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Enhancement of Gluconobacter oxydans Resistance to Lignocellulosic-Derived Inhibitors in Xylonic Acid Production by Overexpressing Thioredoxin

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Abstract

Efficient utilization of lignocellulose is an economically relevant practice for improving the financial prospects of biorefineries. Lignocellulose contains significant levels of xylose that can be converted into valuable xylonic acid. However, some inhibitors of bioconversion processes are produced after pretreatment. Xylonic acid production in bacteria, such as Gluconobacter oxydans, is hindered by poor bacterial tolerance to contaminants. Therefore, in order to enhance bacterial resistance to inhibitors, a recombinant strain of G. oxydans was created by the introduction of the thioredoxin gene. Thioredoxin is a key protein responsible for maintaining cellular redox potential and is critical to the conversion of xylose to xylonate. Overexpression of thioredoxin was confirmed at the enzymatic level, while the recombinant strain showed increased catalytic activity when inhibitors, such as formic acid or p-hydroxybenzaldehyde (PHBA), were added to the synthetic xylose medium (17% and 7% improvement in xylonic acid yield, respectively). To probe the molecular mechanism behind the recombinant strain response to inhibitors, the expression levels of various genes were analyzed by qRT-PCR, which revealed five differentially expressed genes (DEGs) upon exposure to formic acid or PHBA.

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Funding

The research was supported by the National Natural Science Foundation of China (31370573, 31901270). Also, the authors gratefully acknowledge the financial support from the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)

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Authors and Affiliations

  1. Key Laboratory of Forestry Genetics & Biotechnology, Nanjing Forestry University, Ministry of Education, Nanjing, 210037, People’s Republic of China

    Yi Shen, Xin Zhou & Yong Xu

  2. Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, No. 159 Longpan Road, Nanjing, 210037, People’s Republic of China

    Yi Shen, Xin Zhou & Yong Xu

  3. Nanjing Santa Scott Biotechnology Co., Ltd., Nanjing, 211100, People’s Republic of China

    Yi Shen

  4. Jiangsu Province Key Laboratory of Green Biomass-based Fuels and Chemicals, Nanjing, 210037, People’s Republic of China

    Xin Zhou & Yong Xu

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  1. Yi Shen
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Correspondence to Yong Xu.

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Electronic Supplementary Material

ESM 1

Relative quantity of 19 DEGs in recombinant strains with 3 inhibitors at 24 h (JPG 1983 kb)

ESM 2

Relative quantity of 19 DEGs in recombinant strains with 3 inhibitors at 48 h (JPG 1950 kb)

ESM 3

Relative quantity of 19 DEGs in recombinant strains with 3 inhibitors at 72 h (JPG 1857 kb)

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Shen, Y., Zhou, X. & Xu, Y. Enhancement of Gluconobacter oxydans Resistance to Lignocellulosic-Derived Inhibitors in Xylonic Acid Production by Overexpressing Thioredoxin. Appl Biochem Biotechnol 191, 1072–1083 (2020). https://doi.org/10.1007/s12010-020-03253-6

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  • DOI: https://doi.org/10.1007/s12010-020-03253-6

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