Akademik Veri Yönetim Sistemi
V. International Enzyme and Bioprocess Days EBDays 2024 , İzmir, Türkiye, 27 - 29 Ağustos 2024, ss.97
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.