Biotechnology Bulletin ›› 2020, Vol. 36 ›› Issue (12): 82-90.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0375

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Cloning and Enzymatic Identification of Thermo-tolerant and Endotype Alginate Lyase Gene from Mangrovibacterium sp. SH-52

LI Wei-na1(), SHEN Dong-ling2, ZHANG Yu-xing1, LIU Xue-tong1, IRBIS Chagan2()   

  1. 1. School of Medicine,Xijing University,Xi’an 710000;
    2. Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500
  • Received:2020-04-06 Online:2020-12-26 Published:2020-12-22
  • Contact: IRBIS Chagan E-mail:liweina502361483@163.com;chaganirbis2019@126.com

Abstract:

The objective of this work is to investigate the alginate lyases isolated from anaerobic alginolytic bacteria,to characterize the novel alginate lyase,thus to provide a theoretical basis for the enriching industrial application of alginate lyase. A gene SHA-I encoding an alginate lyase was cloned from an anaerobic bacterium Mangrovibacterium sp. SH-52,sequenced and expressed in Escherichia coli. Its expressed enzyme was purified and the enzymatic characteristics were analyzed. The alginate lyase SHA-I was composed of 362 amino acids with molecular weight of approximately 41 kD. SHA-I belonged to polysaccharide lyase(PL)family 7 and showed 83% amino acid identity with alginate lyase from Lewinella agarilytica. The optimal pH for SHA-I activity was 7.0. SHA-I demonstrated the highest activity at 50℃and remained approximate 60% of the highest activity at 60℃,and 40% at 80℃,indicating that SHA-I was a thermo-tolerant alginate lyase. SHA-I presented substrate specificity to polyG(polymannuronic acid). It degraded polyG blocks to alginate oligosaccharides as the main products.

Key words: anaerobic alginolytic bacteria, Mangrovibacterium sp. SH-52, heterologous expression, endotype alginate lyase, thermo-tolerant