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Characterization and Optimization of Amylase Production in WangLB, a High Amylase-Producing Strain of Bacillus

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Abstract

The costs of amylase represent ca. 24 % of the expenditures in the starch industry and an increase in amylase production and/or activity will greatly cut down on production costs. In the present study, we obtained a high amylase-producing strain of bacteria, WangLB, and identified it as a member of the Bacillus genus based on 16S rDNA analysis. The fermentation conditions for amylase production in the strain were optimized, and the maximum amylase activity we obtained was 26,670 ± 1390 U/mL, under the optimized conditions of 48-h incubation in liquid starch medium, 35 °C, pH 10, 1 % v/v inoculum concentration, 20 g/L starch concentration, and 0.1 % w/v peptone. The influences of 16 small organic inducers on amylase production were tested, and the results showed that 20 mmol/L alanine greatly enhanced amylase production to 290 % of the baseline level. We also conducted an amylase enzymology analysis. The molecular weight of the amylase was 55 kD, determined by SDS-PAGE. The optimum temperature and pH for the amylase were 55 °C and pH 9, respectively. The enzyme also showed high activity over a wide range of temperatures (50–85 °C) and pH values (3–10), and the activity of the amylase was Ca2+ independent. The kinetic parameters K m and V max were 0.37 ± 0.02 mg/mL and 233 U/mg, respectively. Finally, the amylase was applied to the hydrolysis of five different brands of starch. It was found that the hydrolyzability of the substrate by amylase increased along with starch solubility.

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Acknowledgments

This work was financially supported by the Natural Sciences and Engineering Research Council of Canada (NSERC)—Research Development Fund (RDF) to W. Q. The authors would like to thank Dr. Md. Shafiqur Rahman for his assistance and advice with the experiment in the manuscript.

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Authors and Affiliations

  1. Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China

    Shihui Wang & Yun Liu

  2. Department of Biology, Lakehead University, 955 Oliver Road, Thunder Bay, Ontario, P7B 5E1, Canada

    Shihui Wang, Jenasia Jeyaseelan & Wensheng Qin

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  1. Shihui Wang
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  2. Jenasia Jeyaseelan
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  3. Yun Liu
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  4. Wensheng Qin
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Correspondence to Wensheng Qin.

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Wang, S., Jeyaseelan, J., Liu, Y. et al. Characterization and Optimization of Amylase Production in WangLB, a High Amylase-Producing Strain of Bacillus . Appl Biochem Biotechnol 180, 136–151 (2016). https://doi.org/10.1007/s12010-016-2089-5

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  • DOI: https://doi.org/10.1007/s12010-016-2089-5

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