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Production and characterization of milk-clotting enzyme from Bacillus amyloliquefaciens JNU002 by submerged fermentation

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Abstract

Milk-clotting enzymes produced by microorganisms have been developed to replace calf rennet, yet the enzymatic level and the ratio of milk-clotting activity to proteolytic activity still need further improvement. This work described a strain Bacillus amyloliquefaciens JNU002 that screened from wheat bran that has a promising characterization. After optimization, B. amyloliquefaciens JNU002 showed a high milk-clotting activity (4969 SU/mL) and low proteolytic activity (4.02 U/mL) at 48 h with an inoculum size of 0.2% (v/v) at initial pH 6.0 in 15-L bioreactor. After purification, the purified enzyme gave a single protein band on SDS–PAGE, corresponding to 28 kDa. The purified enzyme showed a high ratio (2,575) of milk-clotting activity to proteolytic activity at 35 °C, and the ratio would be even higher (22,992) at 70 °C. The milk-clotting reaction and proteolytic reaction were prevented at 75 °C. This enzyme was stable at pH 4–6 and below 40 °C, and this was convenient for storage and transportation.

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Acknowledgments

This study is supported by the National High Technology Research and Development Program of China (No. 2011AA100905) and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, People’s Republic of China

    Zhongyang Ding, Wangfei Wang, Boda Wang, Anran Ouyang, Siwei ** Liu, Mingliang Ding, Liang Zhang & Guiyang Shi

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  1. Zhongyang Ding
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  2. Wangfei Wang
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Correspondence to Zhongyang Ding.

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Ding, Z., Wang, W., Wang, B. et al. Production and characterization of milk-clotting enzyme from Bacillus amyloliquefaciens JNU002 by submerged fermentation. Eur Food Res Technol 234, 415–421 (2012). https://doi.org/10.1007/s00217-011-1650-2

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  • DOI: https://doi.org/10.1007/s00217-011-1650-2

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