Knockout of the Aldehyde Dehydrogenase Gene in Fusarium oxysporum for Enhanced Ethanol Yield

Jinxia Fan, Xiaomei Huang, Guoxiang Zheng, Changyu Liu, Ming Wang, Yong Sun and Qian Yang ()
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Jinxia Fan: College of Engineering, Northeast Agricultural University, Harbin 150030, China
Xiaomei Huang: Department of Bioengineering, Heilongjiang Vocational College of Agricultural Technology, Jiamusi 154007, China
Guoxiang Zheng: College of Engineering, Northeast Agricultural University, Harbin 150030, China
Changyu Liu: College of Engineering, Northeast Agricultural University, Harbin 150030, China
Ming Wang: College of Engineering, Northeast Agricultural University, Harbin 150030, China
Yong Sun: College of Engineering, Northeast Agricultural University, Harbin 150030, China
Qian Yang: Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China

Energies, 2022, vol. 16, issue 1, 1-12

Abstract: Acetic acid is the primary by-product generated from ethanol production by Fusarium oxysporum using glucose or xylose as a substrate. Aldehyde dehydrogenase (ALDH) is the critical enzyme in acetic acid metabolism. To decrease acetic acid yield in ethanol production, the 1509 bp DNA of aldh , encoding a 502 amino acid protein with a calculated molecular mass of 54.33 kDa and an isoelectric point of 6.21, was cloned from F. oxysporum . Sequence analysis confirmed that the screened proteins belonged to the ALDH family. A knockout vector, ∆ aldh , containing positive (hygromycin resistance gene) and negative (thymidine kinase gene from the herpes simplex virus) selectable markers, was constructed. Ethanol production by the mutant (cs28pCAM-Pstal-∆ aldh ) in glucose- and xylose-containing media was 0.46 and 0.39 g/g, respectively, and these yields were 16.93% and 34.63% higher than those by the wild-type strain (0.393 and 0.289 g/g). Furthermore, the acetic acid yield of the mutant was 3.50 and 3.01 g/L, respectively, showing a 23.10% and 39.55% decrease compared with the wild-type strain (4.308 and 4.196 g/L). The biomass of the mutant (4.05 and 4.52 g/L) was lower than that of the wild-type strain (4.71 and 5.97 g/L). These results demonstrated the potential use of the genetically stable mutant for industrial bioethanol production.

Keywords: aldehyde dehydrogenase; Fusarium oxysporum; gene knockout; acetic acid (search for similar items in EconPapers)
JEL-codes: Q Q0 Q4 Q40 Q41 Q42 Q43 Q47 Q48 Q49 (search for similar items in EconPapers)
Date: 2022
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