Functional analysis of PGI1 and ZWF1 in thermotolerant yeast Kluyveromyces marxianus

Zhang, Biao; Ren, Lili; Zeng, Shuai; Zhang, Siyang; Xu, Dayong; Zeng, Xin; Li, Feng

doi:10.1007/s00253-020-10808-4

Applied microbial and cell physiology
Published: 10 August 2020

Functional analysis of PGI1 and ZWF1 in thermotolerant yeast Kluyveromyces marxianus

Biao Zhang ORCID: orcid.org/0000-0002-9627-7789¹^na1,
Lili Ren¹^na1,
Shuai Zeng¹,
Siyang Zhang¹,
Dayong Xu¹,
Xin Zeng¹ &
Feng Li¹

Applied Microbiology and Biotechnology volume 104, pages 7991–8006 (2020)Cite this article

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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 co-utilized 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.

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Funding

This study was funded by the Natural Science Foundation of Anhui Province (grant no. 1908085QC125), the University Natural Science Research Project of Anhui Province (grant no. KJ2018A0399), and the Innovation Team of Scientific Research Platform of Anhui Province (grant no. KJ2015TD001).

Author information

Biao Zhang and Lili Ren contributed equally to this work.

Affiliations

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
Biao Zhang, Lili Ren, Shuai Zeng, Siyang Zhang, Dayong Xu, Xin Zeng & Feng Li

Authors

Biao Zhang
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Lili Ren
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Shuai Zeng
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Siyang Zhang
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Dayong Xu
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Xin Zeng
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Feng Li
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Contributions

B. Z. and LL. R. conceived and designed the research. B. Z., LL, R., S. Z., and SY. Z. conducted the experiments. DY. X. and X. Z. contributed the new reagents or analytical tools. B. Z. and LL. R. analyzed the data. B. Z., LL. R., and F. L. wrote the manuscript. B. Z., LL. R., DY. X., X. Z., and F. L. revised the manuscript. All authors read and approved the manuscript.

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Correspondence to Biao Zhang or Feng Li.

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Zhang, B., Ren, L., Zeng, S. et al. Functional analysis of PGI1 and ZWF1 in thermotolerant yeast Kluyveromyces marxianus. Appl Microbiol Biotechnol 104, 7991–8006 (2020). https://doi.org/10.1007/s00253-020-10808-4

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Received: 30 April 2020
Revised: 06 July 2020
Accepted: 02 August 2020
Published: 10 August 2020
Issue Date: September 2020
DOI: https://doi.org/10.1007/s00253-020-10808-4

Keywords

Kluyveromyces marxianus
PGI1
ZWF1
NADP⁺
NADPH