Glycerol transporter 1 (Gt1) and zinc-regulated transporter 1 (Zrt1) function in different modes for zinc homeostasis in
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ORIGINAL RESEARCH PAPER
Glycerol transporter 1 (Gt1) and zinc-regulated transporter 1 (Zrt1) function in different modes for zinc homeostasis in Komagataella phaffii (Pichia pastoris) Yankun Yang . Yingyue Pan . Guoqiang Liu . Guibin Dong . Chunjun Zhan . Mattheos A. G. Koffas . Xiuxia Liu . Ye Li . Chunli Liu . Zhonghu Bai
Received: 1 February 2020 / Accepted: 7 July 2020 Ó Springer Nature B.V. 2020
Abstract Objectives To identify the zinc transport function of the membrane proteins Gt1 and Zrt1 in Komagataella phaffii (Pichia pastoris) and study their regulatory mode. Results Two membrane proteins that might have zinc transport function were found in K. phaffii. GT1 was known to encode a glycerol transporter belonging to the Major Facilitator Superfamily. ZRT1 was predicted to resemble the zinc transporter gene in Yankun Yang and Yingyue Pan contributed equally to this work.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02964-4) contains supplementary material, which is available to authorized users. Y. Yang (&) Y. Pan G. Liu G. Dong C. Zhan X. Liu Y. Li C. Liu Z. Bai Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Jiangnan University, Wuxi 214122, China e-mail: [email protected]
Saccharomyces cerevisiae. Consistent with the prediction, protein plasma-membrane localizations were confirmed by ultracentrifugation and confocal microscopy. Their zinc binding abilities were identified by ITC in vitro, and the impaired zinc uptake activity caused by their deficiencies was confirmed by zinc fluorescence quantification in vivo. Furthermore, zinc excess could turn the two channels off, while zinc deficiency induced their expressions. Gt1 could only function to maintain zinc homeostasis in glycerol, while the block of Gt1 function might lead to Zrt1 upregulation in glucose. Conclusions The zinc transport capabilities of Gt1 and Zrt1 were identified in vivo and in vitro. Their regulatory mode to maintain zinc homeostasis in K. phaffii is a new inspiration.
M. A. G. Koffas Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA
Y. Yang Y. Pan G. Liu G. Dong C. Zhan X. Liu Y. Li C. Liu Z. Bai National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China Y. Yang Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi 214122, China
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Biotechnol Lett
Keywords Komagataella phaffii Glycerol transporter 1 Zinc transporter Zinc homeostasis Protein trafficking Expression regulation Abbreviations MFS Major facilitator superfamily LZM Low-zinc medium ITC Isothermal titration calorimetry ZIF Zinc-induced facilitator ZIFL Zinc-induced-facilitator-like
Introduction Komagataella phaffii is a typical methylotrophic yeast widely used for heterologous protein expressions due to its strong- i
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