公开(公告)号：US06472196B1

公开(公告)日：2002-10-29

申请号：US08214018

申请日：1994-03-15

申请人： Nobuhiro Fukuhara ,  Setsuo Yoshino ,  Kaoru Yamamoto ,  Tomoyuki Se ,  Satori Sone ,  Yoshiyuki Nakajima ,  Maki Suzuki ,  Nobuyoshi Makiguchi

发明人： Nobuhiro Fukuhara ,  Setsuo Yoshino ,  Kaoru Yamamoto ,  Tomoyuki Se ,  Satori Sone ,  Yoshiyuki Nakajima ,  Maki Suzuki ,  Nobuyoshi Makiguchi

IPC分类号： C12N988

CPC分类号： C12P13/222 ,  C12N9/88

摘要： In order to provide technical information necessary for the production of L-phenylalanine ammonia-lyase (PAL) by utilizing genetic engineering techniques, the structural gene for PAL and the amino acid sequence of PAL have been elucidated in Rhodosporidium toruloides, and novel recombinant DNA plasmids (e.g., pSW101, pYtrp6 and pKY201) have been created by inserting a DNA strand coding for the PAL gene between the 3′-terminus of the promoter region and the 5′-terminus of the terminator region. Moreover, transformants having such a novel recombinant DNA plasmid have been created, and a process for the production of PAL by growing such a novel transformant so as to cause PAL to be produced and accumulated in the culture has been established. Furthermore, there has been established a novel technique for the production of L-phenylalanine by reacting an ammonia donor with cinnamic acid in the presence of the PAL prepared by the aforesaid novel process.

摘要翻译： 为了提供利用遗传工程技术生产L-苯丙氨酸氨裂解酶（PAL）所必需的技术信息，已经在Rhodosporidium toruloides中阐明了PAL的结构基因和PAL的氨基酸序列，新的重组DNA质粒 （例如，pSW101，pYtrp6和pKY201）是通过将编码PAL基因的DNA链插入到启动子区域的3'端和终止区域的5'末端而产生的。此外，具有这样一种新颖的转化体 已经建立了重组DNA质粒，并且通过生长这样一种新的转化体以使培养中产生和积累PAL来生产PAL的方法已被建立。此外，已经建立了一种生产新技术 在通过上述新方法制备的PAL存在下，使氨供体与肉桂酸反应，生成L-苯丙氨酸。