HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Miñambres, B. Articles by Luengo, J. M. Search for Related Content PubMed PubMed Citation Articles by Miñambres, B. Articles by Luengo, J. M. Social Bookmarking                      What's this?

Volume 271, Number 52, Issue of December 27, 1996 pp. 33531-33538
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Molecular Cloning and Expression in Different Microbes of the DNA Encoding Pseudomonas putida U Phenylacetyl-CoA Ligase
USE OF THIS GENE TO IMPROVE THE RATE OF BENZYLPENICILLIN BIOSYNTHESIS IN PENICILLIUM CHRYSOGENUM

(Received for publication, July 9, 1996, and in revised form, September 27, 1996)

From the Departamento de Bioquímica y Biología Molecular, Facultades de Veterinaria y Biología, Universidad de León, 24007, León, España and ^§ Antibióticos S.A. Leon, León, España

The gene encoding phenylacetyl-CoA ligase (pcl), the first enzyme of the pathway involved in the aerobic catabolism of phenylacetic acid in Pseudomonas putida U, has been cloned, sequenced, and expressed in two different microbes. In both, the primary structure of the protein was studied, and after genetic manipulation, different recombinant proteins were analyzed. The pcl gene, which was isolated from P. putida U by mutagenesis with the transposon Tn5, encodes a 48-kDa protein corresponding to the phenylacetyl-CoA ligase previously purified by us (Martínez-Blanco, H., Reglero, A. Rodríguez-Aparicio, L. B., and Luengo, J. M. (1990) J. Biol. Chem. 265, 7084-7090). Expression of the pcl gene in Escherichia coli leads to the appearance of this enzymatic activity, and cloning and expression of a 10.5-kb DNA fragment containing this gene confer this bacterium with the ability to grow in chemically defined medium containing phenylacetic acid as the sole carbon source. The appearance of phenylacetyl-CoA ligase activity in all of the strains of the fungus Penicillium chrysogenum transformed with a construction bearing this gene was directly related to a significant increase in the quantities of benzylpenicillin accumulated in the broths (between 1.8- and 2.2-fold higher), indicating that expression of this bacterial gene (pcl) helps to increase the pool of a direct biosynthetic precursor, phenylacetyl-CoA. This report describes the sequence of a phenylacetyl-CoA ligase for the first time and provides direct evidence that the expression in P. chrysogenum of a heterologous protein (involved in the catabolism of a penicillin precursor) is a useful strategy for improving the biosynthetic machinery of this fungus.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				E. Arias-Barrau, A. Sandoval, G. Naharro, E. R. Olivera, and J. M. Luengo A Two-component Hydroxylase Involved in the Assimilation of 3-Hydroxyphenyl Acetate in Pseudomonas putida J. Biol. Chem., July 15, 2005; 280(28): 26435 - 26447. [Abstract] [Full Text] [PDF]

				B. Garcia, E. R. Olivera, A. Sandoval, E. Arias-Barrau, S. Arias, G. Naharro, and J. M. Luengo Strategy for Cloning Large Gene Assemblages as Illustrated Using the Phenylacetate and Polyhydroxyalkanoate Gene Clusters Appl. Envir. Microbiol., August 1, 2004; 70(8): 5019 - 5025. [Abstract] [Full Text] [PDF]

				E. Diaz, A. Ferrandez, M. A. Prieto, and J. L. Garcia Biodegradation of Aromatic Compounds by Escherichia coli Microbiol. Mol. Biol. Rev., December 1, 2001; 65(4): 523 - 569. [Abstract] [Full Text] [PDF]

				B. García, E. R. Olivera, B. Miñambres, D. Carnicero, C. Muñiz, G. Naharro, and J. M. Luengo Phenylacetyl-Coenzyme A Is the True Inducer of the Phenylacetic Acid Catabolism Pathway in Pseudomonas putida U Appl. Envir. Microbiol., October 1, 2000; 66(10): 4575 - 4578. [Abstract] [Full Text]

				B. Garcia, E. R. Olivera, B. Minambres, M. Fernandez-Valverde, L. M. Canedo, M. A. Prieto, J. L. Garcia, M. Martinez, and J. M. Luengo Novel Biodegradable Aromatic Plastics from a Bacterial Source. GENETIC AND BIOCHEMICAL STUDIES ON A ROUTE OF THE PHENYLACETYL-CoA CATABOLON J. Biol. Chem., October 8, 1999; 274(41): 29228 - 29241. [Abstract] [Full Text] [PDF]

				J. M. Mingot, M. A. Penalva, and J. M. Fernandez-Canon Disruption of phacA, an Aspergillus nidulans Gene Encoding a Novel Cytochrome P450 Monooxygenase Catalyzing Phenylacetate 2-Hydroxylation, Results in Penicillin Overproduction J. Biol. Chem., May 21, 1999; 274(21): 14545 - 14550. [Abstract] [Full Text] [PDF]

				A. Ferrandez, B. Minambres, B. Garcia, E. R. Olivera, J. M. Luengo, J. L. Garcia, and E. Diaz Catabolism of Phenylacetic Acid in Escherichia coli. CHARACTERIZATION OF A NEW AEROBIC HYBRID PATHWAY J. Biol. Chem., October 2, 1998; 273(40): 25974 - 25986. [Abstract] [Full Text] [PDF]

				E. R. Olivera, B. Minambres, B. Garcia, C. Muniz, M. A. Moreno, A. Ferrandez, E. Diaz, J. L. Garcia, and J. M. Luengo Molecular characterization of the phenylacetic acid catabolic pathway in Pseudomonas putida U: The phenylacetyl-CoA catabolon PNAS, May 26, 1998; 95(11): 6419 - 6424. [Abstract] [Full Text] [PDF]

				A. Velasco, S. Alonso, J. L. García, J. Perera, and E. Díaz Genetic and Functional Analysis of the Styrene Catabolic Cluster of Pseudomonas sp. Strain Y2 J. Bacteriol., March 1, 1998; 180(5): 1063 - 1071. [Abstract] [Full Text]