Functional analysis of PGI1 and ZWF1 in thermotolerant yeast Kluyveromyces marxianus
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APPLIED MICROBIAL AND CELL PHYSIOLOGY
Functional analysis of PGI1 and ZWF1 in thermotolerant yeast Kluyveromyces marxianus Biao Zhang 1
&
Lili Ren 1 & Shuai Zeng 1 & Siyang Zhang 1 & Dayong Xu 1 & Xin Zeng 1 & Feng Li 1
Received: 30 April 2020 / Revised: 6 July 2020 / Accepted: 2 August 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Glycolysis and the pentose phosphate pathway (PPP) are two basic metabolic pathways that are simultaneously present in yeasts. As the main pathway in most species, the glycolysis provides ATP and NADH for cell metabolism while PPP, as a complementary pathway, supplies NADPH. In this study, the performance of Kluyveromyces marxianus using glycolysis or PPP were studied through the disruption of PGI1 or ZWF1 gene, respectively. K. marxianus using glycolysis as the only pathway showed higher ethanol production ability than that of the Kluyveromyces lactis zwf1Δ mutant; K. marxianus using only PPP showed more robustness than that of Saccharomyces cerevisiae pgi1Δ mutant. Additionally, K. marxianus pgi1Δ strain accumulated much more intracellular NADPH than the wild type strain and coutilized glucose and xylose more effectively. These findings suggest that phosphoglucose isomerase participates in the regulation of the repression of glucose on xylose utilization in K. marxianus. The NADPH/NADP+ ratio, dependent on the activity of the PPP, regulated the expression of multiple genes related to NADPH metabolism in K. marxianus (including NDE1, NDE2, GLR1, and GDP1). Since K. marxianus is considered a promising host in industrial biotechnology to produce renewable chemicals from plant biomass feedstocks, our research showed the potential of the thermotolerant K. marxianus to produce NADP(H)-dependent chemical synthesis from multiple feedstocks. Key points • The function of PGI1 and ZWF1 in K. marxianus has been analyzed in this study. • K. marxianus zwf1Δ strain produced ethanol but with decreased productivity. • K. marxianus pgi1Δ strain grew with glucose and accumulated NADPH. • K. marxianus pgi1Δ strain released glucose repression on xylose utilization. Keywords Kluyveromyces marxianus . PGI1 . ZWF1 . NADP+ . NADPH
Introduction Biao Zhang and Lili Ren contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10808-4) contains supplementary material, which is available to authorized users. * Biao Zhang [email protected] * Feng Li [email protected] 1
Anhui Province Key Laboratory of Pollutant Sensitive Materials and Environmental Remediation, School of Life Sciences, Huaibei Normal University, Huaibei, Anhui 235000, People’s Republic of China
The pentose phosphate pathway (PPP) branches from glycolysis at the level of glucose-6-phosphate (G6P) (Fig. 1). PPP generates the major source of NADPH required for the neutralization of reactive oxygen species (ROS), reductive biosynthetic reactions, and the production of metabolic intermediates (Saliola et al. 2007). As a highly conserved
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