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Screening and Selection of Marine Isolate for l-Glutaminase Production and Media Optimization Using Response Surface Methodology

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Abstract

The current work details the screening of about 400 marine isolates from various marine niches, from which one isolate was finally selected based on the productivity of glutaminase (71.23 U/l). Further, biochemical identification tests and 16S rRNA sequencing identified this isolate to be Providencia sp. This isolate was taken up for further media optimization studies by using one-factor-at-a-time approach and subsequently by response surface methodology. A face centered central composite design was employed to investigate the interactive effects of four variables, viz., concentrations of glucose, methionine, urea, and succinic acid on glutaminase production. A significant influence of urea on glutaminase production was noted. Response surface methodology showed that a medium containing (g/l) glucose 10.0, urea 5.15, methionine 3.5, succinic acid 6.0, ammonium sulfate 2.5, and yeast extract 6.0 to be optimum for the production of glutaminase. The applied methodology was validated using this optimized media and enzyme activity 119 ± 0.12 U/l and specific activity of 0.63 U/mg protein after 28 h of incubation at 25 °C was obtained.

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Acknowledgment

The authors gratefully acknowledge Dr. Yogesh Shouche and his team at NCCS, Pune, India for 16S rRNA sequencing of the isolate and to Department of Biotechnology (DBT), Government of India for financially supporting this work.

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  1. Food Engineering and Technology Department, Institute of Chemical Technology, Nathalal Parikh Marg, Matunga, Mumbai, 400019, India

    Padma V. Iyer & Rekha S. Singhal

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  1. Padma V. Iyer
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  2. Rekha S. Singhal
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Correspondence to Padma V. Iyer.

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Iyer, P.V., Singhal, R.S. Screening and Selection of Marine Isolate for l-Glutaminase Production and Media Optimization Using Response Surface Methodology. Appl Biochem Biotechnol 159, 233–250 (2009). https://doi.org/10.1007/s12010-008-8522-7

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  • DOI: https://doi.org/10.1007/s12010-008-8522-7

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