Genes encoding L-sorbose metabolism of Lactobacillus casei ATCC 393 have been identified on a 6.8-kb chromosomal DNA fragment. Sequence analysis revealed seven complete genes and a partial open reading frame transcribed as two units. The deduced amino acid sequences of the first transcriptional unit (sorRE) showed high similarity to the transcriptional regulator and the L-sorbose-1-phosphate reductase of the sorbose (sor) operon from Klebsiella pneumoniae. The other genes are transcribed as one unit (sorFABCDG) in opposite direction to sorRE. The deduced peptide sequence of sorF showed homology with the D-sorbitol-6-phosphate dehydrogenase encoded in the sor operon from K. pneumoniae and sorABCD to components of the mannose phosphotransferase system (PTS) family but especially to domains EIIA, EIIB, EIIC and EIID of the phosphoenolpyruvate-dependent L-sorbose PTS from K. pneumoniae. Finally, the deduced amino acid sequence of a truncated gene (sorG) located downstream of sorD presented high similarity with ketose-1,6-bisphosphate aldolases. Results of studies on enzyme activities and transcriptional analysis revealed that the two gene clusters, sorRE and sorFABCDG, are induced by L-sorbose and subject to catabolite repression by D-glucose. Data indicating that the catabolite repression is mediated by components of the PTS elements and by CcpA, are presented. Results of sugar uptake assays in L. casei wild-type and sorBC mutant strains indicated that L-sorbose is taken up by L-sorbose-specific enzyme II and that L. casei contains an inducible D-fructose-specific PTS. Results of growth analysis of those strains and a man sorBC double mutant suggested that L-sorbose is probably also transported by the Dmannose PTS. We also present evidence, from studies on a sorR mutant, suggesting that the sorR gene encodes a positive regulator of the two sor operons. Sequence alignment of SorR, SorC (K. pneumoniae), and DeoR (Bacillus subtilis) revealed that they might constitute a new group of transcriptional regulators.In the enteric bacteria Klebsiella pneumoniae and Escherichia coli, the ketose L-sorbose is transported and phosphorylated through a phosphoenolpyruvate-dependent L-sorbose-specific phosphotransferase system (PTS) (36,46,47). The metabolism of L-sorbose involves the formation of D-fructose-6-phosphate, which enters the glycolysis pathway (Fig. 1A). During uptake of a PTS carbohydrate, the common PTS proteins, enzyme I (EI) and HPr, transfer a phosphoryl group from phosphoenolpyruvate to the substrate-specific EII complexes. The L-sorbose PTS EII consists of two membrane-bound proteins, EIIC Sor PTS elements are involved in sugar uptake and also in the regulation of chemotaxis, carbon metabolism, and modulation of gene expression (9,20,27). Many catabolic operons in bacteria are subject to carbon catabolite repression (CR) by rapidly metabolizable carbon sources, especially glucose (31, 37). The main CR mechanism identified in E. coli can be summarized as follows. EIIAB Glc acts as the sensor of extracellular gl...

A 2-deoxy-~-glucose-resistant mutant of a pLZl5-cured derivative of Lactobadhs casei ATCC 393 was isolated on agar medium containing 10 mM 2-deoxy-~-glucose and 5 g lactose I-'. The mutant was impaired in the main glucose transport mechanism, a PTSman-type system. Additionally a protonmotive-force-dependent glucose permease was detected. The growth response and the sugar consumption rates of the wild-type and the PTSmn-deficient mutant suggested that the mutated element of the complex IIABCman was, in the wild-type, responsible for a strong repression by glucose and mannose of the lactose and ribose assimilation genes, while assimilation of galactose was only weakly repressed. It is postulated that they are regulated by a different mechanism of catabolite repression.

Elsevier Cajuste, JF.; Gonzalez-Candelas, L.; Veyrat, A.; García-Breijo, F.; Reig Armiñana, J.; Lafuente, MT. (2010) The effect of ethylene (2 L L −1 ) on total and soft epicuticular wax content and wax morphology has been investigated in mature 'Navelate' (Citrus sinensis, L. Osbeck) oranges held under non-stressful environmental conditions (22• C and constant high relative humidity (90-95% RH)). In addition, the objective of the study was to understand whether the ethylene-induced changes in epicuticular wax might participate in the beneficial effect of ethylene reducing non-chilling peel pitting, by modifying peel water, osmotic or turgor potential, or disease incidence caused by Penicillium digitatum (Pers.:Fr.) Sacc. Ethylene increased total and soft epicuticular wax content in 'Navelate' fruit and induced structural changes in surface wax that might be related to the formation of new waxes. Changes in epicuticular wax morphology, but not in its content, might be involved in the protective role of ethylene reducing non-chilling peel pitting, although the beneficial effect of the hormone is not related to water stress. Cell water and turgor potentials in freshly harvested fruit and fruit stored in air under non-stressful conditions suggest that water stress is not a limiting factor leading to the development of this physiological disorder. In addition, the results indicated that formation of new waxes in fruit treated with ethylene may partially cover stomata, cracks or areas lacking wax occurring in stored fruit and is likely to improve physical barriers to P. digitatum penetration.

In this study, we evaluated the activity of short antimicrobial peptides against different fungal isolates that cause postharvest decay of fresh fruits. The previously identified hexapeptides PAF19, PAF26 and LfcinB4-9 inhibited the in vitro growth of isolates from Penicillium digitatum and P. italicum, and from Alternaria and Geotrichum genera, being no active against Rhizopus, Mucor and Aspergillus. The results extend our previous observations on the specific and distinct activity profiles of this class of antifungal peptides. In addition, peptide activities were compared with that of two fungicides used for citrus fruit preservation, thiabendazole (TBZ) and imazalil (IMZ). We observed a lack of correlation between peptide and fungicide sensitivity among different species. Importantly, P. digitatum and P. italicum isolates resistant to fungicides were susceptible to peptides and our data suggest that common multiple drug resistance mechanisms are not active against this class of peptides. The in vitro peptide inhibition was correlated with a retard of the decay caused by Penicillium on citrus fruits, and this effect was comparable for both fungicide-resistant and -sensitive isolates. Comparison of PAF26 and TBZ in vitro minimum inhibitory concentration (MIC) values and their in vivo effect on citrus decay indicated that PAF26 performed in vivo better than TBZ.