Expression and characterization of monofunctional alcohol dehydrogenase enzymes in Clostridium thermocellum

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2024-06-20 DOI:10.1016/j.mec.2024.e00243
Daniela Prates Chiarelli , Bishal Dev Sharma , Shuen Hon , Luana Walravens Bergamo , Lee R. Lynd , Daniel G. Olson
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Abstract

Clostridium thermocellum is a thermophilic anaerobic bacterium that could be used for cellulosic biofuel production due to its strong native ability to consume cellulose, however its ethanol production ability needs to be improved to enable commercial application. In our previous strain engineering work, we observed a spontaneous mutation in the native adhE gene that reduced ethanol production. Here we attempted to complement this mutation by heterologous expression of 18 different alcohol dehydrogenase (adh) genes. We were able to express all of them successfully in C. thermocellum. Surprisingly, however, none of them increased ethanol production, and several actually decreased it. Our findings contribute to understanding the correlation between C. thermocellum ethanol production and Adh enzyme cofactor preferences. The identification of a set of adh genes that can be successfully expressed in this organism provides a foundation for future investigations into how the properties of Adh enzymes affect ethanol production.

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热梭菌中单功能醇脱氢酶的表达和特性分析
嗜热梭菌是一种嗜热厌氧细菌,由于其具有很强的消耗纤维素的原生能力,可用于纤维素生物燃料的生产。在我们之前的菌株工程工作中,我们观察到原生 adhE 基因发生了自发突变,从而降低了乙醇产量。在此,我们尝试通过异源表达 18 种不同的乙醇脱氢酶(adh)基因来补充这一突变。我们成功地在热菌中表达了所有这些基因。但令人惊讶的是,这些基因都没有提高乙醇产量,其中几个基因实际上还降低了乙醇产量。我们的发现有助于理解热菌乙醇产量与 Adh 酶辅因子偏好之间的相关性。确定了一组可在该生物体内成功表达的 Adh 基因,为今后研究 Adh 酶的特性如何影响乙醇产量奠定了基础。
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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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