Predicted secondary structure of hydroperoxide lyase from green bell pepper cloned in the yeast Yarrowia lipolytica

被引:8
作者
Santiago-Gomez, Mirna P. [1 ]
Kermasha, Selim [2 ]
Nicaud, Jean-Marc [3 ]
Belin, Jean-Marc [1 ]
Husson, Florence [1 ]
机构
[1] Univ Bourgogne, Lab GPMA, F-21000 Dijon, France
[2] McGill Univ, Dept Food Sci & Agr Chem, Ste Anne De Bellevue, PQ H9X 3V9, Canada
[3] INRA, CNRS, INAPG, Lab Microbiol & Genet Mol, F-78850 Thiverval Grignon, France
关键词
Hydroperoxide lyase; Yarrowia lipolytica; Secondary structure; Prediction; Circular dichroism spectroscopy; 13-HYDROPEROXIDE LYASE; AMINO-ACID; PURIFICATION; CONFORMATION; PROTEINS; CLONING; EXPRESSION; SEQUENCES; FRUITS;
D O I
10.1016/j.molcatb.2010.01.009
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Fatty acid hydroperoxide lyase (HPL) is a member of the cytochrome P450 family acting on fatty acid hydroperoxides in many organisms. The active green bell pepper HPL, cloned and expressed in the yeast Yarrowia lipolytica, was purified by immobilized metal-ion affinity chromatography (IMAC) in the presence of 2% of Triton X-100R. The secondary structure prediction by bioinformatics servers of HPL was realized by ANTHEPROT software, using the GOR, DPM and Predator methods. The theoretical results which are average values obtained from three different calculation methods showed 33% alpha-helix, 18% beta-sheet, 7% turn and 42% coil. On the other hand, the secondary structure approach of the purified active HPL (specific activity of 2.94 U/mg protein) was realized by differential scanning calorimetry (DSC) and circular dichroism (CD) spectroscopy, and showed 13% alpha-helix, 29% beta-sheet, 5% turn and 53% random coil. (C) 2010 Elsevier B.V. All rights reserved.
引用
收藏
页码:63 / 67
页数:5
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