Enzyme participating in c-terminal amidation, and method of preparing same and use thereof
申请人:SHISEIDO COMPANY LIMITED
公开号:EP0666318A1
公开(公告)日:1995-08-09
An enzyme catalyzing the conversion of a peptide C-terminal glycine adduct represented by the following formula (I):
(wherein A represents a residue other than α-amino group or imino group and α-carboxylic group derived from naturally occurring α-amino acid, X represents hydrogen atom or a residue of an amino acid derivative which is bonded to N atom through carbonyl group) to a peptide C-terminal α-hydroxyglycine adduct represented by the following formula (II):
(wherein A and X have the same meanings as above), and further catalyzing the conversion of the peptide C-terminal α-hydroxylglycine adduct represented by the above formula (II) to a C-terminal amidated compound represented by the following formula (III):
(wherein A and X have the see meanings as defined above), which comprises culturing host cells transformed with an expression plasmid comprising a DNA coding for said enzyme, and recovering both or either of said enzymes from the culture, characterized in that in said DNA a portion corresponding to the membrane spanning region has been excised.
催化下式(I)所代表的肽 C 端甘氨酸加合物转化的酶:
(其中 A 代表α-氨基或亚胺基和α-羧基以外的残基,α-氨基或亚胺基和α-羧基来自天然存在的α-氨基酸,X 代表氢原子或通过羰基与 N 原子结合的氨基酸衍生物残基)转化为下式(II)代表的肽 C-末端α-羟基甘氨酸加合物:
(其中A和X的含义同上),并进一步催化由上式(II)代表的肽C-末端α-羟基甘氨酸加合物转化为由下式(III)代表的C-末端酰胺化化合物:
(其中A和X的含义与上式相同),其中包括培养用包含编码上述酶的DNA的表达质粒转化的宿主细胞,并从培养物中回收上述两种或其中一种酶,其特征在于,在上述DNA中,对应于跨膜区的部分已被切除。