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Isolation and characterization of a cold-active, detergent-stable protease from Serratia sp. TGS1.

Sahib ZadaMohsin KhanWasim SajjadMuhammad RafiqWasim SajjadZheng Su
Published in: Journal of basic microbiology (2023)
Psychrophiles are cold-adapted microorganisms living in cold regions and are known to generate cold-active enzymes such as proteases, lipases, and peptidases. These types of enzymes are a major part of the market of the food and textile sector. This study aimed to isolate and characterize the cold-active and detergent-stable, extracellular protease from psychotrophic bacteria Serratia sp. TGS1 (OQ654005). Protease was purified by gel permeation chromatography using Sephadex G-75. The specific activity of the purified protease was 250 U/mg at 15°C, with a purification fold of 5.68 and a percentage yield of 60%. The cold active protease was stable within a temperature range of 5-30°C and a pH range of 6-10. Ca +2 and Mg +2 enhanced its activity while chelators like ethylenediaminetetraacetic acid inhibited cold active protease, showing it as metalloprotease in nature. The enzyme was sensitive to Cu +2 , Zn +2 , and Hg +2 , and the proteolytic activity decreased upon treatment with heavy metals. The molecular weight of the protease was estimated to be 47 kDa using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Proteins within a specific range of molecular weight possess desirable properties for industrial enzyme use. By working on a specific range, the researchers intended to examine an enzyme to examine its specific characteristics. The purified protease showed high stability to detergents like SDS, Tween 20, Tween 60, and Triton X. The maximum velocity V max and K m values were 59.90 mg/min/mL and 1.53 mg/mL, respectively. The obtained protease exhibited an interesting activity at a broad range of pH (6-10) and stability at low temperatures (5-30°C) and detergents. Such enzymatic features of versatile and potent cold-active enzymes enhance their industrial applications to meet food, dairy, and laundry requirements.
Keyphrases
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  • wastewater treatment
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