Screening and characterization of proteases produced by deep-sea cold seep bacteria -- Journal of Oceanology and Limnology,2022,40(2):678-689

	Cite this paper:
	Chenchen GUO, Chaomin SUN, Shimei WU. Screening and characterization of proteases produced by deep-sea cold seep bacteria[J]. Journal of Oceanology and Limnology, 2022, 40(2): 678-689

Screening and characterization of proteases produced by deep-sea cold seep bacteria

Chenchen GUO¹, Chaomin SUN², Shimei WU¹

1 College of Life Sciences, Qingdao University, Qingdao 266071, China;
2 CAS Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China

Abstract:

Fifty protease-producing strains were screened from sediment of deep-sea cold seep, and divided into four different categories:Bacillus, Pseudoalteromonas, Vibrio, and Alteromonas according to the sequences of 16s rRNA. Their abilities to produce protease, amylase, and lipase were determined, and a Bacillus strain gcc-1 displayed very strong alkaline protease activity and stability under different thermal and acidic conditions. The purification of the protease produced by strain gcc-1 was carried out by precipitation with ammonium sulfate, and sequentially chromatographed by anion exchange column and gel filtration. The purified protease showed a single band at the molecular weight of 28 kDa by SDS-PAGE. The characterization results show that the purified protease exhibited a considerable activity and stability in a wide thermal range of 10-80℃ and a wide acidic range of pH 6.5-11.5, and displayed highest activity at 40℃ and pH 8.5. Notably, the protease still maintained high activity even at low to 10℃. Furthermore, the protease exhibited good stability in presence of different surfactants, organic solvents, oxidizing agent H₂O₂, and commercial detergents. Therefore, the protease produced by gcc-1 is a cold active and high stable enzyme, and has a promising potential in laundry detergent as an additive.

Key words: deep-sea|Bacillus|alkaline protease|purification|characterization

Received: 2020-11-20 Revised:

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Articles by Chenchen GUO

Articles by Chaomin SUN

Articles by Shimei WU

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