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Thermoalkalophilic Lipase From an Extremely Halophilic Bacterial Strain Bacillus Atrophaeus Fshm2: Purification, Biochemical Characterization and Application Publisher



Ameri A1 ; Shakibaie M1 ; Faramarzi MA2 ; Ameri A1 ; Amirpourrostami S1 ; Rahimi HR4 ; Forootanfar H5
Authors
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Authors Affiliations
  1. 1. Pharmaceutics Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran
  2. 2. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy and Biotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Department of Medicinal Chemistry, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran
  4. 4. Department of Toxicology and Pharmacology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran
  5. 5. Herbal and Traditional Medicines Research Center, Kerman University of Medical Sciences, Kerman, Iran

Source: Biocatalysis and Biotransformation Published:2017


Abstract

The present study was designed to isolate and identify an extremely halophilic lipase-producing bacterial strain, purify and characterize the related enzyme and evaluate its application for ethyl and methyl valerate synthesis. Among four halophilic isolates, the lipolytic ability of one isolate (identified as Bacillus atrophaeus FSHM2) was confirmed. The enzyme (designated as BaL) was purified using three sequential steps of ethanol precipitation and dialysis, Q-Sepharose XL anion-exchange chromatography and SP Sepharose cation-exchange chromatography with a final yield of 9.9% and a purification factor of 31.8. The purified BaL (Mw∼85 kDa) was most active at 70 °C and pH 9 in the presence of 4 M NaCl and retained 58.7% of its initial activity after 150 min of incubation at 80 °C. The enzyme was inhibited by Cd2+ (35.6 ± 1.7%) but activated by Ca2+ (132.4 ± 2.2%). Evaluation of BaL's stability in the presence of organic solvents showed that xylene (25%) enhanced the relative activity of the enzyme to 334.2 ± 0.6% after 1 h of incubation. The results of esterification studies using the purified BaL revealed that maximum ethyl valerate (88.5%) and methyl valerate (67.5%) synthesis occurred in the organic solvent medium (xylene) after 48 h of incubation at 50 °C. © 2017 Kerman University of Medical Sciences, Kerman, Iran. Published by informa UK Limited, trading as Taylor & Francis Group.
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