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从山羊瘤胃微生物组中鉴定和功能表达新型木酮糖异构酶在酿酒酵母中。

Identification and functional expression of a new xylose isomerase from the goat rumen microbiome in Saccharomyces cerevisiae.

机构信息

Genetics and Biotechnology Laboratory, Embrapa-Agroenergy, Brasília, DF, Brazil.

Genomic Sciences and Biotechnology Program, Universidade Católica de Brasília, Brasília, DF, Brazil.

出版信息

Lett Appl Microbiol. 2022 Jun;74(6):941-948. doi: 10.1111/lam.13689. Epub 2022 Mar 8.

Abstract

The current climate crisis demands replacement of fossil energy sources with sustainable alternatives. In this scenario, second-generation bioethanol, a product of lignocellulosic biomass fermentation, represents a more sustainable alternative. However, Saccharomyces cerevisiae cannot metabolize pentoses, such as xylose, present as a major component of lignocellulosic biomass. Xylose isomerase (XI) is an enzyme that allows xylose consumption by yeasts, because it converts xylose into xylulose, which is further converted to ethanol by the pentose-phosphate pathway. Only a few XI were successfully expressed in S. cerevisiae strains. This work presents a new bacterial XI, named GR-XI 1, obtained from a Brazilian goat rumen metagenomic library. Phylogenetic analysis confirmed the bacterial origin of the gene, which is related to Firmicutes XIs. After codon optimization, this enzyme, renamed XySC1, was functionally expressed in S. cerevisiae, allowing growth in media with xylose as sole carbon source. Overexpression of XySC1 in S. cerevisiae allowed the recombinant strain to efficiently consume and metabolize xylose under aerobic conditions.

摘要

当前的气候危机要求用可持续的替代品替代化石能源。在此背景下,第二代生物乙醇,一种木质纤维素生物质发酵的产物,代表了一种更可持续的选择。然而,酿酒酵母不能代谢戊糖,如木质纤维素生物质的主要成分木糖。木糖异构酶(XI)是一种允许酵母消耗木糖的酶,因为它将木糖转化为木酮糖,木酮糖进一步通过磷酸戊糖途径转化为乙醇。只有少数几个 XI 在酿酒酵母菌株中成功表达。这项工作介绍了一种新的细菌 XI,名为 GR-XI 1,它是从巴西山羊瘤胃宏基因组文库中获得的。系统发育分析证实了该基因的细菌起源,它与厚壁菌门的 XI 有关。经过密码子优化后,这种酶被重新命名为 XySC1,在酿酒酵母中得到了功能性表达,使其能够在以木糖为唯一碳源的培养基中生长。在酿酒酵母中过表达 XySC1 使重组菌株能够在有氧条件下高效消耗和代谢木糖。

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