The marine bacteria Shewanella frigidimarina NCIMB400 upregulates the type VI secretion system during early biofilm formation.

Shewanella sp. are facultative anaerobic Gram-negative bacteria, extensively studied for their electron transfer ability. Shewanella frigidimarina has been detected and isolated from marine environments, and in particular, from biofilms. However, its ability to adhere to surfaces and form a biofilm is poorly understood. In this study, we show that the ability to adhere and to form a biofilm of S. frigidimarina NCIMB400 is significantly higher than that of Shewanella oneidensis in our conditions. We also show that this strain forms a biofilm in artificial seawater, whereas in Luria-Bertani, this capacity is reduced. To identify proteins involved in early biofilm formation, a proteomic analysis of sessile versus planktonic membrane-enriched fractions allowed the identification of several components of the same type VI secretion system gene cluster: putative Hcp1 and ImpB proteins as well as a forkhead-associated domain-containing protein. The upregulation of Hcp1 a marker of active translocation has been confirmed using quantitative reverse transcription polymerase chain reaction. Our data demonstrated the presence of a single and complete type VI secretion system in S. frigidimarina NCIMB400 genome, upregulated in sessile compared with planktonic conditions. The fact that three proteins including the secreted protein Hcp1 have been identified may suggest that this type VI secretion system is functional.

Catabolism of L-ascorbate by Lactobacillus rhamnosus GG.

Catabolism of L-ascorbate by enteric bacteria is well documented, but no study has formally proved that bacteria of the Lactobacillus genus ferment this compound. However, some genes analogous to those of yiaK-S operon and ula regulon, which encoded proteins leading to L-ascorbate degradation by Escherichia coli and Klebsiella pneumoniae , have been identified in the recently sequenced Lactobacillus rhamnosus GG genome. Investigations by HPLC and in vivo (13)C NMR using L-[1,6-(13)C]-ascorbate showed that L. rhamnosus GG, a common probiotic strain, has the ability to catabolize L-ascorbate under anaerobiosis. The main products of the ascorbate degradation have been identified as CO(2), acetate, and lactate. These results are in accordance with the metabolic pathway proposed for the fermentation of L-ascorbate by E. coli.

Production of trans-2-methyl-5-isopropylhexa-2,5-dienoic acid by Pseudomonas rhodesiae CIP 107491.

The feasibility of trans-2-methyl-5-isopropylhexa-2,5-dienoic acid (novalic acid) accumulation using the alpha-pinene degradation pathway of Pseudomonas rhodesiae CIP 107491 was studied. This appeared possible by using concentrated living bacterial cells produced under oxygen limitation with alpha-pinene as sole carbon source. The second step of the process, the bioconversion itself, had to be performed without oxygen limitation due to the need for cofactor regeneration. Results showed that a not yet reported cofactor-dependent enzymatic isomerization of isonovalal into novalal was likely to occur and that both aldehyde isomers could be oxidized to the corresponding acid. Precursors tested, alpha-pinene oxide and isonovalal had a strong permeabilization effect on bacterial cells. This effect, which increased from the oxide to the aldehyde, led to an inactivation of the respiratory chain and to acids synthesis stop. Present results allowed to obtain about 12 g/L acids (80% novalic acid) with an average yield close to 50% after 12h reaction in a biphasic system using alpha-pinene oxide as precursor .