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Scale-up of protease production by Serratia marcescens using municipal solid wastes in the bioreactor and its partial purification and characterization

Mohammad Rejaur Rahman, Md. Saddam Hossain, Akhikun Nahar,Md. Faisal Azim, Kamrul Islam, Abul Kalam Azad

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Int. J. Biosci.12(4), 99-109, April 2018

DOI: http://dx.doi.org/10.12692/ijb/20.2.1-20

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A proteolytic bacterial isolate obtained from municipal solid wastes (MSW), was identified as Serratia marcescens based on cultural, morphological and biochemical characteristics. In the present study, protease production by S. marcescens was maximized using MSW as the sole source of carbon and nitrogen under some optimized physicochemical conditions. The scale-up of protease production with different concentrations of organic MSW was performed. In shake flask fermentation, maximum level of protease was produced with 4 % MSW after 48 h at 30 °C, 120 rpm and pH 8.0. However, in the bioreactor, optimum level of protease was produced with 3 % MSW after 24 h of fermentation at 120 rpm, 30 °C and pH 8.0. In comparison with the shake flask, protease production was scaled-up 2 fold in the bioreactor with reduction in fermentation period. Partial purification by ammonium sulfate fractionation and anion-exchange chromatography resulted in a final 37-fold purified protease with a specific activity of 9411 U/mg protein and a typical yield of 7.1 %. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the estimated molecular mass of the partially purified protease was around 55 kDa. The partially purified protease showed optimum activity at pH 7.0 and 40 °C.  The purified protease was stable at pH 5.0 – 9.0 and temperatures up to 45 °C. The enzyme activity was stimulated by Mg2+, K+ and Ca2+ but was severely inhibited by Zn2+ and Hg2+.


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Scale-up of protease production by Serratia marcescens using municipal solid wastes in the bioreactor and its partial purification and characterization

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