José M Luengo

José M Luengo
Universidad de León | UNILEON · Department of Biochemistry and Molecular Biology

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150
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Introduction

Publications

Publications (150)
Article
Full-text available
The highly xerotolerant bacterium classified as Exiguobacterium sp. Helios isolated from a solar panel in Spain showed a close relationship to Exiguobacterium sibiricum 255-15 isolated from Siberian permafrost. Xerotolerance has not been previously described as a characteristic of the extremely diverse Exiguobacterium genus, but both strains Helios...
Chapter
Most bacteria belonging to the genus Pseudomonas can accumulate medium-chain poly-3-hydroxy-alkanoates (mcl-PHA) as storage materials. For this reason, these bacteria have a well-preserved enzymatic mechanism which involves the participation of a polymerization system (consisting of two different polymerases, PhaC1 and PhaC2), two proteins needed f...
Article
Biogenic amines (2‐phenylethylamine, tyramine, dopamine, epinephrine, norepinephrine, octopamine, histamine, tryptamine, serotonin, agmatine, cadaverine, putrescine, spermidine, spermine and certain aliphatic amines) are widely distributed organic molecules that play basic physiological functions in animals, plants and microorganisms. Pseudomonas s...
Article
Full-text available
Steroids are perhydro-1,2-cyclopentanophenanthrene derivatives that are almost exclusively synthesised by eukaryotic organisms. Since the start of the Anthropocene, the presence of these molecules, as well as related synthetic compounds (ethinylestradiol, dexamethasone, and others), has increased in different habitats due to farm and municipal effl...
Article
Full-text available
Hydroxylation of steroids has acquired special relevance for the pharmaceutical industries. Particularly, the 11β‐hydroxylation of steroids is a reaction of biotechnological importance currently carried out at industrial scale by the fungus Cochliobolus lunatus. In this work, we have identified the genes encoding the cytochrome CYP103168 and the re...
Article
In this study, the catabolic pathway required for the degradation of the biogenic amine histamine (Hin) was genetically and biochemically characterized in Pseudomonas putida U. The 11 proteins (HinABCDGHFLIJK) that participate in this pathway are encoded by genes belonging to three loci hin1, hin2 and hin3 and by the gene hinK. The enzymes HinABCD...
Chapter
The study of the catabolic potential of microbial species isolated from different habitats has allowed the identification and characterization of bacteria able to assimilate bile acids and other steroids (e.g., testosterone and 4-androsten-3,17-dione). From soil samples, we have isolated several strains belonging to genus Pseudomonas that grow effi...
Article
Pseudomonas putida N, a poly-3-hydroxyalkonate (PHA)-producing bacterium, showing ampicillin resistance, is an unusual strain. In the presence of this antibiotic, it grows as giant cells (25-50μm) forming complex networks inter-connected by micro-tubular structures. The transformation of this bacterium with a plasmid containing the gene phaF, which...
Article
The polyhydroxyalkanoate (PHA)-overproducing mutant Pseudomonas putida U fadBA (Pp�fadBA), lacks the genes encoding the main β-oxidation pathway (FadBA). This strain accumulates enormous amounts of bioplastics when cultured in chemically defined media containing PHA-precursors (different n-alkanoic or n-aryl-alkanoic acids) and an additional carbon...
Article
Pseudomonas putida DOC21, a soil-dwelling proteobacterium, catabolises a variety of steroids and bile acids. Transposon mutagenesis and bioinformatics analyses identified four clusters of steroid degradation (std) genes encoding a single catabolic pathway. The latter includes three predicted acyl-CoA synthetases encoded by stdA1, stdA2 and stdA3, r...
Article
Nineteen different steroid-degrading bacteria were isolated from soil samples by using selective media containing either cholesterol or deoxycholate as sole carbon source. Strains that assimilated cholesterol (17 COL strains) were gram-positive, belonging to the genera Gordonia, Tsukamurella, and Rhodococcus, and grew on media containing other ster...
Article
Degradation of tyramine and dopamine by Pseudomonas putida U involves the participation of twenty one proteins organized in two coupled catabolic pathways, Tyn (tynABFEC tynG tynR tynD, 12 338 bp) and Hpa (hpaR hpaBC hpaHI hpaX hpaG1G2EDF hpaA hpaY, 12 722 bp). The Tyn pathway catalyses the conversion of tyramine and dopamine into 4-hydroxyphenylac...
Chapter
Full-text available
Unusual polyhydroxyalkanoates (UnPHAs) constitute a particular group of polyoxo(thio)esters belonging to the PHA family, which are tailored with uncommon monomers. Thus, unusual PHAs include (1) polyhydroxyalkanoates (PHAs) of microbial origin that have been synthesized either from natural monomers bearing different chemical functions, or from chem...
Article
The substrate specificity of the two polymerases (PhaC1 and PhaC2) involved in the biosynthesis of medium-chain-length poly-hydroxyalkanoates (mcl PHAs) in Pseudomonas putida U has been studied in vivo. For these kind of experiments, two recombinant strains derived from a genetically engineered mutant in which the whole pha locus had been deleted (...
Article
In Pseudomonas putida U two different pathways (Pea, Ped) are required for the conversion of 2-phenylethylamine and 2-phenylethanol into phenylacetic acid. The 2-phenylethylamine pathway (PeaABCDEFGHR) catalyses the transport of this amine, its deamination to phenylacetaldehyde by a quinohaemoprotein amine dehydrogenase and the oxidation of this co...
Data
Secondary structure prediction for PhaC1 and PhaC2. PhaC1 and PhaC2 sequences are shown in red and black, whereas their corresponding secondary structure predictions are shown in pink and blue respectively. Numbering is according to the PhaC1 sequence. Regions with significant discrepancies on prediction are highlighted within a green box. The resi...
Article
Functional analyses of the different proteins involved in the synthesis and accumulation of polyhydroxyalkanoates (PHAs) in P. putida U were performed using a mutant in which the pha locus had been deleted (PpUDeltapha). These studies showed that: (i) Pha enzymes cannot be replaced by other proteins in this bacterium, (ii) the transformation of PpD...
Article
Full-text available
Polyhydroxyalkanoates (PHAs) can be catabolized by many microorganisms using intra- or extracellular PHA depolymerases. Most of our current knowledge of these intracellular enzyme-coding genes comes from the analysis of short chain length PHA depolymerases, whereas medium chain length PHA (mcl-PHA) intracellular depolymerization systems still remai...
Chapter
Polyhydroxyalkanoates (PHAs) are optically active biopolyoxoesters composed of (R)-3-hydroxy fatty acids which represent a complex class of storage polyesters. They are synthesized by some Archaea and by a wide range of Gram-positive and Gram-negative bacteria in aerobic and anaerobic environments. PHAs are accumulated as inclusions in the bacteria...
Chapter
One of the most important causes contributing to the modification of living conditions is the accumulation of many chemical compounds in broad areas of the planet. Among the thousands of new contaminants released to the environment every year, the appearance in many habitats (aquatic, aerial or terrestrial) of quite stable molecules containing arom...
Article
The gene (acs) encoding the acetyl-CoA synthetase (Acs) in Pseudomonas putida U has been cloned, sequenced and expressed in different microbes. The protein has been purified and characterized from a biochemical, structural and evolutionary point of view. Disruption or deletion of acs handicapped the bacterium for growth in a chemically defined medi...
Article
4-Hydroxyphenylacetic acid-3-hydroxylase from Pseudomonas putida U was purified to homogeneity (96-fold) from bacterial cultures grown in a chemically defined medium containing 4-hydroxyphenylacetic acid as the sole carbon source. The maximal rate of catalysis occurred at pH 7.5 and 40°C. Under these conditions, the Km values calculated for 4-hydro...
Article
Pseudomonas putida U grown in a chemically defined medium containing octanoic acid as the sole carbon source accumulated a homopolymer of poly(3-hydroxyoctanoate) as intracellular reserve material, and metabolized the polymer during the late exponential phase of growth. Kinetic measurement of the uptake of [1-14C]octanoic acid by cells at 34°C in 8...
Article
A genetically engineered strain of Pseudomonas putida U designed for the identification of new therapeutic herbicides has been obtained. In this bacterium, deletion of the homogentisate gene cluster (hmgRABC) confers upon this mutant huge biotechnological possibilities since it can be used: (i) as a target for testing new specific herbicides (p-hyd...
Article
The aroA gene of Yersinia ruckeri, which encodes 5-enolpyruvylshikimate 3-phosphate synthase, was insertionally inactivated with a DNA fragment containing a kanamycin resistance determinant and reintroduced by allelic exchange into the chromosome of Y. ruckeri 21102 O1 by means of the suicide vector pIVET8. The Y. ruckeri aroA::Kan(r) mutant was hi...
Article
Full-text available
The complete catabolic pathway involved in the assimilation of 3-hydroxyphenylacetic acid (3-OH-PhAc) in Pseudomonas putida U has been established. This pathway is integrated by the following: (i) a specific route (upper pathway), which catalyzes the conversion of 3-OH-PhAc into 2,5-dihydroxyphenylacetic acid (2,5-diOH-PhAc) (homogentisic acid, Hmg...
Article
Overexpression of the gene encoding the poly-3-hydroxy-n-phenylalkanoate (PHPhA) depolymerase (phaZ) in Pseudomonas putida U avoids the accumulation of these polymers as storage granules. In this recombinant strain, the 3-OH-acyl-CoA derivatives released from the different aliphatic or aromatic poly-3-hydroxyalkanoates (PHAs) are catabolized throug...
Chapter
The phenylacetyl-CoA catabolon is a complex degradative unit integrated by several catabolic pathways that transform different unrelated aromatic compounds (i.e., phenylacetic acid [PhAc], phenylacetaldehyde, styrene, tropic acid, 2-phenylethylamine, 2-phenylethanol, ethylbenzene, penylacetylamides and other esters, n-phenylalkanoates with an even...
Article
Full-text available
We report an easy procedure for isolating chromosome-clustered genes. By following this methodology, the entire set of genes belonging to the phenylacetic acid (PhAc; 18-kb) pathway as well as those required for the synthesis and mobilization of different polyhydroxyalkanoates (PHAs; 6.4 kb) in Pseudomonas putida U were recovered directly from the...
Article
Full-text available
Pseudomonas putida metabolizes Phe and Tyr through a peripheral pathway involving hydroxylation of Phe to Tyr (PhhAB), conversion of Tyr into 4-hydroxyphenylpyruvate (TyrB), and formation of homogentisate (Hpd) as the central intermediate. Homogentisate is then catabolized by a central catabolic pathway that involves three enzymes, homogentisate di...
Article
Full-text available
The term 'biomaterials' includes chemically unrelated products that are synthesised by microorganisms (or part of them) under different environmental conditions. One important family of biomaterials is bioplastics. These are polyesters that are widely distributed in nature and accumulate intracellularly in microorganisms in the form of storage gran...
Article
I propose a hypothesis, called the systems convergence-divergence hypothesis, which attempts to explain how the existence and evolution of supraindividual systems (social, political or economic) might be predicted from an analysis of the basic structures that make up their individual entities (molecules and cells) and how, reciprocally, study of th...
Article
Yersinia ruckeri possesses a quorum-sensing system detected by cross-streaking against the white mutant Chromobacterium voilaceum CV0blu. Quorum sensing, which occurs in a number of Gram-negative pathogens, is known to control virulence gene expression through cell to cell communication. There are two genes required for quorum sensing which are lux...
Article
New bioplastics containing aromatic or mixtures of aliphatic and aromatic monomers have been obtained using genetically engineered strains of Pseudomonas putida. The mutation (-) or deletion (Delta) of some of the genes involved in the beta-oxidation pathway (fadA(-), fadB(-) Delta fadA or Delta fad BA mutants) elicits a strong intracellular accumu...
Article
Oligonucleotide primers specific for the Staphylococcus aureus gap gene were previously designed to identify 12Staphylococcus spp. by PCR. In the present study,AluI digestion of PCR-generated products rendered distinctive restriction fragment length polymorphism patterns that allowed 24 Staphylococcus spp. to be identified with high specificity.
Article
The aroA gene of Yersinia ruckeri, which encodes 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase was cloned by complementation of the aroA mutation in Escherichia coli AB2829 by using pUC18 plasmid as a vector. Nucleotide sequence of the aroA gene revealed an open reading frame of 427 amino acids showing a high degree of homology to other bacter...
Article
The term catabolon was introduced to define a complex functional unit integrated by different catabolic pathways, which are, or could be, co-ordinately regulated, and that catalyses the transformation of structurally related compounds into a common catabolite. The phenylacetyl-CoA catabolon encompasses all the routes involved in the transformation...
Article
New poly(β-hydroxyalkanoates) having aromatics groups (so-called PHPhAs) from a microbial origin have been characterized. These polymers were produced and accumulated as reserve materials when a β-oxidation mutant of Pseudomonas putida U, disrupted in the gene that encodes the 3-ketoacyl−CoA thiolase (fadA), was cultured in a chemically defined med...
Article
In Pseudomonas putida U, the degradation of n-alkanoic and n-phenylalkanoic acids is carried out by two sets of beta-oxidation enzymes (betaI and betaII). Whereas the first one (called betaI) is constitutive and catalyses the degradation of n-alkanoic and n-phenylalkanoic acids very efficiently, the other one (betaII), which is only expressed when...
Article
In Pseudomonas putida U, the degradation of n-alkanoic and n-phenylalkanoic acids is carried out by two sets of β-oxidation enzymes (βI and βII). Whereas the first one (called βI) is constitutive and catalyses the degradation of n-alkanoic and n-phenylalkanoic acids very efficiently, the other one (βII), which is only expressed when some of the gen...
Article
The gap gene of Staphylococcus aureus, encoding glyceraldehyde-3-phosphate dehydrogenase, was used as a target to amplify a 933-bp DNA fragment by PCR with a pair of primers 26 and 25 nucleotides in length. PCR products, detected by agarose gel electrophoresis, were also amplified from 12 Staphylococcus spp. analyzed previously. Hybridization with...
Article
Full-text available
A new class of glutamate dehydrogenase (GDH) is reported. The GDH of Streptomyces clavuligerus was purified to homogeneity and characterized. It has a native molecular mass of 1,100 kDa and exists as an alpha(6) oligomeric structure composed of 183-kDa subunits. GDH, which requires AMP as an essential activator, shows a maximal rate of catalysis in...
Article
Full-text available
Referencia OEPM: P9801503.-- Fecha de solicitud: 14/07/1998.-- Titular: Consejo Superior de Investigaciones Científicas (CSIC), Universidad de León. La presente invención consiste en la realización de un procedimiento para producir ácido 2-hidroxifenil-acético (2HPA) en Escherichia coli. La invención reivindica el uso los marcos de lectura abiertos...
Article
Full-text available
Aerobic degradation of phenylacetic acid in Pseudomonas putida U is carried out by a central catabolism pathway (phenylacetyl-coenzyme A [CoA] catabolon core). Induction of this route was analyzed by using different mutants specifically designed for this objective. Our results revealed that the true inducer molecule is phenylacetyl-CoA and not othe...