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Catalytic Profile and Amylolytic Studies of Toluene-Tolerant Enterococcus faecalis str. nov. mercadA7

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Abstract

This work pivots around screening a toluene-tolerant bacterium with concomitant \(\upalpha \)-amylase production ability. Eighteen \(\upalpha \)-amylase-producing bacterial strains were isolated from samples collected from Tokat province of Turkey among which strain A7 showed maximum \(\upalpha \)-amylase production titers (968 U/ml). The strain A7 was identified as Enterococcus faecalis mercadA7 (KX298857) with 54% GC content and 98% similarity to the closest strain, Enterococcus faecalis NBRC100480 through 16srRNA typing studies. The amylase production was improved 2.6-folds compared to the basal medium at optimal conditions recorded: \(37\,{^{\circ }}\)C, pH 7.0, 18 h incubation, 1%v/v inoculum content and 20% toluene (carbon) supplementation. The amylase activity was demonstrated to increase upon exposure to toluene by a yet to be determined mechanism. The various growth kinetic parameters recorded are specific growth rate, \(\mu _{\mathrm{max}}\) of 0.318 \(\hbox {h}^{-1}\); constant, \(K_{{s}}\) 10.48 mg/ml; yield coefficient, \(Y_{{x}/{s}}\) of 0.1157 mg/g; and cell doubling time, \(t_{\mathrm{d}}\) of 51 min. The enzyme was partially purified with a threefold increase in specific activity; it was found to be active at pH 8 and \(40\,{^{\circ }}\)C; stable from 30 to \(50\,{^{\circ }}\)C and 7–10 pH range; maintaining stability up to 40% of toluene. The purified enzyme’s maximum velocity, \(V_{\mathrm{max}}\) of 3.33 \(\hbox {mmol}/\hbox {h}\,\hbox {ml}\) with constant, Km of 5 mg/ml were determined by Line Weaver Burke plot. Both Enterococcus faecalis mercadA7 and the purified \(\upalpha \)-amylase might find applications in several industrial processes due to the catalytic promiscuity triggered by the presence of toluene, and augmented stability of the cell-free purified enzyme as well as the organism to high alkaline/toluene levels.

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  1. Department of BioEngineering, Faculty of Engineering and Natural Sciences, Gaziosmanpasha University, Tokat, Turkey

    Haritha Meruvu

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Meruvu, H. Catalytic Profile and Amylolytic Studies of Toluene-Tolerant Enterococcus faecalis str. nov. mercadA7. Arab J Sci Eng 42, 1517–1527 (2017). https://doi.org/10.1007/s13369-016-2386-x

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