Isolation and determination of a novel protease for detergent industry from Bacillus velezensis C37plca


Çanakçı S., Ceylan E., Çolak D. N., Beldüz A. O., Nalçaoğlu Şenocak A., Güler H. İ., ...More

V. International Enzyme and Bioprocess Days EBDays 2024 , İzmir, Turkey, 27 - 29 August 2024, pp.97

  • Publication Type: Conference Paper / Summary Text
  • City: İzmir
  • Country: Turkey
  • Page Numbers: pp.97
  • Karadeniz Technical University Affiliated: Yes

Abstract

Proteases are one of the most important industrial enzymes produced by wide range of microorganisms such as bacteria, yeasts and are also found in plants and various animal tissues. Bacterial proteases are mostly extracellular, easily produced in larger amounts, thermostable, active at wider pH range. These properties make the bacterial proteases most suitable for wider industrial application. Alkaline proteases from the bacterial origin are the most important industrial enzymes, which contribute about 60% of the total world enzyme market. The aim of this study is to isolate and characterize bacterial strains from soil and to explore novel source of protease for application in detergent industry. Protease producing several bacteria were isolated from soil contaminated with household waste. As a result of genome analysis (Bacillus velezensis C37plca), 30 different protease genes were identified. To select the protease gene with the desired properties (alkaline, detergent compatible), MALDİ-TOF analysis was performed on the active fractions after ammonium sulfate precipitation and ion chromatography. The gene sequence (fibrinolytic protease) obtained as a result of the analysis was amplified by PCR and cloned into the expression vector, but no activity was observed. Because the accumulation of alkaline protease in the periplasmic space was toxic to E. coli, this might likely be the result of the recombinant strains' incapacity to over-express the enzyme due to lysis. Studies in which alternative methods (changing the host cell, temperatures or expression vector) will be used to investigate the structure-function relationships of the enzyme and solve the activity problem are ongoing.