Cloning and Enhanced Expression of an Extracellular Alkaline Protease from a Soil Isolate of Bacillus clausii in Bacillus subtilis
نویسندگان: ثبت نشده
چکیده مقاله:
in the detergent industry. In this study, the extracellular alkaline serine protease gene, aprE, from Bacillusclausii was amplified by PCR and further cloned and expressed in B. subtilis WB600 using the pWB980 expression vector. Protease activity of the recombinant B. subtilis WB600 harboring the plasmid pWB980/aprEreached up to 1020 U/ml, approximately 3-folds higher than the native B. clausii strain. Characterization of the recombinant alkaline protease by SDS-PAGE and zymogram analyses indicated a molecular weight of31 kDa. DNA sequence analysis and the deduced amino acid sequence revealed 98% homology with theextracellular alkaline serine protease from B. clausii KSM-K16.
منابع مشابه
cloning of alkaline protease gene from bacillus subtilis 168
the aim of this study was to clone the serine alkaline protease-encoding gene from bacillus subtilis 168. this protease, which can have many applications especially in detergent, may be industrially an important enzyme. for the amplification of the gene, pcr was performed with a pair of primers specifically designed for this purpose. electrophoresis of the pcr product showed the expected band o...
متن کاملIsolation, Purification and Characterization of a Thermophilic Alkaline Protease from Bacillus subtilis BP-36
The goal of this research was to isolate and identify the thermostable alkaline protease producing bacteria among several native Iranian microorganisms. At the end of screening program, a Bacillus subtilis BP-36 strain producing thermophilic alkaline protease was isolated from a hot spring in Ardebil province. The target enzyme was purified using a one-step Aqueous two-phase systems (ATPS) prot...
متن کاملMOLECULAR CLONING AND EVALUATION OF WILD PROMOTER IN EXPRESSION OF BACILLUS SPHAERICUS PHENYLALANINE DEHYDROGENASE GENE IN BACILLUS SUBTILIS CELLS
To evaluate the role of wild promoter of L-phenylalanine dehydrogenase (PheDH) gene, referred to as pdh, from Bacillus sphaericus in expression, cloning of pdh gene in Bacillus subtilis was performed. The whole pdh gene was cloned in pHY300PLK shuttle vector and amplified, construct (pHYDH) then transformed in B. subtilis ISW1214 and E. coli JM109. The pdh endogenous promoter presented no effec...
متن کاملProduction and optimization of extracellular Alkaline Protease from Bacillus subtilis isolated from dairy effluent
Proteolytic bacteria are most important for industries such as food and fermentation. Thus isolating and manipulating pure culture from various sources has importance for various biotechnology industries. In the present investigation a total of 26 bacteria were isolated from the dairy plant effluent. All isolates were screened for proteolytic activity using alkaline skim milk agar plate, among ...
متن کاملA study of extracellular alkaline protease from Bacillus subtilis NCIM 2713.
An alkaline protease was isolated from culture filtrate of B. subtilis NCIM 2713 by ammonium sulphate precipitation and was purified by gel filtration. With casein as a substrate, the proteolytic activity of the purified protease was found to be optimal at pH 8.0 and temperature 70 degrees C. The purified protease had molecular weight 20 kDa, Isoelectric point 5.2 and km 2.5 mg ml(-1). The enzy...
متن کاملمنابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ذخیره در منابع من قبلا به منابع من ذحیره شده{@ msg_add @}
عنوان ژورنال
دوره 9 شماره 4
صفحات 275- 280
تاریخ انتشار 2011-10-01
با دنبال کردن یک ژورنال هنگامی که شماره جدید این ژورنال منتشر می شود به شما از طریق ایمیل اطلاع داده می شود.
میزبانی شده توسط پلتفرم ابری doprax.com
copyright © 2015-2023