Engineering of a new Escherichia coli strain efficiently metabolizing cellobiose with promising perspectives for plant biomass-based application design

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2021-06-01 DOI:10.1016/j.mec.2020.e00157
Romain Borne , Nicolas Vita , Nathalie Franche , Chantal Tardif, Stéphanie Perret, Henri-Pierre Fierobe
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引用次数: 2

Abstract

The necessity to decrease our fossil energy dependence requests bioprocesses based on biomass degradation. Cellobiose is the main product released by cellulases when acting on the major plant cell wall polysaccharide constituent, the cellulose. Escherichia coli, one of the most common model organisms for the academy and the industry, is unable to metabolize this disaccharide. In this context, the remodeling of E. coli to catabolize cellobiose should thus constitute an important progress for the design of such applications. Here, we developed a robust E. coli strain able to metabolize cellobiose by integration of a small set of modifications in its genome. Contrary to previous studies that use adaptative evolution to achieve some growth on this sugar by reactivating E. coli cryptic operons coding for cellobiose metabolism, we identified easily insertable modifications impacting the cellobiose import (expression of a gene coding a truncated variant of the maltoporin LamB, modification of the expression of lacY encoding the lactose permease) and its intracellular degradation (genomic insertion of a gene encoding either a cytosolic β-glucosidase or a cellobiose phosphorylase). Taken together, our results provide an easily transferable set of mutations that confers to E. coli an efficient growth phenotype on cellobiose (doubling time of 2.2 ​h in aerobiosis) without any prior adaptation.

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一株高效代谢纤维素二糖的新型大肠杆菌的工程设计及其在植物生物量应用设计中的前景
减少我们对化石能源依赖的必要性要求基于生物质降解的生物过程。纤维素二糖是纤维素酶作用于植物细胞壁的主要多糖成分纤维素时所释放的主要产物。大肠杆菌是学术界和工业界最常见的模式生物之一,它无法代谢这种双糖。在这种情况下,重塑大肠杆菌以分解纤维素糖应该是设计此类应用的重要进展。在这里,我们开发了一种强大的大肠杆菌菌株,能够通过整合其基因组中的一小组修饰来代谢纤维二糖。与先前的研究相反,通过重新激活编码纤维素二糖代谢的大肠杆菌隐操作子,利用适应性进化来实现这种糖的一些生长,我们发现了容易插入的修饰,影响纤维素二糖的进口(编码麦芽糖蛋白LamB的截断变体的基因的表达)。编码乳糖渗透酶的lacY表达的修饰及其细胞内降解(编码胞质β-葡萄糖苷酶或纤维素二糖磷酸化酶的基因的基因组插入)。综上所述,我们的研究结果提供了一组易于转移的突变,这些突变赋予了大肠杆菌在纤维素二糖上的高效生长表型(在有氧状态下翻倍时间为2.2 h),而无需任何预先适应。
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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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