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

Monti, Daniela; Carrea, Giacomo; Riva, Sergio; Baldaro, Eva; Frare, Giovanni

doi:10.1002/1097-0290(20001020)70:2<239::aid-bit13>3.0.co;2-i

Cited by 44 publications

(19 citation statements)

References 14 publications

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“…A co-integration with the cefG gene of A. chrysogenum encoding the DAC acyltransferase resulted in the conversion of adipoyl-7-ADAC into adipoyl-7-ACA (Crawford et al, 1995). Likewise, an in vitro conversion of adipoyl-7-ACA with a glutaryl-7-ACA-acylase yields 7-ACA (Monti et al, 2000) which is a preferred precursor for cephalosporin biosynthesis. Recently, the expression of the A. chrysogenum genes cefD1, cefD2, cefEF and cefG in a P. chrysogenum strain lacking the isopenicillin N-acyltransferase has lead to significant amounts of intracellular produced deacetylcephalosporin C and cephalosporin C (Ullan et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Expression of the transporter encoded by the cefT gene of Acremonium chrysogenum increases cephalosporin production in Penicillium chrysogenum

Nijland

Kovalchuk

Berg

et al. 2008

Fungal Genetics and Biology

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Section: Introductionmentioning

confidence: 99%

Expression of the transporter encoded by the cefT gene of Acremonium chrysogenum increases cephalosporin production in Penicillium chrysogenum

Nijland

Kovalchuk

Berg

et al. 2008

Fungal Genetics and Biology

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“…Because of the role of the two-step enzymatic process in the 7-ACA industry, much attention has been paid to the immobilization of d-amino acid oxidase and GL-7-ACA acylase. Among these immobilization techniques, covalent attachment has been successful in reinforcing the combining power and enhancing the stability of these enzymes (Golini et al 1995;Monti et al 2000;Zheng et al2006;Lim et al 2006). Contrarily, there have been few studies on the immobilization of CPC acylase and its application (Pollegioni et al 2005).…”

mentioning

confidence: 99%

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

Zhu

Luo

Chang

et al. 2010

World J Microbiol Biotechnol

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Cephalosporin C (CPC) acylase is an enzyme which hydrolyzes CPC to 7-aminocephalosporanic acid (7-ACA) directly, and therefore has great potential in industrial application. In this study, the CPC acylase from a recombinant Escherichia coli was purified to high purity by immobilized metal affinity chromatography, and the CPC acylase was covalently attached to three kinds of epoxy supports, BB-2, ES-V-1 and LX-1000EP. The immobilized CPC acylase with LX-1000EP as the support shows the highest activity (81 U g -1 ) suggesting its potential in industrial 7-ACA production. The activity of immobilized enzyme was found to be optimal at pH between 8.5 and 9.5 and to increase with temperature elevation until 55°C. Immobilized CPC acylase showed good stability at pH between 8.0 and 9.5 and at temperature up to 40°C. To avoid product degradation, the production of 7-ACA utilizing immobilized enzyme was carried out at 25°C, pH 8.5 in a designed reactor. Under optimal reaction conditions, a very high 7-ACA yield of 96.7% was obtained within 60 min. In the results of repeated batch production of 7-ACA, 50% activity of the initial cycle was maintained after being recycled 24 times and the average conversion rate of CPC reached 98%.

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“…One unit of DAAO activity corresponded to the formation of 1 lmol min -1 of pyruvate at 37°C. The activity of immobilized GA was measured following the method described by Monti (Monti et al 2002). One unit of GA activity was defined as the amount of enzyme which produced 1 lmol min -1 of 7-ACA at 37°C.…”

Section: Enzyme Assaymentioning

confidence: 99%

Single-pot conversion of cephalosporin C to 7-aminocephalosporanic acid in the absence of hydrogen peroxide

Tan

Zhang

Song

et al. 2009

World J Microbiol Biotechnol

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In this study, D-amino acid oxidase (DAAO) and catalase (CAT) in the permeabilized recombinant Pichia pastori cells were well investigated. It appeared that their thermal stability was negatively correlated with the apparent enzymatic activities. The frozen-melted cells presented the best stability and the lowest apparent activities of DAAO and CAT, whereas the cetyltrimethylammonium bromide (CTAB) permeabilized cells displayed the weakest stability and the highest apparent activities of the two enzymes. Simultaneous action of DAAO and CAT in the CTAB-permeabilized cells and glutaryl-7-aminocephalosporanic acid acylase (GA) immobilized on carrier contributed to the conversion of cephalosporin C (CPC) to 7-aminocephalosporanic acid (7-ACA) with a yield of 76.2%. During such a reaction cycle, no visible activity loss occurred at the immobilized GA, whereas the loss rates of DAAO and CAT activities were about 0.029 and 1.13 U min -1 , respectively. Nevertheless, this problem could be easily solved by continuous feeding of the new permeabilized cell suspension at the rate of 6 ml h -1 to the reactor. Following such a fed-batch strategy, these permeabilized cells and the immobilized GA could be efficiently reused for 6 and 15 reaction cycles, respectively, yielding around 76% 7-ACA at each reaction cycle.

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

Cited by 44 publications

References 14 publications

Expression of the transporter encoded by the cefT gene of Acremonium chrysogenum increases cephalosporin production in Penicillium chrysogenum

Expression of the transporter encoded by the cefT gene of Acremonium chrysogenum increases cephalosporin production in Penicillium chrysogenum

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

Single-pot conversion of cephalosporin C to 7-aminocephalosporanic acid in the absence of hydrogen peroxide

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