COMPARATIVE CHARACTERIZATION OF NEW GLUTARYL 7-ACA AND CEPHALOSPORIN-C ACYLASES

被引:34
作者
ARAMORI, I
FUKAGAWA, M
TSUMURA, M
IWAMI, M
ONO, H
ISHITANI, Y
KOJO, H
KOHSAKA, M
UEDA, Y
IMANAKA, H
机构
[1] FUJISAWA PHARMACEUT CO LTD, EXPLORATORY RES LABS, 5-2-3 TOKODAI, TSUKUBA, IBARAKI 30026, JAPAN
[2] FUJISAWA PHARMACEUT CO LTD, MFG TECHNOL LABS, TSUKUBA, IBARAKI 30026, JAPAN
[3] FUJISAWA PHARMACEUT CO LTD, RES & DEV GRP, TSUKUBA, IBARAKI 30026, JAPAN
来源
JOURNAL OF FERMENTATION AND BIOENGINEERING | 1992年 / 73卷 / 03期
关键词
D O I
10.1016/0922-338X(92)90158-Q
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We performed a comparative characterization of three new cephalosporin acylases which were prepared from E. coli recombinant strains and found originally from Pseudomonas sp. A14, Bacillus laterosporus J1 and Pseudomonas diminuta N176. Both A14 and N176 acylases consisted of two non-identical subunits (alpha, beta) whose molecular weights were 28,000 (alpha), 61,000 (beta) and 26,000 (alpha), 58,000 (beta), respectively, whereas J1 acylase consisted of a single peptide with molecular weight of 70,000. The maximum specific activities of A14, J1 and N176 acylases for glutaryl 7-ACA were 7.1, 5.3 and 100 units/mg, respectively, and that of N176 acylase for cephalosporin C was 3.1 units/mg. The K(m) values of glutaryl 7-ACA for A14, J1 and N176 acylases were 2.1, 3.2 and 2.6 mM, respectively, and that of cephalosporin C for N176 acylase was 4.8 mM. A14, J1 and N176 acylases exhibited differential activities for cephalosporins having an aliphatic dicarboxylic acid in the acyl side chain and only N176 acylase showed an activity for cephalosporin C. N176 acylase as well as A14 acylase also showed a weak activity for a cephalosporin derivative having a heterocyclic carboxylic acid in the side chain. A14, J1 and N176 acylases catalyzed the reverse reaction to synthesize glutaryl 7-ACA from 7-ACA and glutaric acid, although the rate of the synthesis was 10 to 10(5) fold slower than that of hydrolysis. The activities of the cephalosporin acylases were considerably inhibited by the reaction products, 7-ACA and glutaric acid. The types of the inhibition by 7-ACA and glutaric acid were both competitive. A14, J1 and N176 acylases were thermostable, their residual activities exceeding more than 90% after treatment at 50-degrees-C for 1 h at their optimal pHs.
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页码:185 / 192
页数:8
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