Kinetic constants determination for an alkaline protease from Bacillus mojavensis using response surface methodology

Biotechnology and Bioengineering
Q K BegR Gupta

Abstract

The kinetic constants for an alkaline protease from Bacillus mojavensis were determined using a central composite circumscribed design (CCCD) where concentration of substrate (casein) and the assay temperature were varied around their center point. The K(m),V(max), K(cat), activation energy (E(a)) and temperature coefficient (q(10)) were determined and the values of these kinetic constants obtained were found comparable to that obtained with conventional methods. The Michaelis-Menten constant (K(m)) for casein decreased with corresponding increase in V(max), as reaction temperature was raised from 45-60 degrees C. The protease exhibited K(m) of 0.0357 mg/ml, 0.0270 mg/ml, 0.0259 mg/ml, and 0.0250 mg/ml at 45, 50, 55, and 60 degrees C, respectively, whereas V(max) values at these temperatures were 74.07, 99.01, 116.28, and 120.48 microg/ml/min, respectively, as determined by response surface methodology. The Arrhenius plot suggested that the enzyme undergoes thermal activation above 45 degrees C until 60-65 degrees C followed by thermal inactivation. Likewise, the energy of activation (E(a)) was more between 45-55 degrees C (9747 cal/mol) compared to E(a) between 50-60 degrees C (4162 cal/mol).

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Citations

May 9, 2008·Journal of Industrial Microbiology & Biotechnology·Bo-Yuan Chen, Han-Tsung Wang
Apr 9, 2013·Applied Biochemistry and Biotechnology·Hamid Reza Karbalaei-HeidariGhodratollah Absalan
Aug 11, 2012·Bioscience, Biotechnology, and Biochemistry·Mohammad Norazmi AhmadMamot Said
May 31, 2012·Critical Reviews in Biotechnology·Rani GuptaQasim K Beg
Jan 22, 2014·Food Chemistry·Yongguang GuanMingshun Chen
Jan 19, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Fang DongZiyin Yang
Jan 15, 2015·Journal of the Science of Food and Agriculture·Raúl Pérez GálvezAntonio Guadix
Jul 16, 2020·Journal of Environmental Science and Health. Part A, Toxic/hazardous Substances & Environmental Engineering·Tanvir Hossain EmonAbul Kalam Azad
Aug 20, 2018·Environmental Science and Pollution Research International·Senbagam DuraisamyAnbarasu Kumarasamy
Jul 3, 2019·Frontiers in Bioengineering and Biotechnology·Abdul RazzaqMuhammad Ashraf
Sep 22, 2020·3 Biotech·Juliana Mota de OliveiraSandra Aparecida de Assis

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