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Cloning of Exo-β-1,3-glucanase Gene from a Marine Yeast Williopsis saturnus and Its Overexpression in Yarrowia lipolytica

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Abstract

The exo-β-1,3-glucanase structural gene (WsEXG1 gene, accession number: FJ875997.2) was isolated from both the genomic DNA and cDNA of the marine yeast Williopsis saturnus WC91-2 by inverse PCR and RT-PCR. An open reading frame of 1,254 bp encoding a 417 amino acid protein (isoelectric point: 4.5) with calculated molecular weight of 46.2 kDa was characterized. The promoter of the gene (intronless) was located from −28 to −77 and had one TATA box while its terminator contained the sequence AAGAACAATAAACAA from +1,386 to +1,401. The protein had the Family 5 glycoside hydrolase signature IGLELLNEPL and a fragment with the sequence of NLCGEWSAA, where the Glu-310 (E) was considered to be the catalytic nucleophile. The WsEXG1 gene was overexpressed in Yarrowia lipolytica Po1h and the recombinant WsEXG1 was purified and characterized. The molecular weight of the purified rWsEXG1 was 46.0 kDa. The optimal pH and temperature of the purified rWsEXG1 were 5.0°C and 40°C, respectively. The purified rWsEXG1 had high exo-β-1,3-glucanase activity. Therefore, the recombinant β-1,3-glucanase may have highly potential applications in food and pharmaceutical industries.

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Acknowledgment

This research was supported by the National Natural Science Foundation of China, grant number 30670058.

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Authors and Affiliations

  1. UNESCO Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No.5, Qingdao, China

    Ying Peng, Guang-Lei Liu, Xin-Jun Yu, Xiang-Hong Wang, Li Jing & Zhen-Ming Chi

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  1. Ying Peng
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  2. Guang-Lei Liu
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  3. Xin-Jun Yu
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  4. Xiang-Hong Wang
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  5. Li Jing
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  6. Zhen-Ming Chi
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Correspondence to Zhen-Ming Chi.

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Peng, Y., Liu, GL., Yu, XJ. et al. Cloning of Exo-β-1,3-glucanase Gene from a Marine Yeast Williopsis saturnus and Its Overexpression in Yarrowia lipolytica . Mar Biotechnol 13, 193–204 (2011). https://doi.org/10.1007/s10126-010-9281-3

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  • DOI: https://doi.org/10.1007/s10126-010-9281-3

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