Crystallization and preliminary X-ray diffraction analysis of a putative carbon-carbon bond hydrolase from Mycobacterium abscessus 103

摘要

The PhlG protein from Mycobacterium abscessus 103 (mPhlG), which shares 30% sequence identity with phloretin hydrolase from Eubacterium ramulus and 38% sequence identity with 2,4-diacetylphloroglucinol hydrolase from Pseudomonas fluorescens Pf-5, is a putative carbon-carbon bond hydrolase. Here, the expression, purification and crystallization of mPhlG are reported. Crystals were obtained using a precipitant consisting of 100 mM citric acid pH 5.0, 1.0 M lithium chloride, 8%(w/v) polyethylene glycol 6000. The crystals diffracted to 1.87 angstrom resolution and belonged to space group P2(1), with unit-cell parameters a = 71.0, b = 63.4, c = 74.7 angstrom, alpha = 90.0, beta = 103.2, gamma = 90.0 degrees. Assuming the presence of two mPhlG molecules in the asymmetric unit, V-M was calculated to be 2.5 angstrom(3) Da(-1), which corresponds to a solvent content of 50%.