Purification,structural characterization and biotechnological potential of tannase enzyme produced by Enterobacter cloacae strain 41 |
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| Institution: | 1. Department of Zoology, School of Life Sciences, Periyar University, Salem 636011, Tamil Nadu, India;2. Sri Paramakalyani Centre for Environmental Sciences, Manonmaniam Sundaranar University, Alwarkurichi 627412, Tamil Nadu, India;3. Department of Environmental Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India;4. Functional Genomics Laboratory, Department of Biological Science and Technology, National Pingtung University of Science and Technology, Neipu, Pingtung 91201, Taiwan;5. Department of Biochemistry, Central University of Rajasthan, Ajmer, Rajasthan 305817, India;1. Department of Bio & Nano Technology, Guru Jambheshwar University of Science & Technology, Hisar, India;2. Department of Biotechnology, Maharishi Markandeshwar University, Mullana, India;3. School of Bioengineering and Food Technology, Shoolini University, Solan, Himachal Pradesh, India;4. Department of Biochemistry, Kurukshetra University, Kurukshetra, India;1. Department of Food Science and Technology, Chonbuk National University, Jeonju, 49896, Jeonbuk, Republic of Korea;2. Foundation of Agri. Tech. Commercialization and Transfer, Iksan, 54667, Jeonbuk, Republic of Korea;3. Agricultural Research and Extention Services, Iksan, 54591, Jeonbuk, Republic of Korea;1. Department of Biotechnology, Ambala College of Engineering and Applied Research, Devsthali 133101, Ambala, Haryana, India;2. Department of Biotechnology, Maharishi Markandeshwar University, Mullana 133203, Ambala, Haryana, India;3. Department of Biotechnology, Chaudhary Devi Lal University, Sirsa 125055, Haryana, India;1. Biochemistry Department, Division of Genetic Engineering and Biotechnology, National Research Centre, Dokki 12622, Giza, Egypt;2. Biochemistry Department, Faculty of Science, Ain Shams University, Cairo, Egypt |
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| Abstract: | Tannase production by Enterobacter cloacae strain 41 was investigated under submerged fermentation which was optimized at various circumstances such as pH, temperature, substrate, and agitation, carbon, and nitrogen sources. Tannase was purified by a two-step approach comprising of ion exchange and size exclusion chromatography, respectively. The maximum tannase production was achieved at 1.0% tannic acid concentration, incubation temperature of 50 °C, and initial pH 6.0. The molecular weight of purified tannase was 45 kDa on 10% SDS-PAGE, and it was confirmed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS). The enzymatic products of purified tannase were characterized by HPLC, TLC and FT-IR spectroscopy which showed the functional groups such as OH, CO, and CC. The purified tannase retained the activity up to 90% under the condition at 50 °C and pH 6.0 after 1 h incubation. Enzyme kinetics and inhibition studies were also investigated. Cytotoxicity studies showed that the tannase has no cytotoxic effects on Vero cell line. The results indicated the E. cloacae strain 41 would give a potential source for the efficient production of tannase and can be used in tannery effluent degradation, food, and pharmaceutical industrial applications. |
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| Keywords: | Tannase purification MALDI-TOF-MS FT-IR Circular dichroism |
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