Partial purification and characterization of serine protease produced through fermentation of organic municipal solid wastes by Serratia marcescens A3 and Pseudomonas putida A2

Publication date: June 2018Source: Journal of Genetic Engineering and Biotechnology, Volume 16, Issue 1Author(s): Asif Iqbal, Al Hakim, Md. Saddam Hossain, Mohammad Rejaur Rahman, Kamrul Islam, Md. Faisal Azim, Jahed Ahmed, Md. Assaduzzaman, Md. Mozammel Hoq, Abul Kalam AzadAbstractProteolytic bacteria isolated from municipal solid wastes (MSW) were identified as Serratia marcescens A3 and Pseudomonas putida A2 based on 16S rDNA sequencing. Protease produced through fermentation of organic MSW by these bacteria under some optimized physicochemical parameters was partially purified and characterized. The estimated molecular mass of the partially purified protease from S. marcescens and P. putida was approximately 25 and 38 kDa, respectively. Protease from both sources showed low Km 0.3 and 0.5 mg ml−1 and high Vmax 333 and 500 µmole min−1 at 40 °C, and thermodynamics analysis suggested formation of ordered enzyme-substrate (E-S) complexes. The activation energy (Ea) and temperature quotient (Q10) of protease from S. marcescens and P. putida were 16.2 and 19.9 kJ/mol, and 1.4 and 1.3 at temperature range from 20 to 40 °C, respectively. Protease of the both bacterial isolates was serine and cysteine type. The protease retained approximately 97% of activity in the presence of sodium dodecyl sulphate. It was observed that the purified protease of S. marcescens could remove blood stains from white cotton cloth and degrade chicken flesh remarkably. Our study revealed that o...
Source: Journal of Genetic Engineering and Biotechnology - Category: Biotechnology Source Type: research