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A novel cold-adapted lipase from Acinetobacter sp. XMZ-26: gene cloning and characterisation

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Abstract

Acinetobacter sp. XMZ-26 (ACCC 05422) was isolated from soil samples obtained from glaciers in Xinjiang Province, China. The partial nucleotide sequence of a lipase gene was obtained by touchdown PCR using degenerate primers designed based on the conserved domains of cold-adapted lipases. Subsequently, a complete gene sequence encoding a 317 amino acid polypeptide was identified. Our novel lipase gene, lipA, was overexpressed in Escherichia coli. The recombinant protein (LipA) was purified by Ni-affinity chromatography, and then deeply characterised. The LipA resulted to hydrolyse pNP esters of fatty acids with acyl chain length from C2 to C16, and the preferred substrate was pNP octanoate showing a kcat = 560.52 ± 28.32 s−1, Km = 0.075 ± 0.008 mM, and a kcat/Km = 7,377.29 ± 118.88 s−1 mM−1. Maximal LipA activity was observed at a temperature of 15°C and pH 10.0 using pNP decanoate as substrate. That LipA peaked at such a low temperature and remained most activity between 5°C and 35°C indicated that it was a cold-adapted enzyme. Remarkably, this lipase retained much of its activity in the presence of commercial detergents and organic solvents, including Ninol, Triton X-100, methanol, PEG-600, and DMSO. This cold-adapted lipase may find applications in the detergent industry and organic synthesis.

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Authors and Affiliations

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081, People’s Republic of China

    Xiaomei Zheng, Xiaoyu Chu, Wei Zhang, Ningfeng Wu & Yunliu Fan

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  1. Xiaomei Zheng
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  2. Xiaoyu Chu
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  3. Wei Zhang
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  4. Ningfeng Wu
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  5. Yunliu Fan
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Correspondence to Ningfeng Wu.

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Zheng, X., Chu, X., Zhang, W. et al. A novel cold-adapted lipase from Acinetobacter sp. XMZ-26: gene cloning and characterisation. Appl Microbiol Biotechnol 90, 971–980 (2011). https://doi.org/10.1007/s00253-011-3154-1

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  • DOI: https://doi.org/10.1007/s00253-011-3154-1

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