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Expression and Characterization of Surfactnt-Stable Calcium-Dependent Protease: a Potential Additive for Laundry Detergents

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Abstract

Extracellular protease SptA from the halophilic archaeon Natrinema sp. J7 was successfully expressed in Bacillus subtilis SCK6 using touchdown two-step PCR and plasmid multimer. Culture optimization was conducted to increase SptA protease activity 132-fold. It was shown that enzyme activity achieved the maximum activity at pH 8.0 and 55°C with casein as the substrate. SptA protease kept high stability and activity against many organic solvents and even maintained 60% activity in the presence of 8% NaCl. Meanwhile, SptA was redefined as a highly Ca2+-dependent subtilisin-like serine protease. Interestingly, SptA protease processed an autodigestion from 45 to 35 kDa in presence of Ca2+. Homology modeling illustrated the three binding sites of Ca2+ and the catalytic triad His231-Asp190-Ser384. In addition, SptA protease exhibited high compatibility with many surfactants and commercial detergents, showing 100% stability in the presence of commercial detergent Comfort. Washing performance analysis demonstrated remarkably high de-staining of the enzyme at 40°C for 20 min with a lower supplementation (500 U/mL). Accordingly, SptA protease could be considered as a promising bio-additive in the detergent industry.

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ACKNOWLEDGMENTS

This work was supported by the National Key Research and Development Program of China (2017YFB0308401).

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Authors and Affiliations

  1. College of Biomass Science and Engineering, Sichuan University, 610065, Chengdu, PR China

    H. Wang, S. Wang & Y. Tian

  2. Key Laboratory of Leather Chemistry and Engineering (Sichuan University), Ministry of Education, 610065, Chengdu, PR China

    H. Wang, C. Cai, L. Gan, S. Wang & Y. Tian

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  1. H. Wang
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  2. C. Cai
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  3. L. Gan
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Correspondence to Y. Tian.

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The authors declared that there are no competing interests. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Wang, H., Cai, C., Gan, L. et al. Expression and Characterization of Surfactnt-Stable Calcium-Dependent Protease: a Potential Additive for Laundry Detergents. Appl Biochem Microbiol 57, 481–492 (2021). https://doi.org/10.1134/S0003683821040165

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