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Cloning and expression analysis of tps, and cryopreservation research of trehalose from Antarctic strain Pseudozyma sp.

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Abstract

Trehalose is a non-reducing disaccharide sugar that widely exists in a variety of organisms, such as bacteria and eukaryotes except the vertebrates. It plays an important role in a number of critical metabolic functions especially in response to stressful environmental conditions. However, the biosynthetic pathways of trehalose in cold-adapted yeast and its responses to temperature and salinity changes remain little understood. In this study, the genome of Antarctic-isolated Pseudozyma sp. NJ7 was generated from which we identified the gene coding for trehalose phosphate synthase (TPS1) and trehalose phosphate phosphatase (TPS2), the two enzymes most critical for trehalose production. The whole draft genome length of Pseudozyma sp. NJ7 was 18,021,233 bp, and encoded at least 34 rRNA operons and 72 tRNAs. The open reading frame of tps1 contained 1827 nucleotide encoding 608 amino acids with a molecular weight of 67.64 kDa, and an isoelectric point of 5.54, while tps2 contained 3948 nucleotide encoding 1315 amino acids with a molecular weight of 144.47 kDa and an isoelectric point of 6.36. The TPS1 and TPS2 protein sequences were highly homologous to Moesziomyces antarcticus T-34, but TPS2 had obvious specificity and differently with others which suggest species specificity and different evolutionary history. Expression level of tps1 gene was strongly influenced by temperature and high salinity. In addition, addition of 0.5% trehalose preserved yeast cells in the short term but was not effective for cryopreservation for more than 5 days, but still suggesting that exogenous trehalose could indeed significantly improve the survival of yeast cells under freezing conditions. Our results provided new insights on the molecular basis of cold adaptations of Antarctic Pseudozyma sp., and also generated new information on the roles trehalose play in yeast tolerance to extreme conditions in the extreme Antarctic environments.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (No. 41576187), the Natural Science Foundation of China-Shandong Joint Fund (No. U1606403), the Polar Strategic Foundation of China (No. 20150303), the Shandong Provincial Natural Science Foundation, China (No. ZR2017QD008), the Public Science and Technology Research Funds Projects of Ocean (No. 201405015), the Key Research and Development Program of Shandong Province (No. 2016ZDJS06A03), the Scientific and Technological Innovation Project Financially Supported by Qingdao National Laboratory for Marine Science and Technology and the Science (No. 2015ASKJ02), the Open Research Fund of State Key Laboratory of Biological Fermentation Engineering of Beer (No. K2014002), the Technology Planning Project of Shandong Province (No. 2014GHY115003), and Qingdao Entrepreneurship and Innovation Pioneers Program (No. 15-10-3-15-(44)-zch). We thank MogoEdit Co. for its linguistic assistance during the preparation of this manuscript.

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  1. Hua Yin and Yibin Wang contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Biological Fermentation Engineering of Beer, Tsingtao Brewery Co. Ltd, Qingdao, 266061, China

    Hua Yin & Lei Xing

  2. The First Institute of Oceanography, State Oceanic Administration, Qingdao, 266061, China

    Yibin Wang, Yingying He, Shuai Wang, Xiaoqing Qi, Zhou Zheng & Jinlai Miao

  3. Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266235, China

    Yibin Wang, Zhou Zheng & Jinlai Miao

  4. Clinical Laboratory, Qingdao Hiser Medical Center, Qingdao, 266033, China

    Xiufang Zhang

  5. Marine and Fisheries Monitoring Center of Sanya, Sanya, 572000, China

    Shuai Wang

  6. Institute of Deep-Sea Science and Engineering, Chinese Academy of Sciences, Sanya, 572000, China

    Xiaoqing Qi

  7. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China

    Jian Lu

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  1. Hua Yin
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Correspondence to Jinlai Miao.

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Yin, H., Wang, Y., He, Y. et al. Cloning and expression analysis of tps, and cryopreservation research of trehalose from Antarctic strain Pseudozyma sp.. 3 Biotech 7, 343 (2017). https://doi.org/10.1007/s13205-017-0983-3

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