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对酿酒酵母进行工程改造以实现L-阿拉伯糖的高效厌氧酒精发酵。

Engineering of Saccharomyces cerevisiae for efficient anaerobic alcoholic fermentation of L-arabinose.

作者信息

Wisselink H Wouter, Toirkens Maurice J, del Rosario Franco Berriel M, Winkler Aaron A, van Dijken Johannes P, Pronk Jack T, van Maris Antonius J A

机构信息

Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands.

出版信息

Appl Environ Microbiol. 2007 Aug;73(15):4881-91. doi: 10.1128/AEM.00177-07. Epub 2007 Jun 1.

Abstract

For cost-effective and efficient ethanol production from lignocellulosic fractions of plant biomass, the conversion of not only major constituents, such as glucose and xylose, but also less predominant sugars, such as l-arabinose, is required. Wild-type strains of Saccharomyces cerevisiae, the organism used in industrial ethanol production, cannot ferment xylose and arabinose. Although metabolic and evolutionary engineering has enabled the efficient alcoholic fermentation of xylose under anaerobic conditions, the conversion of l-arabinose into ethanol by engineered S. cerevisiae strains has previously been demonstrated only under oxygen-limited conditions. This study reports the first case of fast and efficient anaerobic alcoholic fermentation of l-arabinose by an engineered S. cerevisiae strain. This fermentation was achieved by combining the expression of the structural genes for the l-arabinose utilization pathway of Lactobacillus plantarum, the overexpression of the S. cerevisiae genes encoding the enzymes of the nonoxidative pentose phosphate pathway, and extensive evolutionary engineering. The resulting S. cerevisiae strain exhibited high rates of arabinose consumption (0.70 g h(-1) g dry weight) and ethanol production (0.29 g h(-1) g dry weight) and a high ethanol yield (0.43 g g(-1)) during anaerobic growth on l-arabinose as the sole carbon source. In addition, efficient ethanol production from sugar mixtures containing glucose and arabinose, which is crucial for application in industrial ethanol production, was achieved.

摘要

为了从植物生物质的木质纤维素部分经济高效地生产乙醇,不仅需要转化主要成分,如葡萄糖和木糖,还需要转化不太主要的糖类,如L-阿拉伯糖。工业乙醇生产中使用的酿酒酵母野生型菌株不能发酵木糖和阿拉伯糖。尽管代谢工程和进化工程已使木糖在厌氧条件下能够进行高效酒精发酵,但此前工程酿酒酵母菌株将L-阿拉伯糖转化为乙醇仅在限氧条件下得到证实。本研究报道了首例工程酿酒酵母菌株对L-阿拉伯糖进行快速高效厌氧酒精发酵的案例。这种发酵是通过将植物乳杆菌L-阿拉伯糖利用途径的结构基因表达、编码非氧化戊糖磷酸途径酶的酿酒酵母基因过表达以及广泛的进化工程相结合来实现的。所得的酿酒酵母菌株在以L-阿拉伯糖作为唯一碳源的厌氧生长过程中,表现出较高的阿拉伯糖消耗速率(0.70 g h(-1) g 干重)和乙醇生产速率(0.29 g h(-1) g 干重)以及较高的乙醇产率(0.43 g g(-1))。此外,还实现了从含有葡萄糖和阿拉伯糖的糖混合物中高效生产乙醇,这对于工业乙醇生产的应用至关重要。

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