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Two laccase temperature isoforms capable of oxidizing phenolic compounds to quinones were isolated and purified to homogeneity from the cladodes of the xerophyte species Opuntia vulgaris. These catalytically active proteins exhibit apparent molecular masses of 137 and 90 kDa. Under reducing conditions, both isoforms yielded a subunit molecular mass of 43 kDa, suggesting that the enzyme is a multimer of the 43 kDa subunit. The 137 kDa isoform when heated at 80 degrees C for 3min generated three polypeptide bands on activity stained polyacrylamide gels exhibiting 137, 90 and 43 kDa molecular forms. All isoforms of the enzyme exhibited an optimum pH of 10 when 2,6-dimethoxyphenol was used as a substrate. The optimum temperature of the 137 kDa enzyme form was noted to be 80 degrees C and that of the 90 kDa enzyme form was 70 degrees C. Denaturation kinetics of both the laccase isoforms carried out at their respective optimum temperatures for 30 min exhibited enzyme activity in excess of their t(1/2) values throughout the assay period. The K(m) for the 137 kDa form was determined to be 2.2 +/- 0.3 mm and the V(max) was 2.8 +/- 0.2 IU/mL. These high temperature stable laccase isoforms having alkaline pH optima can find significant industrial use. Copyright (c) 2010 John Wiley & Sons, Ltd.
Department of Biochemistry and Molecular Biology, School of Life Sciences, Pondicherry University, Puducherry 605014, India.
This article was published in the following journal.
Name: Biomedical chromatography : BMC
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A plant genus of the family CACTACEAE. Species with cylindrical joints are called Cholla; flat jointed ones are Prickly-pear.
A species of gram-negative bacteria and nitrogen innoculant of PHASEOLUS VULGARIS.
A species of green microalgae in the family Chlorellaceae. It is used as a model organism for PHOTOSYNTHESIS, and as a food supplement (DIETARY SUPPLEMENTS).
A species of the Beta genus. Cultivars are used as a source of beets (root) or chard (leaves).
A species of gram-negative, anaerobic, rod-shaped bacteria isolated from soil, animal intestines and feces, and fresh and salt water.
Enzymes are proteins that catalyze (i.e., increase the rates of) chemical reactions. In enzymatic reactions, the molecules at the beginning of the process, called substrates, are converted into different molecules, called products. Almost all chemical re...
An assay is an analytic procedure for qualitatively assessing or quantitatively measuring the presence or amount or the functional activity of a target entity. This can be a drug or biochemical substance or a cell in an organism or organic sample. ...