Substrate promiscuity of RdCCD1, a carotenoid cleavage oxygenase from Rosa damascena
Substrate promiscuity of RdCCD1, a carotenoid cleavage oxygenase from Rosa damascena
复制标题
DOI：
10.1016/j.phytochem.2009.01.020
复制
发表时间：
2009-03-01
期刊：
影响因子：
3.8
通讯作者：
Schwab, Wilfried
中科院分区：
文献类型：
--
作者：
Huang, Fong-Chin;Horvath, Gyoergyi;Schwab, Wilfried
玫瑰精油中的几种关键风味化合物是C-13-去甲类异戊二烯，如β-大马酮、β-大马酮和β-紫罗兰酮，它们来源于类胡萝卜素降解。为了寻找类胡萝卜素裂解相关基因，采用简并引物法克隆了蔷薇属植物类胡萝卜素裂解（双）加氧酶（CCD）基因，获得了其全长cDNA（RdCCD 1）。RdCCD 1基因在大肠杆菌中表达，并测定重组蛋白与多种类胡萝卜素底物的切割活性。RdCCD 1蛋白能够在9-10和9 '-10'位置切割多种类胡萝卜素，产生C-14二醛和两种C-13产物，其根据类胡萝卜素底物而变化。RdCCD 1还可以在5-6和5 '-6'位置切割番茄红素，生成6-甲基-5-庚烯-2-酮。利用实时荧光定量PCR技术研究了RdCCD 1基因在月季不同组织中的表达。RdCCDI转录本主要存在于玫瑰花中，其中产生高水平的挥发性C-13-降异戊二烯。因此，玫瑰花中C-13-降异戊二烯类的积累与RdCCD 1的表达相关。(C)2009爱思唯尔有限公司保留所有权利。
Several of the key flavor compounds in rose essential oil are C-13-norisoprenoids, such as P-damascenone, beta-damascone, and beta-ionone which are derived from carotenoid degradation. To search for genes putatively responsible for the cleavage of carotenoids, cloning of carotenoid cleavage (di-)oxygenase (CCD) genes from Rosa damascena was carried out by a degenerate primer approach and yielded a full-length cDNA (RdCCD1). The RdCCD1 gene was expressed in Escherichia coli and recombinant protein was assayed for its cleavage activity with a multitude of carotenoid substrates. The RdCCD1 protein was able to cleave a variety of carotenoids at the 9-10 and 9'-10' positions to produce a C-14 dialdehyde and two C-13 products, which vary depending on the carotenoid substrates. RdCCD1 could also cleave lycopene at the 5-6 and 5'-6' positions to produce 6-methyl-5-hepten-2-one. Expression of RdCCD1 was studied by real-time PCR in different tissues of rose. The RdCCDI transcript was present predominantly in rose flower, where high levels of volatile C-13-norisoprenoids are produced. Thus, the accumulation of C-13-norisoprenoids in rose flower is correlated to the expression of RdCCD1. (C) 2009 Elsevier Ltd. All rights reserved.