Abstract
In the current work, a novel thermophilic serine protease gene (P3862) from Ornithinibacillus caprae L9T was functionally expressed in Bacillus subtilis SCK6. The monomeric enzyme of about 29 kDa was purified to homogeneity with 43.91% of recovery and 2.81-folds of purification. Characterization of the purified protease revealed the optimum activity at pH 7 and 65 °C. The protease exhibited excellent activity and stability in the presence of Na+, Mg2+, Ca2+, ethanediol, n-hexane, Tween-20, Tween-80 and Triton X-100. P3862 displayed favorable caseinolytic activity, moderate keratinolytic activity but no collagenolytic activity. Besides, the homology model of P3862 possessed a globular configuration and characteristic of α/β hydrolase fold, and displayed stable interactions with casein, glycoprotein and keratin rather than collagen. Moreover, the crude enzyme could completely dehair goatskin within 6 h, resulting in decrease in BOD5, COD and TSS loads by 72.86, 74.07, and 73.79%, respectively, as compared with Na2S treatment. Biocatalytic applications revealed that it could effectively remove egg-stains from fabrics at 37 °C for 30 min with low supplementation (300 U/mL), and was able to degrade the feathers of duck and chicken. Overall, these outstanding properties make P3862 valuable in the development of environmentally friendly biotechnologies
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Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- BGSC:
-
Bacillus Genetic Stock Center;
- BOD:
-
biological oxygen demand;
- COD:
-
chemical oxygen demand;
- EDTA:
-
ethylenediaminetetraacetic acid;
- EGTA:
-
ethylene glycol-bis (β-aminoethyl ether)-N,N,N’,N’-tetraacetic acid;
- HE:
-
hematoxylin and eosin;
- KCTC:
-
Kalona Cooperative Technology Company;
- β-ME:
-
β-mercaptoethanol;
- PCR:
-
polymerase chain reaction;
- PMSF:
-
phenylmethanesulfonyl fluoride;
- SD:
-
standard deviation;
- SDS-PAGE:
-
sodium dodecyl sulfate polyacrylamide gel electrophoresis;
- SEM:
-
scanning electron microscope;
- TSS:
-
total suspended solids.
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Funding
This work was supported by the National Key Research and Development Program of China (2017YFB0308401) and the Key Program of Sichuan Science and Technology Project (2020JDPT0018).
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**aoguang Li: methodology, formal analysis, writing—original draft. Qian Zhang: formal analysis, writing—original draft. Zhe Xu: Data curation, investigation. Guangyang Jiang: data curation. Longzhan Gan: formal analysis. Yongqiang Tian: conceptualization, supervision, resources. Bi Shi: supervision, conceptualization.
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Highlights
• A novel protease named P3862 was heterologously expressed in Bacillus subtilis SCK6.
• The monomeric enzyme of about 29 kDa showed high stability to nonionic surfactants.
• The model interacts well with casein, glycoprotein and keratin rather than collagen.
• The crude enzyme completely dehaired goatskin with notably reduced pollutant content.
• It exhibited promising result in the degradation of feathers and washing of stains.
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Li, X., Zhang, Q., Xu, Z. et al. High-expression and characterization of a novel serine protease from Ornithinibacillus caprae L9T with eco-friendly applications. Environ Sci Pollut Res 29, 35996–36012 (2022). https://doi.org/10.1007/s11356-021-17495-2
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DOI: https://doi.org/10.1007/s11356-021-17495-2