Guo Wenjing, Tang Hui, Zhang Liping
Key Discipline of Biological Engineering of Hebei Province, College of Life Sciences, Hebei University, Baoding, China; Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, China.
J Basic Microbiol. 2014 Oct;54(10):1053-61. doi: 10.1002/jobm.201300920. Epub 2014 Mar 27.
crtYB, encoding lycopene cyclase and phytoene synthase was cloned from Rhodosporidium diobovatum ATCC 2527 by rapid amplification of cDNA ends method. The full-length cDNA of crtYB is 2, 330 bp and contains eight introns. The gene products is a 594 amino acids, with a predicted molecular mass of 65.63 kDa and a pI of 6.73. The N-terminus of the protein contains six transmembrane regions, which has been characterized as a lycopene beta-cyclase. The C-terminal half has squalene and phytoene synthase signatures that identified as phytoene synthetase. By heterologous complementary detection of this gene in E. coli and HPLC analysis, the regions responsible for phytoene synthesis and lycopene cyclization were localized within the protein.
通过cDNA末端快速扩增方法，从倒卵形红冬孢酵母ATCC 2527中克隆了编码番茄红素环化酶和八氢番茄红素合酶的crtYB。crtYB的全长cDNA为2330 bp，包含8个内含子。该基因产物为594个氨基酸，预测分子量为65.63 kDa，等电点为6.73。该蛋白质的N端包含6个跨膜区域，已被鉴定为番茄红素β-环化酶。C端的一半具有被鉴定为八氢番茄红素合酶的角鲨烯和八氢番茄红素合酶特征。通过在大肠杆菌中对该基因进行异源互补检测和高效液相色谱分析，确定了负责八氢番茄红素合成和番茄红素环化的区域位于该蛋白质内。
