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An acidic glutaryl‐7‐aminocephalosporanic acid acylase from Pseudomonasnitroreducens
Author(s) -
Lee YunHuey,
Chang TeSheng,
Liu HonJu,
Chu WenShen
Publication year - 1998
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1998.tb00520.x
Subject(s) - isoelectric point , molecular mass , enzyme , chemistry , chromatography , isoelectric focusing , biochemistry , sepharose , enzyme assay , yield (engineering) , specific activity , osmotic shock , materials science , gene , metallurgy
A glutaryl‐7‐aminocephalosporanic acid (GL‐7‐ACA) acylase was purified 58‐fold from Pseudomonas nitroreducens in a two‐step procedure involving osmotic shock and carboxymethyl‐Sepharose chromatography with a yield of 26%. The molecular mass of the native enzyme was 58 kDa. SDS/PAGE revealed that it consisted of two non‐identical subunits with molecular masses of 35 and 21 kDa. The isoelectric point of the purified enzyme was 5.3. The enzyme had an optimal pH of 5.5 and an optimal temperature of 43 °C. The purified enzyme exhibited not only GL‐7‐ACA acylase activity but also γ‐glutamyltranspeptidase activity. The K m values of the enzyme for GL‐7‐ACA and L‐γ‐glutamyl p ‐nitroanilide were 10.41 mM and 5.92 μM respectively.