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In Silico Study of Alkaline Serine Protease and Production Optimization in Bacillus sp. Khoz1 Closed Bacillus safensis Isolated from Honey

  • Babak Elyasi Far
  • Ahmad Yari Khosroushahi
  • Azita DilmaghaniEmail author
Article
  • 12 Downloads

Abstract

Protease is a kind of enzyme with different applications in medicine and industry. The limitation of the enzymes in industry is lack of tolerance for harsh conditions (high temperature, pH, and low water condition). This study intends to evaluate in silico study and screen the alkaline serine protease producing bacteria from honey and optimize the production of low water active alkaline serine protease. Plackett–Burman method was applied to improve effective factors on protease production, including pH, inoculum size, temperature, time, soybean protein, KH2PO4, MgSO4, CaCl2, NaCl, Glycerin and glucose concentration. Further, the Box–Behnken method was used to optimize the significant level of selected factors. Besides, in silico study was utilized to create the enzyme structure and investigate the stability in harsh conditions. The results showed that the highest protease production belongs to Bacillus sp. Khoz1 closed Bacillus safensis, Also temperature, glucose and soybean protein concentration were three significant factors for protease production and the optimized level for them were 35 °C, 0.5 g/l, and 38.32 g/l, respectively. The microorganism was able to produce protease until 98.36 U/ml and it was 98.68% similar to B. safensis. The stability of microbial alkaline serine protease was also determined in high pH and temperature conditions. The best stable condition for isolated protease was pH 9 and temperature at 50 °C. The in silico data showed that this protease has similar 3D structure to Bacillus subtilis Subtilisin E and highly charge amino acids on its surface were that caused this enzyme still activate and stable in low water condition.

Keywords

Low waster activity bacteria Honey Alkaline serine protease Optimization Protein modeling 

Notes

Acknowledgements

This project (Ph.D. Thesis) is financially supported by Tabriz University of Medical Sciences, Tabriz, Iran. Grant Number: 58056.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer Nature B.V. 2020

Authors and Affiliations

  1. 1.Drug Applied Research CenterTabriz University of Medical SciencesTabrizIran
  2. 2.Department of Pharmaceutical Biotechnology, Faculty of PharmacyTabriz University of Medical SciencesTabrizIran
  3. 3.Department of Medical Nanotechnology, Faculty of Advanced Medical ScienceTabriz University of Medical SciencesTabrizIran

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