Detail of "9046-67-7"

Reference

Primary structure of the Streptomyces R61 extracellular DD-peptidase

Primary structure of the Streptomyces R61 extracellular DD-peptidase. 1. Cloning into Streptomyces lividans and nucleotide sequence of the gene. Duez, Colette; Piron-Fraipont, Claudine; Joris, Bernard; Dusart, Jean; Urdea, Mickey S.; Martial, Joseph A.; Frere, Jean Marie; Ghuysen, Jean Marie (Fac. Med., Univ. Liege, Liege, Belg.). Eur. J. Biochem., 162(3), 509-18 (English) 1987. CODEN: EJBCAI. ISSN: 0014-2956. DOCUMENT TYPE: Journal CA Section: 3 (Biochemical Genetics) An 11,450-base DNA fragment contg. the gene for the extracellular active-site serine DD-peptidase [9046-67-7] of Streptomyces R61 was cloned in S. lividans using the high-copy-no. plasmid pIJ702 as vector. Amplified expression of the excreted enzyme was obsd. Producing clones were identified with the help of a specific antiserum directed against the pure DD-peptidase. The coding sequence of the gene was then located by hybridization with a specific nucleotide probe and sub-fragments were obtained from which the nucleotide sequence of the structural gene and the putative promoter and terminator regions were detd. The sequence suggests that the gene codes for a 406-amino-acid precursor. When compared with the excreted mature DD-peptidase, this precursor possesses a cleavable 31-amino-acid N-terminal extension which has the characteristics of a signal peptide, and a cleavable 26-amino-acid C-terminal extension. On the basis of the data of B. 37228-77-6 and 107761-92-2 are also in the experiment. Joris et al. (1987), the open reading frame coding for the synthesis of the DD-peptidase was established. Comparison of the primary structure of the Streptomyces R61 DD-peptidase with those of several active-site serine b-lactamases [9073-60-3] and penicillin-binding proteins of Escherichia coli shows homol. in those sequences that comprise the active-site serine residue. When the comparison is broadened to the complete amino acid sequences, significant homol. is obsd. only for the pair Streptomyces R61 DD-peptidase/E. coli ampC b-lactamase (class C). Since the Streptomyces R61 DD-peptidase and b-lactamases of class A have very similar 3-dimensional structures, it is concluded that these tertiary features are probably also shared by the b-lactamases of class C, i.e. that the Streptomyces R61 DD-peptidase and the b-lactamases of classes A and C are related in an evolutionary sense. .

Semisynthesis of human insulin utilizing chemically modified carboxypeptidase Y

Semisynthesis of human insulin utilizing chemically modified carboxypeptidase Y. Breddam, Klaus; Johansen, Jack T. (Chem. Dep., Carlsberg Lab., Copenhagen Valby DK-2500, Den.). Carlsberg Res. Commun., 49(4), 463-72 (English) 1984. CODEN: CRCODS. ISSN: 0105-1938. DOCUMENT TYPE: Journal CA Section: 16 (Fermentation and Bioindustrial Chemistry) Section cross-reference(s): 2, 63 The C-terminal alanyl residue in the B-chain of porcine insulin [9004-10-8] can be exchanged with a threonyl residue in a carboxypeptidase Y (CPD-Y) [9046-67-7] catalyzed transpeptidation reaction using threonine amide as nucleophile. However, with this procedure the transpeptidation product, human insulin amide, was obtained in relatively low yields. On the other hand, Hg halide derivs. of CPD-Y catalyze the transpeptidation reaction with porcine insulin effectively, and human insulin amide can be isolated in high yield. Deamidation of human insulin amide to yield essentially homogeneous human insulin can be achieved using CPD-Y modified with methyl mercuric iodide [143-36-2].