Polyextremotolerant Amylase Produced from Novel Enterococcus with Potpourri of Applications
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Amylolytic toluene-tolerant Enterococcus faecalis mercadA7 isolated from asbestos contaminated black soil from the Tokat province of Turkey was used for statistically optimized liquid phase fermentation to spawn elevated levels of amylase. Box Behnken technique of Response surface Methodology is followed to optimize three significant factors influencing Amylase production. To purify the amylase, ammonium sulfate fractionation, dialysis and gel filtration on Sephadex G-100 column chromatography were used. Submerged state fermentation at statistically optimized conditions augmented amylase production levels (8467 U ml−1) 8.74-fold compared to the conventional optimization. Purified amylase was found to possess specific activity of 1022 U mg−1, optimal activity at pH 9 and 60 °C, 80% stable at 8.0–10.0 pH range and 90% stable at 45–75 °C for 1 h; with Km value of 8.06 mg ml−1 and a Vmax of 4.36 mmol h−1 ml−1, noteworthy stability up to 43% toluene, and upto 8% NaCl. High titers of Amylase produced from Enterococcus faecalis mercadA7 might find applications in copious industrial processes due to noteworthy stability of the cell-free purified enzyme at elevated temperatures, alkaline, and high toluene or salt concentrated environments.
KeywordsAmylase Toluene Enterococcus faecalis mercadA7 Purification Characterization
Financial assistance by TUBITAK, The Scientific and Technological Research Council of Turkey Grants Scholarships for 2216—Research Fellowship Program for International Researchers program (2015–2016), Grant Number: 21514107-115.99-129944 is deeply acknowledged by recipient Dr. Haritha Meruvu (Postdoctoral Research Scientist).
Compliance with Ethical Standards
Conflict of interest
The author declares no conflict of interest.
Research Involving Human Participants and/or Animals
No humans or animals are used or involved; no ethical issues, this work is done by the sole and corresponding author.
- Box GEP, Hunter WG, Hunter JS (1978) Statistics for experiments. Wiley, New York, pp 291–334Google Scholar
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:267–275Google Scholar
- Mahmoud MH (2018) Production, purification and characterization of alfa-amylase produced by bacteria from Pharaonic Lake. J Bas Environ Sci 5:162–173Google Scholar
- Pandey S, Sharma AK, Solanki KP, Singh SP (2018) Catalysis and stability of an extracellular α-amylase from a haloalkaliphilic bacterium as a function of the organic solvents at different pH, salt concentrations and temperaturas. IJMS 47(01):240–248Google Scholar
- Sethi BK, Nanda PK, Sahoo S, Sena S (2016) Characterization of purified α-amylase produced by Aspergillus terreus NCFT 4269.10 using pearl millet as substrate. Cogent Food Agric 2(1):1158902Google Scholar