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Cloning and overexpression of aprE3-17 encoding the major fibrinolytic protease of Bacillus licheniformis CH 3-17

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Abstract

Bacillus licheniformis (B. licheniformis) CH3-17, an isolate from cheonggukjang, a traditional Korean fermented soyfood, secretes several fibrinolytic enzymes into the culture medium, showing strong fibrinolytic activity. A gene homologous to aprE of Bacillus subtilis (B. subtilis), aprE3-17, was cloned by PCR. DNA sequencing showed that aprE3-17 encodes a prepro-type serine protease consisting of 382 amino acids. The mature enzyme was 27 kDa in size. The aprE3-17 gene was overexpressed in B. subtilis WB600 using pHY300PLK, an Escherichia coli (E. coli)-Bacillus shuttle vector, and the 27 kDa enzyme was purified from the culture supernatant. The optimum pH for activity was 6.0. Purified enzyme quickly degraded the Aα and Bβ chains of fibrinogen but could not degrade the γ-chain.

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Authors and Affiliations

  1. Division of Applied Life Science (BK21), Graduate School, Gyeongsang National University, Jinju, 660-701, Korea

    Hyeon-Deok Jo & Jeong Hwan Kim

  2. Institute of Agriculture & Life Science, Gyeongsang National University, Jinju, 660-701, Korea

    Gun-Hee Kwon & Jeong Hwan Kim

  3. Department of Food Science and Nutrition, Catholic University of Daegu, Gyeongsan, 712-702, Korea

    Jae-Yong Park

  4. Department of Microbiology, Pusan National University, Busan, 609-735, Korea

    Jaeho Cha

  5. School of Food and Life Science, Inje University, Gimhae, 621-749, Korea

    Young-Sun Song

Authors
  1. Hyeon-Deok Jo
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  2. Gun-Hee Kwon
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  3. Jae-Yong Park
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  4. Jaeho Cha
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  5. Young-Sun Song
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  6. Jeong Hwan Kim
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Correspondence to Jeong Hwan Kim.

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Jo, HD., Kwon, GH., Park, JY. et al. Cloning and overexpression of aprE3-17 encoding the major fibrinolytic protease of Bacillus licheniformis CH 3-17. Biotechnol Bioproc E 16, 352–359 (2011). https://doi.org/10.1007/s12257-010-0328-0

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  • DOI: https://doi.org/10.1007/s12257-010-0328-0

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