Effect of pps disruption and constitutive expression of srfA on surfactin productivity, spreading and antagonistic properties of Bacillus subtilis 168 derivatives.

AIMS: To analyse the effects of plipastatin operon disruption and constitutive expression of surfactin operon in Bacillus subtilis 168 on surfactin productivity, in vitro invasive growth and antagonism against fungi. METHODS AND RESULTS: The srfA native promoter was replaced by the constitutive promoter P(repU) in B. subtilis 168 after integration of a functional sfp gene. Moreover, the plipastatin synthesis was further disrupted in the B. subtilis 168 derivatives. In liquid media, an earlier and higher expression of P(repU), than that found with P(srfA), led to a specific surfactin production fivefold higher after 6 h of culture. On solid media, not only the invasive growth and the haemolytic activity but also the antifungal activity of the constitutive strains were improved when compared to the parental strain BBG111. As expected, the disruption of the plipastatin operon strongly reduced in vitro antifungal properties but, interestingly, enhanced specific surfactin production (1.47 g g(-1) of biomass), spreading behaviour and haemolytic activity of the strains. CONCLUSIONS: This work demonstrates for the first time the interdependency of surfactin and plipastatin regarding their biosynthesis as well as their influence on the biological activities of the producing strain. SIGNIFICANCE AND IMPACT OF THE STUDY: The constitutive overproduction of surfactin enhances the invasive growth and the in vitro antagonistic activity of the mutant strain. Both properties are known to play an important role in the biocontrol of plant diseases. Plipastatin operon disruption increases the surfactin productivity of mutant strains. These mutants are interesting for use in continuous bioprocesses for surfactin production or in bioremediation.

The mineralization of 5-amino-2,4,6-triiodoisophthalic acid by a two-stage fixed-bed reactor.

Iodinated X-ray contrast media have been detected in hospital effluent, sewage treatment plant effluent, rivers and groundwater aquifers. No process has been developed to remove triiodinated aromatic molecules. In this paper, we present a biological sequential process using an anaerobic fixed-bed reactor coupled in series with an aerobic fixed-bed reactor for degrading 5-amino-2,4,6-triiodoisophthalic acid (ATIA), the core structure of a X-ray contrast media family. The results obtained showed that the coupled reactor eliminated up to 870+/-44 mg of carbon L(-1) day(-1), with a molar ethanol/ATIA ratio of 4 in the feeding medium. The anaerobic reactor (ANR) undertook the majority of the deiodination of the aromatic nucleus and had a maximum deiodination rate of 23.4+/-0.06 mM day(-1). The aerobic reactor (AER) mineralized ATIA and was also able to eliminate its metabolites. This study suggests that the mineralization of ATIA can be achieved efficiently in a coupled anaerobic-aerobic bioreactor.

Adsorption of surfactin produced from Bacillus subtilis using nonwoven PET (polyethylene terephthalate) fibrous membranes functionalized with chitosan.

This article deals with an alternative method for bio-separation of surfactin produced by Bacillus subtilis using sorption method on nonwoven PET (polyethylene terephthalate) fibrous membranes functionalized with chitosan. In the first part of the study, surface functionalization of the PET nonwoven fibrous membranes is carried out with aqueous 65% deacetylated chitosan solution with or without a prior surface activation using air-atmospheric plasma treatment. Very small modification of the PET fibrous nonwoven air-permeability confirms the functionalization of PET fibre surface with little reduction of membrane porosity. The functionalized membranes are then characterized by physico-chemical methods: X-ray Photoelectron Spectroscopy (XPS), Wettability and zeta potential. Chitosan increases drastically the zeta potential of PET at all pH values though a prior plasma treatment of the PET membrane reduces slightly the increase in zeta potential values. Sorption of surfactin quantified by HPLC shows that the extent of surfactin sorption on PET nonwovens depends on the surface functionalization method. Surface functionalization with chitosan results in immediate sorption of the entire quantity of surfactin. A prior surface activation by air atmospheric plasma treatment of the PET membranes before chitosan application retards the sorption of entire surfactin which takes place after 1.5h, only. Increased zeta potential and increased hydrophobic behavior in the presence of chitosan without plasma activation would explain the interesting surfactin sorption results.

Adhesion of a Pseudomonas putida strain isolated from a paper machine to cellulose fibres.

The adhesion to cellulose fibres of a strain of Pseudomonas putida isolated from a paper machine was studied under different environmental conditions. The physicochemical properties of both P. putida cells and cellulose fibres were also determined to better understand the adhesion phenomenon. Adhesion was rapid (1 min) and increased with time, cell concentration and temperature (from 25 to 40 degrees C), indicating that bacterial adhesion to cellulose fibres is essentially governed by a physicochemical process. The P. putida cell surface was negatively charged, as shown by electrophoretic mobility measurements, and was hydrophilic due to a strong electron-donor character, as shown by the microbial adhesion to solvents method. Cellulose fibres were shown to be hydrophilic by contact angle measurements using the capillary rise method. These results suggest the importance of Lewis acid-base interactions in the adhesion process. In various ionic solutions (NaCl, KCl, CaCl(2) and MgCl(2)), adhesion increased with increasing ionic strength up to 10-100 mM, indicating that, at low ionic strength, electrostatic interactions were involved in the adhesion process. An increase in the C/N ratio of the growth medium (from 5 to 90) decreased adhesion but this could not be related to changes in physicochemical properties, suggesting that other factors may be involved. In practice, temperature, ionic strength and nitrogen concentration must be taken into consideration to reduce bacterial contamination in the paper industry.

Enrichment and properties of an anaerobic mixed culture that reductively deiodinates 5-amino-2,4,6-triiodoisophthalic acid, an X-ray contrast agent precursor.

5-Amino-2,4,6-triiodoisophthalic acid (ATIA), both a precursor and a degradative intermediate of triiodinated contrast media, was anaerobically converted by sludge from a wastewater treatment plant. ATIA conversion took place only when an electron donor such as ethanol was added. A stable mixed culture was established by transfer to a defined synthetic mineral medium with ATIA and ethanol. It could be maintained for 1 year when the sulfate concentration was kept below 30 microM. Transient appearance of 5-amino-2,4-diiodoisophthalic acid, iodide release (2.7 mol iodide/mol ATIA) and accumulation of 5-aminoisophthalic acid indicated that ATIA was reductively dehalogenated. The enriched mixed culture also dehalogenated ATIA derivatives but deiodination remained incomplete. ATIA was the sole terminal electron acceptor used by the mixed culture during deiodination. The ratio of electrons transferred to ATIA, 0.83, was consistent with a respiratory metabolism. Formate, acetate, lactate, butyrate and hydrogen were also used as electron donors. Deiodination was inhibited by a headspace of air or by addition of nitrate, sulfite or thiosulfate. The reaction was 2.6 times slower with sulfate than without.

Phylogenetic analysis of an anaerobic microbial consortium deiodinating 5-amino-2,4,6-triiodoisophthalic acid.

The dehalogenating performance of an anaerobic 5-amino-2,4,6-triiodoisophthalic acid (ATIA) fixed-bed reactor was evaluated. The reactor operating conditions were set for ATIA deiodination. A phylogenetic survey for a stable anaerobic ATIA-deiodinating microbial consortium was carried out using 16S rDNA restriction fragment length polymorphism, and unique clones were sequenced. Four phylotypes were identified. Two sequences were related to those of Desulfitobacterium frappieri species and another was closest to that of Desulfitobacterium hafniense, but may have represented a new Desulfitobacterium species. Desulfitobacteria were previously described as aryl-dechlorinating and debrominating bacteria. The new strains identified in this study were probably responsible for the ATIA deiodination. The fourth clone was related to the Clostridium-Flavobacterium-Bacteroides group.