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A cold-active β-glucosidase (Bgl1C) from a sea bacteria Exiguobacterium oxidotolerans A011

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Abstract

By constructing a genomic library, a new gene encoding β-glucosidase (Bgl1C) was cloned from Exiguobacterium oxidotolerans A011, which was isolated from deep sea mud. The putative β-glucosidase gene consisted of an open reading frame (ORF) of 1,347 nucleotides, and encoded a protein of 448 amino acids with a predicted molecular weight of 51.6 kDa. Bgl1C belonged to the glycoside hydrolase family 1, and the deduced amino acid sequence displayed the highest identity (68%) to the β-glucosidase from Bacillus coahuilensis m4-4. Optimal conditions for activity were pH 7 and a temperature of 35°C and Bgl1C was stable in buffers ranging from pH 6.6 to 9. The specific activity, K m, and V max for the substrate p-nitrophenyl-β-d-glucopyranoside were 41 U mg−1, 1.72 mg ml−1 and 0.45 μg ml−1 s−1, respectively. Na+, Ca2+, EDTA and β-mercaptoethanol had no effect on the activity, while Hg2+, Cu2+, Co2+ strongly inhibited it. It is noteworthy that Bgl1C is a cold active enzyme that retains about 61% of its maximum activity at 10°C. Structural model of Bgl1C revealed that some amino acids (glycine, alanine, serine, valine) concerned with plasticity and flexibility were located around the active sites, this may contributed to the cold adaption of Bgl1C. These favorable features make Bgl1C a potential candidate for various industrial applications.

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Acknowledgments

We thank Dr. Qifa Zhang for many valuable suggestions. This work was supported by grants from the National Natural Sciences Foundation of China (30770021 and 30570057) and the 111 project (B07041).

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Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Shuilian Chen & Ziduo Liu

  2. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Yuzhi Hong

  3. Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State of Oceanic Administration, Xiamen, 361005, People’s Republic of China

    Zongze Shao

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  1. Shuilian Chen
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  2. Yuzhi Hong
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  3. Zongze Shao
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  4. Ziduo Liu
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Correspondence to Ziduo Liu.

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Chen, S., Hong, Y., Shao, Z. et al. A cold-active β-glucosidase (Bgl1C) from a sea bacteria Exiguobacterium oxidotolerans A011. World J Microbiol Biotechnol 26, 1427–1435 (2010). https://doi.org/10.1007/s11274-010-0317-7

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  • DOI: https://doi.org/10.1007/s11274-010-0317-7

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