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High-expression and characterization of a novel serine protease from Ornithinibacillus caprae L9T with eco-friendly applications

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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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Authors and Affiliations

  1. Key Laboratory of Leather Chemistry and Engineering, Ministry of Education and College of Biomass Science and Engineering, Sichuan University, Chengdu, 610065, People’s Republic of China

    Xiaoguang Li, Zhe Xu, Guangyang Jiang, Longzhan Gan, Yongqiang Tian & Bi Shi

  2. Key Laboratory of Bio-Resources and Eco-Environment, Ministry of Education and College of Life Sciences, Sichuan University, Chengdu, 610065, People’s Republic of China

    Qian Zhang

Authors
  1. Xiaoguang Li
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  2. Qian Zhang
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  3. Zhe Xu
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  4. Guangyang Jiang
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  5. Longzhan Gan
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  6. Yongqiang Tian
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  7. Bi Shi
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Contributions

Xiaoguang 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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Correspondence to Yongqiang Tian or Bi Shi.

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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

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