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The rhizobacterium Pseudomonas aeruginosa M18 can produce a broad spectrum of secondary metabolites, including the antibiotics pyoluteorin (Plt) and phenazine-1-carboxylic acid (PCA), hydrogen cyanide, and the siderophores pyoverdine and pyochelin. The antibiotic biosynthesis of M18 is coordinately controlled by multiple distinct regulatory pathways, of which the GacS/GacA system activates Plt biosynthesis but strongly downregulates PCA biosynthesis. Here, we investigated the global influence of a gacA mutation on the M18 transcriptome and related metabolic and physiological processes. Transcriptome profiling revealed that the transcript levels of 839 genes, which account for approximately 15% of the annotated genes in the M18 genome, were significantly influenced by the gacA mutation during the early stationary growth phase of M18. Most secondary metabolic gene clusters, such as pvd, pch, plt, amb, and hcn, were activated by GacA. The GacA regulon also included genes encoding extracellular enzymes and cytochrome oxidases. Interestingly, the primary metabolism involved in the assimilation and metabolism of phosphorus, sulfur, and nitrogen sources was also notably regulated by GacA. Another important category of the GacA regulon was secretion systems, including H1, H2, and H3 (type VI secretion systems [T6SSs]), Hxc (T2SS), and Has and Apr (T1SSs), and CupE and Tad pili. More remarkably, GacA inhibited swimming, swarming, and twitching motilities. Taken together, the Gac-initiated global regulation, which was mostly mediated through multiple regulatory systems or factors, was mainly involved in secondary and primary metabolism, secretion systems, motility, etc., contributing to ecological or nutritional competence, ion homeostasis, and biocontrol in M18. [ABSTRACT FROM AUTHOR]

Phenazine-1-carboxylic acid (PCA) production was enhanced in Pseudomonas sp. M18 wild strain and its mutants carrying recombinant pME6032Phz for phz gene cluster overexpression, among which Pseudomonas sp. strain M18GQ/pME6032Phz, a gacA and qscR double gene chromosomally inactivated mutant harboring pME6032Phz, showed the highest PCA yield. The conditions for fermentation and isopropyl-β- d-1-thiogalactopyranoside (IPTG) induction were optimized for strain M18GQ/pME6032Phz in shake flask experiments. A one-factor-at-a-time approach, followed by a fractional factorial design identified soybean meal, corn steep liquor, and ethanol as statistically significant factors. Optimal concentrations and mutual interactions of the factors were then determined by the method of steepest ascent and by response surface methodology based on the center composite design. The predicted PCA production was 6,335.2 mg/l after 60 h fermentation in the optimal medium of 65.02 g soybean meal, 15.36 g corn steep liquor, 12 g glucose, 21.70 ml ethanol, and 1 g MgSO4 per liter in the flask fermentations, with induction of 1.0 mmol/l IPTG 24 h after inoculation. In an experimental validation under these conditions, the maximum PCA production was 6,365.0 mg/l. This represents a ∼60% increase over production by strain M18GQ in optimal conditions. The negative effect of plasmid pME6032 on the expression of chromosomally located phz gene cluster was found in Pseudomonas sp. M18GQ, and the possible reason was discussed in the text. [ABSTRACT FROM AUTHOR]

The qscR gene, encoding a quorum sensing regulator, was cloned and the qscR-null mutant strain M-18Q derived from Pseudomonas sp. M-18 was constructed to study the effect of the qscR gene on biosynthesis of phenazine-1-carboxylic acid (PCA) and pyoluteorin (Plt) in strain M-18. Results showed that the PCA produced in the mutant increased four- to six-fold, while the synthesis of Plt was barely influenced in comparison with the wild type. The results were confirmed by complementation with the qscR gene in trans in strain M-18Q. The negative effect of the qscR gene on PCA production was further confirmed by analysis of β-galactosidase activities from the translational phzA’-lacZ’ fusion. Furthermore, by introducing a qscR-lacZ transcriptional fusion vector to strains M-18, M-18Q, and M-18G, a gacA inactivation mutant in strain M-18, respectively, it was found that β-galactosidase activity in both strain M-18G and strain M-18Q was decreased to half that in the wild type. This suggested that QscR might be involved in autoinducing its own gene expression and act as an intermediate in GacA-dependent gene regulation as well. The result was further demonstrated by the overexpression of the gacA gene in strain M-18Q. [ABSTRACT FROM AUTHOR]

In this paper, the sensitivity of Escherichia coli to surfactin and fengycin was observed, and the optimization of the antimicrobial activity of surfactin and fengycin to E. coli in milk by a response surface methodology was studied. Results showed that E. coli had high sensitivity to these antibiotics, whose minimal inhibitory concentrations were 15.625 μg·mL−1 and 31.25 μg·mL−1, respectively. The optimization result indicated that E. coli could be sterilized by 5 orders of magnitude when the temperature was 5.5°C, the action time was 15.8 h, and the concentration (surfactin/fengycin weight ratio 1:1) was 14.63 μg·mL−1. [ABSTRACT FROM AUTHOR]

BACKGROUND: Trichloroethylene (TCE) is used extensively as an industrial solvent and has been recognized as one of the major environmental pollutants. To date, > 200 cases of TCE-induced hypersensitivity dermatitis among exposed workers have been reported worldwide, and TCE exposure has become one of the critical occupational health issues in Asia. OBJECTIVES: The study aimed to identify genetic susceptible biomarkers associated with the TCE- induced hypersensitivity dermatitis in genes located in the human leukocyte antigen (HLA) region. METHODS: From 1998 to 2006, 121 cases with TCE-induced hypersensitivity dermatitis and 142 tolerant controls were recruited into the population-based case-control study. We determined HLA alleles B, DRB1, DQA1, and DQB1, by sequence-based typing. p-Values were corrected for comparisons of multiple HLA alleles. In addition, we compared and analyzed the structure character of amino acid residues of HLA molecules found in participants. RESULTS: We obtained complete genotyping data of 113 cases and 142 controls. The allele HL.4-B*1301 was present in 83 (73.5%) of 113 patients compared with 13 (9.2%) of 142 tolerant workers (odds ratio = 27.5; 95% confidence interval, 13.5-55.7; corrected p = 1.48 × 10-21). In addition, the HLA-B*44 alleles were present in 6.2% (7/113) of patients, but were absent in TCE- tolerant workers. Residue 95 shared by HLA-B*1301 and HLA-B*44 molecules formed a different pocket F than other residues. CONCLUSIONS: The allele HLA-B*1301 is strongly associated with TCE-induced hypersensitivity dermatitis among exposed workers and might be used as a biomarker to predict high risk individuals to TCE. [ABSTRACT FROM AUTHOR]

A new Pseudomonas strain, designated GP72, was isolated from green pepper rhizosphere and identified as a member of species Pseudomonas chlororaphis based on morphology; conventional biochemical and physiologic tests; Biolog GN system (Biolog Inc., Hayward, CA); and 16S rDNA sequence analysis. The secondary metabolites produced by this strain have shown broad-spectrum antifungal activity against various phytopathogens of agricultural importance in vitro. Two main antifungal substances produced by this strain proved to be phenazine-1-carboxylic acid and 2-hydroxyphenazine with further purification and structure elucidation based on ultraviolet-absorbent spectrum scanning, atmospheric pressure chemical ionization–mass spectrometry (APCI-MS) spectrum, and 1H,13C nuclear magnetic resonance spectrums. Strain GP72 could produce quorum-sensing signaling molecules of N-butanoyl-L-homoserine lactone and N-hexanoyl-L-homoserine lactone, which were found to accumulate with different quantities in King’s medium B and pigment producing medium, respectively. [ABSTRACT FROM AUTHOR]

Bacillus subtilis fmbj can produce a lipopeptide antimicrobial substance, the main components of which are surfactin and fengycin. In this paper, the sensitivity of Bacillus cereus to antimicrobial lipopeptides from B. subtilis fmbj was observed, and the effect of the microstructure of antimicrobial lipopeptide on spores of B. cereus was investigated. At the same time, the optimization of the inactivation of antimicrobial lipopeptides to spores of B. cereus by a response surface methodology was studied. Results showed that B. cereus had high sensitivity to it, whose minimal inhibitory concentration was 156.25 μg/ml. It could result in the death of spores by destroying the structure of resting spores and sprouting spores, as was observed by transmission electron microscopy. The optimization result indicated that spores of B. cereus could be inactivated by 2 orders of magnitude when the temperature was 29.6°C, the action time was 7.6 h, and the concentration was 3.46 mg·ml−1. [ABSTRACT FROM AUTHOR]

Plant growth promoting rhizobacteria (PGPR) strain Pseudomonas sp. M18 can produce two different types of antibiotics, pyoluteorin (Plt) and phenazine-1-carboxylic acid (PCA). The global regulator RsmA is a translational repressor of secondary metabolism in many prokaryotes. A chromosomally rsmA inactivated mutant strain M18R was constructed to study the regulatory mechanism of Plt and PCA biosynthesis and enhancement of Plt or PCA production in Pseudomonas sp. M18. The accumulation of Plt increased six-fold over that of the wild-type strain whereas PCA production was not significantly affected in cultures of M18R. Plt production was inhibited completely but PCA biosynthesis was not altered after complementation with rsmA gene in trans in the strain of M18R. The differential activity of rsmA gene on these two operons was further confirmed by the analysis of β-galactosidase activities from translational phzA′-′lacZ and pltA′-′lacZ fusion, in which phzA is the first enzyme gene of the phenazine biosynthesis pathway and pltA is the first gene of the pyoluteorin biosynthesis pathway. The results indicate that RsmA can control Plt production negatively but not PCA production in M18, and show that the global regulator RsmA does not repress the biosynthesis of all secondary metabolites. [ABSTRACT FROM AUTHOR]

BACKGROUND RESULTS CONCLUSION Sea buckthorn (Hippophae rhamnoides L.) pulp oil is rich in functional components; however, low water solubility and stability limit its applications. This study fabricated sea buckthorn pulp oil microcapsules using whey protein isolate (WPI), soy protein isolate (SPI), sodium caseinate (NaCN), gum arabic (GA), starch sodium octenylsuccinate (OSAS) and SPI mixed with chitosan (CHI). The influences of these wall materials on physicochemical properties, release behavior and digestibility were explored.Protein‐based wall materials (WPI, NaCN, SPI) demonstrated lower bulk densities due to their porous structures and larger particle sizes, while GA and OSAS produced denser microcapsules. Encapsulation efficiency was the highest for protein‐based microcapsules (79.41–89.12%) and the lowest for GA and OSAS. The surface oil percentage of protein‐based microcapsules (1.41–4.40%) was lower than that of the other microcapsules. Protein‐based microcapsules showed concave and cracked surfaces, while GA and OSAS microcapsules were spherical and smooth. CHI improved reconstitution performance, leading to faster dissolution. During simulated gastrointestinal digestion, protein‐based microcapsules released more free fatty acids (FFAs) in the intestinal phase, while CHI‐modified SPI microcapsules showed a delayed release pattern due to thicker walls.Protein‐based wall materials were more effective for sea buckthorn pulp oil microencapsulation, providing higher encapsulation efficiency, better flow properties and releasing more FFAs. The addition of CHI led to the layer‐by‐layer self‐assembly of the microcapsule wall and resulted in sustained release during in vitro intestinal digestion. These findings suggested the potential of protein‐based microcapsules for targeted delivery and improved applications of bioactive oils in the food industry. © 2024 Society of Chemical Industry. Published by John Wiley & Sons Ltd [ABSTRACT FROM AUTHOR]