Open AccessExpression and homology modeling of sterol 14α‐demethylase from Penicillium digitatum
Author(s) -
Zhao Li,
Liu Deli,
Zhang Qingye,
Zhang Shan,
Wan Jian,
Xiao Wenjing
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2007.00929.x
Subject(s) - penicillium digitatum , biochemistry , biology , escherichia coli , microbiology and biotechnology , chemistry , postharvest , botany , gene
Green mold of citrus, caused by Penicillium digitatum , is the most serious postharvest disease of citrus. Sterol 14α‐demethylase (CYP51) is one of the key enzymes of sterol biosynthesis in biological kingdoms and is a prime target of antifungal drugs. To exploit novel 14α‐demethylase inhibitor (DMI) fungicides, DNA and total RNA were isolated from P. digitatum . The CYP51 of P. digitatum was cloned and expressed in Escherichia coli , yielding recombinant protein with a molecular weight of c . 59 kDa. The P. digitatum CYP51 protein (PdCYP51) was purified and polyclonal antibodies were prepared. Compared with the sequence of P. digitatum PD5 in GenBank, there were four mutated nucleotides which resulted in four mutated amino acids. The three‐dimensional (3D) model of P. digitatum CYP51 was established based on structure template of 1e9x.pdb and diniconazole was docked into the active site by FlexX. According to spectral data, it is suggested that the purified soluble protein had high affinity with diniconazole, a potent inhibitor of CYP51 reaction in fungi. At the same time, these spectral data suggested that the 3D model and the docking model were reasonable, which we hope can be used to provide a virtual screening of novel DMI drugs.