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Biochemical characterization of recombinant benzyl alcohol dehydrogenase from Rhodococcus ruber UKMP-5M
2017
Tavakoli, Arezoo | Hamzah, Ainon
Benzyl Alcohol Dehydrogenase (BADH) is an important enzyme for hydrocarbon degradation, which can oxidize benzyl alcohols to aldehydes, while being capable of catalyzing a reversible reaction by reducing benzaldehyde. BADH is a member of medium chain alcohol dehydrogenases, in which zinc and NAD are essential for enzyme activity. This paper describes the expression, purification, and characterization of recombinant benzyl alcohol dehydrogenase, encoded by xylB gene from Rhodococcus ruber UKMP-5M. The gene has been amplified and cloned into E. coli, and the recombinant plasmid pGEMT-xylB has been digested by NdeI and HindIII to construct plasmid pET28b-xylC and then ligated into E. coli BL21 (DE3), itself induced by 0.3 mM isopropyl β-D-thiogalactoside (IPTG) at 25°C. The expressed BADH has been 38 kDa, and is purified by affinity chromatography, in which the specific activity was 30 U/mg after 17 folds purification, leading to a NAD-dependent enzyme that uses benzyl alcohol as a substrate for enzyme characterization. The final metabolite is benzaldehyde, identified by gas chromatography mass spectrometry (GC-MS). The BADH activity has been 0.7 U/mL and the optimum pH and temperature, 9.5 and 30ºC, respectively. Also the Michaelis constant (Km) and maximum velocity (Vmax) have accounted to 705 µM and 1.3 U/mL, respectively. Benzyl alcohol dehydrogenase from R. ruber UKMP-5M can be used for hydrocarbon biodegradation in contaminated sites.
Afficher plus [+] Moins [-]Overhauling health effects perspectives.
1988
Schwebach G.H. | Cafaro D. | Egan J. | Grimes M. | Michael G.
Water pollution | State revolving funds insufficient to meet wastewater treatment needs | State revolving fund program
1992
Water pollution | Nonindustrial wastewater pollution can be better managed
1991
Decontamination and energetic supply in rural installations
1992
Carbonell, V. (Escuela Tecnica Superior de Ingenieros Agronomos, Madrid (Spain). Dept. de Fisica) | Bigeriego, M. | Delgado, M.
Common treatment of sewage sludge and animal waste slurries in covered digesters
1992
Linke, B. | Kolisch, G. (Essen Univ. (Germany). Inst. fuer Umwelttechnik)
Effects of the recirculation on the anaerobic digestion of cow manure in a biofilm reactor
1992
Sanchez Hernandez, E.P. (National Center for Scientific Research, La Habana (Cuba). Environmental Pollution Dept.) | Weiland, P. | Travieso Cordoba, L.
Purification and characterization of biosurfactant produced by Bacillus licheniformis Y-1 and its application in remediation of petroleum contaminated soil
2016
Liu, Boqun | Liu, Jinpeng | Ju, Meiting | Li, Xiaojing | Yu, Qilin
In our previous research, a petroleum degrading bacteria strain Bacillus licheniformis Y-1 was obtained in Dagang Oilfield which had the capability of producing biosurfactant. This biosurfactant was isolated and purified in this work. The biosurfactant produced by strain Y-1 had the capability to decrease the surface tension of water from 74.66 to 27.26mN/m, with the critical micelle concentration (CMC) of 40mg/L. The biosurfactant performed not only excellent stabilities against pH, temperature and salinity, but also great emulsifying activities to different kinds of oil, especially the crude oil. According to the results of FT-IR spectrum and 1H NMR spectrum detection, the surfactant was determined to be a cyclic lipopeptide. Furthermore, through the addition of surfactant, the effect of petroleum contaminated soil remediation by fungi got a significant improvement.
Afficher plus [+] Moins [-]Small water system treatment costs
1986
Gumerman, Robert C. | Burris, Bruce E. | Hansen, Sigurd P.
Nitrogen oxides emission control, 1972
1972
Lawrence, A. A.
