Purification and identification of cutinases from Colletotrichum kahawae and Colletotrichum gloeosporioides

文献类型： 外文期刊

第一作者： Chen, Zhenjia

作者： Chen, Zhenjia;Franco, Catarina F.;Baptista, Ricardo P.;Cabral, Joaquim M. S.;Coelho, Ana V.;Rodrigues, Carlos J., Jr.;Melo, Eduardo P.

作者机构：

关键词： Colletotrichum kahawae;Colletotrichum gloeosporioides;carboxylesterase;cutinase;protein purification;mass spectrometry;DESORPTION/IONIZATION MASS-SPECTROMETRY;PROTEIN;SEQUENCE;ENZYME;ACIDS;GENE

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Colletotrichum kahawae is the causal agent of the coffee berry disease, infecting leaves and coffee berries at any stage of their development. Colletotrichum gloeosporioides is the causal agent of brown blight, infecting ripe berries only. Both fungi secrete the same pattern of carboxylesterases to the fermentation broth when cutin is used as carbon source. By using two different strategies composed of two precipitation steps (ammonium sulphate and acetic acid precipitation) and two chromatographic steps, two proteins displaying carboxylesterase activity were purified to electrophoretic homogeneity. One, with a molecular weight (MW) of 21 kDa, has a blocked N terminus and was identified as cutinase by peptide mass fingerprint and mass spectrometry/mass spectrometry data acquired after peptide derivatization with 4-sulphophenyl isothiocyanate. The second, with a MW of 40 kDa, displays significant carboxylesterase activity on tributyrin but low activity on p-nitrophenyl butyrate. N-terminal sequencing for this protein does not reveal any homology to other carboxylesterases. These two enzymes, which were secreted by both fungi, appear homologous.

分类号： Q93

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