Cloning and nucleotide sequencing of new glutaryl 7-ACA and cephalosporin C acylase genes from Pseudomonas strains

Characteristic of immobilized cephalosporin C acylase and its application in one-step enzymatic conversion of cephalosporin C to 7-aminocephalosporanic acid

“…CPC acylase has been found in several microorganisms, such as Pseudomonas sp., Aeromonas sp., Arthrobacter viscous etc., and some CPC acylase genes have been sequenced, cloned and studied. Unfortunately, these CPC acylases exhibit low specificity and activity towards CPC and could not meet the requisite production efficiency (Aramori et al 1991;Saito et al 1996;Deshpande et al 1996;Sonawane 2006). Extensive attempts have been made to improve the specific activity of those CPC acylases, and some enzymes evolved with high CPC specificity represent a hallmark for industrial production of 7-ACA (Pollegioni et al 2005).…”

Site-Directed Mutagenesis of Cephalosporin C Acylase and Enzymatic Conversion of Cephalosporin C to 7-Aminocephalosporanic Acid

“…The NH groups from the backbone of His70β and side chain of Asn242β formed the oxyanion hole for carboxyl group on CPC. For the binding sites, oxygen atoms from the Cephalosporin C Acylase in N176 [11] 3. carboxylate group of CPC interacted with Arg24β, Tyr32β and His57β. The amino adipyl moiety of CPC was stabilized by the formed hydrogen bond with His178β which simultaneously interacted with Asp177β stated by Figure 4.…”

Characterization of an industrial biocatalyst: Immobilized glutaryl-7-ACA acylase

“…Unsuccessful attempts to obtain a direct biocatalytic transformation of 1 into 2 with an acceptable conversion rate (Aramori et al, 1991) have led to the development of the two-step enzymatic process shown in Scheme 1 (Shibuya et al, 1981;Tischer et al, 1992). This route includes the oxidative deamination of CefC to 7-␤-(4-carboxybutanamido) cephalosporanic acid (3, glutaryl-7-ACA) catalyzed by a D-amino acid oxidase (DAAO) (Sauber, 1993;Pollegioni et al, 1993;Andreesen, 1993, 1996), and the subsequent deacylation of 3 catalyzed by a glutaryl-7-ACA acylase (GA).…”