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Statistical optimization and production of protease from estuarine Citrobacter diversus

Author Affiliations

  • 1Faculty of Marine Sciences, Annamalai University, Parangipettai, Tamil Nadu, India
  • 2Faculty of Marine Sciences, Annamalai University, Parangipettai, Tamil Nadu, India
  • 3Indian Institute of Chemical Technology, Mumbai, India
  • 4Indian Institute of Chemical Technology, Mumbai, India

Int. Res. J. Biological Sci., Volume 5, Issue (5), Pages 18-25, May,10 (2016)


The present study is on protease enzyme production by a Citrobacter diversus strain which was isolated from sediment sample of Vellar Estuary, Tamil Nadu, India. Response surface methodology (RSM) was employed to optimize the production medium for increasing protease production. Plackett–Burman experimental design was used in the first step of screening the media component of optimization, Coconut oil cake (Cheaper carbon source), yeast extract, temperature, pH, salinity and incubation period were found to be significant factors affecting protease production. To determine the optimal concentration of each significant variable, a central composite design was employed. Based on response surface and analysis of variance, the optimum concentrations of the components were obtained as follows: coconut oil cake (3.24 %), yeast extract (0.669 %), temperature (35.20C), pH (7.46), salinity (16.40 ppt) and incubation period (50.88 hrs). The maximum protease activity of 92.93 U/ml/min was predicted by the model. The Citrobacter diversus produced 92.4 U/ml/min of protease.


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