Extraction, purification and biochemical characterization of a peroxidase from Copaifera langsdorffii leaves

Abstract

The aim of this work is to obtain, purify and characterize biochemically a peroxidase from Copaifera langsdorffii leaves (COP). COP was obtained by acetone precipitation followed by ion-exchange chromatography. Purification yielded 3.5% of peroxidase with the purification factor of 46.86. The COP optimum pH is 6.0 and the temperature is 35 ºC. COP was stable in the pH range of 4.5 to 9.3 and at temperatures below 50.0 ºC. The apparent Michaelis-Menten constants (Km) for guaiacol and H2O2 were 0.04 mM and 0.39 mM respectively. Enzyme turnover was 0.075 s-1 for guaiacol and 0.28 s-1 for hydrogen peroxide. Copaifera langsdorffii leaves showed to be a rich source of active peroxidase (COP) during the whole year. COP could replace HRP, the most used peroxidase, in analytical determinations and treatment of industrial effluents at low cost

Catalytic and kinetic properties of purified Eichhornia crassipes leaf peroxidase

“…The specific activity (470 µmol min -1 mg -1 ) of E. crassipes leaf peroxidase achieved after the last step of the three-step purification in this study was considerably higher than levels obtained previously for different sources of the enzyme: 15.21 units mg -1 (Rehman et al 1999), 86 units mg -1 (Regalado et al 1996), 135.44 units mg -1 (Maciel et al 2007), 346.43 units mg -1 (Khatun et al 2012) and 349.8 units mg -1 (Srinivas et al 1999). …”

“…Purified peroxidases have been obtained from diverse plant sources (Deepa, Arumughan 2002), such as spring cabbage, soy bean and rice leaves (Ito et al 1991), tea (Kvaratskhelia et al 1997), okra (Yemenicioglu et al 1998), Ipomoea palmetto (Srinivas et al 1999) and Copaifera longsdorffi (Maciel et al 2007). However, less attention has been directed toward sources that are naturally widely available and regarded as unwanted in the environment.…”

Responses of bioenergy sorghum cell wall metabolism to agronomic practices

“…Peroxidase kinetics and lignin yield: The Km values ( Table 1) of hydrogen peroxide are much lower than earlier reports [25] probably because the inactive isoenzymes of the peroxidase had been removed through isoelectric focusing (Figures 1-4). This further supports the observed increases in the Vmax values.…”

“…Most of the inactive isoenzymes were acidic (pI < 4.0), and the active isoenzymes (boxed) had higher pI values. Chromatographic and two-dimensional purification procedures had been inadequate for concentrating the active isoenzymes, and removing them from the inactive forms [23][24][25]. Native polyacrylamide gel electrophoresis (PAGE) of the Rotofor fractions revealed the two groups (anionic and cationic) of isoenzymes that make up the charge isomers (Figures 1-4).…”

Biochemical characterization of Santalum album (Chandan) leaf peroxidase

“…The supernatant was discarded, and the acetone powder was dried at a temperature of 10-15°C and dissolved in deionized water (Dubey et al 2007;Kumar et al 2008;Singh et al 2010) and it is used for the peroxidase assay and the estimation (Dubey et al 2007). The partial purified peroxidase was stored at −18°C until use (Maciel et al 2007). …”