Enhancement of glucaric acid production in Saccharomyces cerevisiae by expressing Vitreoscilla hemoglobin
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ORIGINAL RESEARCH PAPER
Enhancement of glucaric acid production in Saccharomyces cerevisiae by expressing Vitreoscilla hemoglobin Xi Zhang . Chi Xu . YingLi Liu . Jing Wang . YunYing Zhao Yu Deng
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Received: 26 March 2020 / Accepted: 11 July 2020 Ó Springer Nature B.V. 2020
Abstract Objective To enhance the glucaric acid (GA) production in Saccharomyces cerevisiae, the Vitreoscilla hemoglobin was employed to reinforce cellular oxygen supplement. Additionally, the pH-free fermentation strategy was engaged to lower the cost brought by base feeding during the acid-accumulated and longperiod glucaric acid production. Results Recombinant yeast Bga-4 was constructed harboring Vitreoscilla hemoglobin on the basis of Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02966-2) contains supplementary material, which is available to authorized users. X. Zhang C. Xu Y. Liu J. Wang Y. Zhao (&) Y. Deng (&) National Engineering Laboratory for Cereal Fermentation Technology (NELCF), School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, People’s Republic of China e-mail: [email protected] Y. Deng e-mail: [email protected] Y. Liu J. Wang China-Canada Joint Lab of Food Nutrition and Health (Beijing), Beijing Technology & Business University, Beijing 100048, People’s Republic of China Y. Zhao Y. Deng Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, Jiangsu, People’s Republic of China
previous Bga-3. Higher glucose uptake rate, growth rate, and ethanol reuse rate were achieved in Bga-4 in shake-flask fermentation than those in Bga-3. Furthermore, the fed-batch fermentation in a 5-L bioreactor was performed without pH control, resulting in a final glucaric acid titer of 6.38 g/L. Conclusions Both the GA titer and biomass were enhanced along with the efficiency of ethanol reutilization in the presence of VHb. Moreover, the absence of base feeding for long-period fermentation reduced production cost, which is meaningful for industrial applications. Keywords Glucaric acid Saccharomyces cerevisiae Vitreoscilla hemoglobin pH-free fermentation
Introduction Saccharomyces cerevisiae, which is regarded as a eukaryotic model host for metabolic engineering of organic acid production, has excellent abilities for both stress resistance and low-pH fermentation with a relatively clear genetic background and convenient modification tools (Karathia et al. 2011). Unlike most bacteria such as Escherichia coli, it was unnecessary to control pH for S. cerevisiae to maintain a neutral environment by continuous consumption of base during organic acid fermentation; this could greatly
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decrease production cost. For dicarboxylate bio-synthesis, recombinant S. cerevisiae performed well in L-malic acid and succinate production, with high titers of 59 g/L and 43 g/L, respectively, making it a p
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