Abstract
As an approach to the exploration of cold-active enzymes, in this study, we isolated a cold-active protease produced by psychrotrophic bacteria from glacial soils of Thajwas Glacier, Himalayas. The isolated strain BO1, identified as Bacillus pumilus, grew well within a temperature range of 4–30 °C. After its qualitative and quantitative screening, the cold-active protease (Apr-BO1) was purified. The Apr-BO1 had a molecular mass of 38 kDa and showed maximum (37.02 U/mg) specific activity at 20 °C, with casein as substrate. It was stable and active between the temperature range of 5–35 °C and pH 6.0–12.0, with an optimum temperature of 20 °C at pH 9.0. The Apr-BO1 had low Km value of 1.0 mg/ml and Vmax 10.0 µmol/ml/min. Moreover, it displayed better tolerance to organic solvents, surfactants, metal ions and reducing agents than most alkaline proteases. The results exhibited that it effectively removed the stains even in a cold wash and could be considered a decent detergent additive. Furthermore, through protein modelling, the structure of this protease was generated from template, subtilisin E of Bacillus subtilis (PDB ID: 3WHI), and different methods checked its quality. For the first time, this study reported the protein sequence for psychrotrophic Apr-BO1 and brought forth its novelty among other cold-active proteases.
Highlights
As an approach to the exploration of cold-active enzymes, in this study, we isolated a cold-active protease produced by psychrotrophic bacteria from glacial soils of Thajwas Glacier, Himalayas
While the contribution of bacterial alkaline proteases to total protease market share stands at 60%10–12, the data related to cold-active alkaline proteases from psychrotrophs is essentially missing[4]
The dearth of novel alkaline cold-active proteases characterized by being active at low temperatures, stable in the presence of metal ions, surfactants, organic solvents and oxidizing agents still exists
Summary
As an approach to the exploration of cold-active enzymes, in this study, we isolated a cold-active protease produced by psychrotrophic bacteria from glacial soils of Thajwas Glacier, Himalayas. The prime significance of these bacteria stems from their ability to survive and retain functionality in cold temperatures and grow optimally even at warm temperatures[3] These bacteria offer an immense natural resource of cold-active enzymes that function at low temperatures (10–20 °C). The dearth of novel alkaline cold-active proteases characterized by being active at low temperatures, stable in the presence of metal ions, surfactants, organic solvents and oxidizing agents still exists. The main aim of the present study was to purify and characterize serine alkaline protease from a psychrotrophic bacterium that can catalyze reactions at low temperature and be used as an industrial additive. This study successfully characterized and purified cold-active serine alkaline protease from psychrotrophic Bacillus pumilus BO1 isolated from western Himalayan glacial soil
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