Purification, characterization, and gene cloning of a cold-adapted thermolysin-like protease from Halobacillus sp SCSIO 20089
作者:
[Yang, Jian
; Li, Jie
; Mai, Zhimao
; Tian, Xinpeng
; Zhang, Si] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Marine Bioresources Sustainable Utilizat, Guangzhou 510301, Guangdong, Peoples R China
; [Yang, Jian
; Mai, Zhimao
; Zhang, Si] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
Marine sediment is a distinctive habitat of cold enzyme producing bacteria. A protease producing strain Halobacillus sp. SCSIO 20089 was isolated from a marine sediment of South China Sea. Using chromatographic techniques, the extracellular protease was purified to homogeneity from the culture supernatant. The purified protease exhibited maximal activity at 30 degrees C, pH 8.0, and remained more than 20% of its activity at 0 degrees C. Its activation energy was calculated to be 34.4 kJ/mol, suggesting it is a cold-adapted protease. Based on the N-terminal amino acid sequence of the purified enzyme, full gene encoding the enzyme was obtained by combination of degenerate primer PCR and hiTAIL-PCR. The deduced amino acid sequence showed 57% and 52% identity with mesothermal and thermophilic protease in thermolysin family respectively. All these indicate the enzyme is a unique cold-active thermolysin-like protease with potential in both basic research and industrial application areas. (C) 2012, The Society for Biotechnology, japan. All rights reserved.
[Yang, Jian; Li, Jie; Mai, Zhimao; Tian, Xinpeng; Zhang, Si] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Marine Bioresources Sustainable Utilizat, Guangzhou 510301, Guangdong, Peoples R China; [Yang, Jian; Mai, Zhimao; Zhang, Si] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China.Purification, characterization, and gene cloning of a cold-adapted thermolysin-like protease from Halobacillus sp SCSIO 20089,JOURNAL OF BIOSCIENCE AND BIOENGINEERING,2013,115(6):628-632
