolerance mechanisms. Therefore, adaptive laboratory evolution was applied to obtain 20 tolerant lineages of Bacillus subtilis strains able to utilize Distiller's Dried Grains with Solubles-derived (DDGS) hydrolysate. Evolved strains showed both improved growth performance and retained heterologous enzyme production using 100% hydrolysate-based medium, whereas growth of the starting strains was essentially absent. Whole-genome resequencing revealed that evolved isolates acquired mutations in the global regulator codY in 15 of the 19 sequenced isolates. Furthermore, mutations in genes related to oxidative stress (katA, perR) and flagella function appeared in both tolerance and control evolution experiments without toxic compounds. Overall, tolerance adaptive laboratory evolution yielded strains able to utilize DDGS-hydrolysate to produce enzymes and hence proved to be a valuable tool for the valorization of lignocellulose.

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枯草芽孢杆菌的适应性实验室进化以克服蒸馏器干燥谷物中木质纤维素水解物的毒性(DDGS)
微生物对生物质预处理过程中形成的有毒化合物的耐受性是从木质纤维素生产生物基产品的一个重大挑战。由于对公差机制的先决条件知识不足,理性工程可能会出现问题。因此,应用适应性实验室进化获得了20个枯草芽孢杆菌菌株的耐受谱系,这些菌株能够利用具有可溶物衍生(DDGS)水解物的Distiller’s Dried Grains。使用100%基于水解产物的培养基,进化菌株显示出提高的生长性能和保留的异源酶产量,而起始菌株基本上没有生长。全基因组重测序显示,进化的分离株在19个测序的分离株中的15个中获得了全球调节因子codY的突变。此外,在没有有毒化合物的耐受和控制进化实验中,都出现了与氧化应激(katA、perR)和鞭毛功能相关的基因突变。总体而言,耐受性适应性实验室进化产生了能够利用DDGS水解物生产酶的菌株,因此被证明是木质纤维素增值的有价值工具。
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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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