Enrica Galli

Effect on wheat root development of inoculation with an Azospirillum brasilense mutant with altered indole-3-acetic acid production

To evaluate the involvement of indole-3-acetic acid (IAA) in promotion of root development, the effects of inoculation of wheat seedlings with Azospirillum brasilense SpM7918, a very low-IAA producer, were estimated. Compared with the wild-type strain Sp6, SpM7918 showed a reduced ability to promote root system development in terms both of number and length of lateral roots and of distribution of root hairs.

Styrene Catabolism by a Strain of Pseudomonas fluorescens

A strain of Pseudomonas fluorescens, capable of growing on styrene as the sole C source was isolated from enrichment cultures. From the Pseudomonas cultures supplied with styrene, phenylacetic and o-hydroxyphenylacetic acids were isolated and identified, o-Hydroxyphenylacetic acid was also isolated from the cultures supplied with phenylacetate. Homogentisate 1,2-dioxygenase is induced in the cells grown either on styrene or phenylacetate or o-hydroxyphenylacetate. A pathway for styrene metabolism through the intermediary formation of phenylacetate and o-hydroxyphenylacetate which is further oxidized via homogentisate is suggested and discussed.

Organization and Regulation of meta Cleavage Pathway Genes for Toluene and o-Xylene Derivative Degradation in Pseudomonas stutzeri OX1

ABSTRACT Pseudomonas stutzeri OX1 meta pathway genes for toluene and o-xylene catabolism were analyzed, and loci encoding phenol hydroxylase, catechol 2,3-dioxygenase, 2-hydroxymuconate semialdehyde dehydrogenase, and 2-hydroxymuconate semialdehyde hydrolase were mapped. Phenol hydroxylase converted a broad range of substrates, as it was also able to transform the nongrowth substrates 2,4-dimethylphenol and 2,5-dimethylphenol into 3,5-dimethylcatechol and 3,6-dimethylcatechol, respectively, which, however, were not cleaved by catechol 2,3-dioxygenase. The identified gene cluster displayed a gene order similar to that of thePseudomonas sp. strain CF600 dmp operon for phenol catabolism and was found to be coregulated by thetou operon activator TouR. A hypothesis about the evolution of the toluene and o-xylene catabolic pathway inP. stutzeri OX1 is discussed.

Styrene lower catabolic pathway in Pseudomonas fluorescens ST: identification and characterization of genes for phenylacetic acid degradation

Pseudomonas fluorescens ST is a styrene degrading microorganism that, by the sequential oxidation of the vinyl side chain, converts styrene to phenylacetic acid. The cluster of styrene upper pathway catabolic genes (sty genes) has been previously localized on a chromosomal region. This report describes the isolation, sequencing and analysis of a new chromosomal fragment deriving from the ST strain genomic bank that contains the styrene lower degradative pathway genes (paa genes), involved in the metabolism of phenylacetic acid. Analysis of the paa gene cluster led to the description of 14 putative genes: a gene encoding a phenylacetyl-CoA ligase (paaF), the enzyme required for the activation of phenylacetic acid; five ORFs encoding the subunits of a ring hydroxylation multienzymatic system (paaGHIJK); the gene paaW encoding a membrane protein of unknown function; five genes for a β-oxidation-like system (paaABCDE), involved in the steps following the aromatic ring cleavage; a gene encoding a putative permease (paaL) and a gene (paaN) probably involved in the aromatic ring cleavage. The function of some of the isolated genes has been proved by means of biotransformation experiments.

Characterization of Rhodococcus opacus R7, a strain able to degrade naphthalene and o-xylene isolated from a polycyclic aromatic hydrocarbon-contaminated soil

Rhodococcus opacus R7 was isolated from a soil contaminated with polycyclic aromatic hydrocarbons for its ability to grow on naphthalene. The strain was also able to degrade o-xylene, the isomer of xylenes most recalcitrant to microbial degradation. The catabolic pathways for naphthalene and o-xylene were investigated by identification of metabolites in R. opacus R7 cultures performed with the two hydrocarbons and by evaluation of some enzymes involved in the metabolism of these compounds. 1,2-Dihydro-1,2-dihydroxynaphthalene, salicylic and gentisic acids were identified as metabolites in cultures exposed to naphthalene. This suggests that the degradation occurs through the dioxygenation of the aromatic ring with the formation of 1,2-dihydro-1,2-dihydroxynaphthalene, dehydrogenated to the corresponding 1,2-dihydroxy derivative which is further oxidized to salicylic acid, a key intermediate of naphthalene metabolism; this compound is converted to gentisic acid cleaved by a gentisate 1,2-dioxygenase. From R. opacus R7 cultures supplied with o-xylene, 2,3-dimethylphenol and 3,4-dimethylcatechol were observed. The pathway of o-xylene involves the monooxygenation of the benzene nucleus leading to dimethylphenol which is further metabolised to 3,4-dimethylcatechol, followed by a meta cleavage reaction, catalyzed by the catechol 2,3-dioxygenase. R. opacus R7 is the first strain thus far described both in Gram-negative and Gram-positive bacteria which has the ability to degrade both a polycyclic aromatic hydrocarbon such as naphthalene and a monocyclic aromatic hydrocarbon such as o-xylene.

Cloning and comparison of mercury- and organomercurial-resistance determinants from a Pseudomonas stutzeri plasmid.

Plasmid pPB confers broad-spectrum mercury resistance (HgR) to a Pseudomonas stutzeri strain. Two pPB regions, separated by 25-30 kb and sharing homology with Tn501 mer (Hg detoxification) genes, were cloned separately and each was shown to carry a cluster of functional and independently regulated mer genes. One of the two gene clusters conferred resistance only to inorganic mercury, and had a structure identical to the classical model of narrow-spectrum mer operons. In the other cluster a novel merB gene, not homologous to the other known merB, but with the same function, was mapped upstream from merA, interposed between an organomercurial-responsive regulatory element and transport genes. Evidence suggests that merB and the other structural mer genes might be transcribed from two distinct promoters. The presence of two inverted repeat-like elements, identical to those of Tn5053, upstream from merR suggests that the pPB broad-spectrum-gene cluster could be part of a transposon-like element.

Biotransformation of styrenes by a Pseudomonas putida

SummaryA strain of Pseudomonas putida was isolated from soil in the presence of α-methylstyrene, as the sole carbon and energy source. The analysis of the oxidation products from culture broth allowed the identification of 2-phenyl-2-propen-1-ol and 1,2-dihydroxy-3-isopropenyl-3-cyclohexene suggesting the existence of different initial steps in the metabolism of α-methylstyrene. The same strain also oxidized styrene and produced by initial oxidation of the aromatic nucleus a compound identified as 1,2-dihydroxy-3-ethenyl-3-cyclohexene.

Bioconversion of substituted naphthalenes to the corresponding 1,2-dihydro-1,2-dihydroxy derivatives. Determination of the regio- and stereochemistry of the oxidation reactions

A mutant (TTC1) derived from Pseudomonas fluorescens N3 has been obtained for use in the bioconversion of several naphthalene derivatives to the corresponding optically active cis-dihydrodiols on a milligrams-to-grams scale. All main compounds have been characterized, their relative and absolute configuration assigned, and their enantiomeric purity determined. The regio- and stereoselectivity of the transformation has been established. The procedure therefore represents a valid method for the convenient preparation of a pool of valuable chiral syntons and auxiliaries.

Two aberrant mercury resistance transposons in the Pseudomonas stutzeri plasmid pPB.

The two mer operons of the Pseudomonas stutzeri OX plasmid pPB and their flanking regions have been sequenced and found to be part of two aberrant transposons. The narrow spectrum mer operon is almost identical to that of Tn501, but is associated with the remnants of Tn5053 tni genes rather than the Tn501 transposition module. The broad spectrum mer operon shows an overall homology with that of Tn5053, but differs from it in the presence of a merB gene, absent in Tn5053, and a merC gene instead of a merF. The pPB broad spectrum mer operon is associated with an incomplete Tn5053-like transposition module and with the Tn501 tnp genes, which are proximal, respectively, to the end and to the beginning of the mer operon. A hypothesis about pPB evolution is presented.

Alternative pathways foro-xylene orm-xylene andp-xylene degradation in aPseudomonas stutzeri strain

Pseudomonas stutzeri OX1 is able to grow ono-xylene but is unable to grow onm-xylene andp-xylene, which are partially metabolized through theo-xylene degradative pathway leading to the formation of dimethylphenols toxic to OX1.P. stutzeri spontaneous mutants able to grow onm-xylene andp-xylene have been isolated. These mutants soon lose the ability to grow ono-xylene. Data from HPLC analyses and from induction studies suggest that in these mutantsm-xylene andp-xylene could be metabolized through the oxidation of a methyl substituent.P. stutzeri chromosomal DNA is shown to share homology with pWW0 catabolic genes. In the mutant strains the region homologous to pWW0 upper pathway genes has undergone a genomic rearrangement.