Abstract
Protease has been used in large application industrial process such as detergent, leather, textil, softdrink, andmedicine. In order to find unique protease, many substances were explored as proteases of bacteria sources. Inthis study, tofu liquid waste was used as a source of bacteria producing proteases. Waste sample was growth inskim milk agar medium showing proteases activity, it was used to produce extracellular protease. The microbialcolonies were identified as Staphyllococcus sp. Protease was extracted with 5000 g centrifugation at 4 0C, andpurificated with ammonium sulphate precipitation continued with dialisis. Optimum production time, pH, metal ion,EDTA, specific activity, KM, and Vmaks were studied for enzyme characterization. Volume of crude enzyme was 300ml, with spesific activity of 3.55 U/mg. Protease obtained from 60% ammonium sulphate fraction had the highestspecific activity of 68.22 U/mg. Study on the protease characterization revealed that optimum temperature of thisenzyme was 400C. The optimum pH of the enzyme was found to be 8.0. The kinetic parameters K M dan Vmaks withcasein as substrate were 0.31% and 51.55 U/ml. Some inhibitory effect was observed in the presence of EDTA, Cu +2,Co+2, Zn+2, and enzyme activity was stimulated by Mg+2, indicating that this ion had a functional role in the molecularstructure of the enzyme.
Highlights
EDTA, specific activity, K, and V were studied for enzyme characterization
Protease obtained from 60% ammonium sulphate fraction had the highest specific activity of 68.22 U/mg
Some inhibitory effect was observed in the presence of EDTA, Cu+2, Co+2, Zn+2, and enzyme activity was stimulated by Mg+2, indicating that this ion had a functional role in the molecular structure of the enzyme
Summary
Sebanyak 0,1 mL larutan enzim ditambahkan ke dalam substrat dan diinkubasi pada suhu 350C selama 30 menit. Aktivitas protease (U) didefinisikan sebagai jumlah enzim yang diperlukan untuk menghasilkan 1 μg tirosina (ekuivalen)/menit/ml larutan enzim dari substrat kasein pada kondisi pengujian tersebut. Prosedur kerja penetapan pH optimum enzim sama dengan seperti pada uji aktivitas protease, tetapi dengan variasi pH. Prosedur kerja penentuan suhu optimum enzim sama seperti uji aktivitas protease dengan variasi suhu 30; 35; 40; 45, 50; 55; 60; dan 650C, inkubasi dilakukan bergantian. Penentuan Vmaks dan KM dilakukan dengan menguji aktivitas protease pada kondisi pH dan suhu optimum dengan variasi konsentrasi substrat kasein yaitu 0,5; 1; 1,5; 2,0 dan 2,5%. M sebanyak 0,06 ml ke dalam larutan sampel sehingga konsentrasi akhir larutan pada saat uji aktivitas dilakukan adalah 10-3 M.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
