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This study demonstrates the effect of fermented botanical product (FBP) on Ralstonia pseudosolanacearum -induced bacterial wilt disease and unravels its action mechanism. Soaking with diluted FBP solutions (0.1%-0.5%) significantly suppressed bacterial wilt in tomato plants, and FBP-treated tomato plants grew well against R. pseudosolanacearum infection. Growth assays showed that FBP had no antibacterial effect but promoted R. pseudosolanacearum growth. In contrast, few or no R. pseudosolanacearum cells were detected in aerial parts of tomato plants grown in FBP-soaked soil. Subsequent infection assays using the chemotaxis-deficient mutant (Δ cheA) or the root-dip inoculation method revealed that FBP does not affect pathogen migration to plant roots during infection. Moreover, FBP-pretreated tomato plants exhibited reduced bacterial wilt in the absence of FBP. These findings suggest that the plant, but not the pathogen, could be affected by FBP, resulting in an induced resistance against R. pseudosolanacearum , leading to a suppressive effect on bacterial wilt. [ABSTRACT FROM AUTHOR]

3-Hydroxypropionic acid (3-HP) and 1,3-propanediol (1,3-PDO) have tremendous potential markets in many industries. This study evaluated the simultaneous biosynthesis of the 2 compounds using the new psychrophile-based simple biocatalyst (PSCat) reaction system. The PSCat method is based on the expression of glycerol dehydratase, 1,3-propanediol dehydrogenase, and aldehyde dehydrogenase from Klebsiella pneumoniae in Shewanella livingstonensis Ac10 and Shewanella frigidimarina DSM 12253, individually. Heat treatment at 45 °C for 15 min deactivated the intracellular metabolic flux, and the production process was started after adding substrate, cofactor, and coenzyme. In the solo production process after 1 h, the maximum production of 3-HP was 62.0 m  m. For 1,3-PDO, the maximum production was 25.0 m  m. In the simultaneous production process, productivity was boosted, and the production of 3-HP and 1,3-PDO increased by 13.5 and 4.9 m  m , respectively. Hence, the feasibility of the individual production and the simultaneous biosynthesis system were verified in the new PSCat approach. [ABSTRACT FROM AUTHOR]

We have demonstrated that chemotaxis to l -malate facilitated motility of Ralstonia pseudosolanacearum MAFF 106611, a causative agent of bacterial wilt, to plant roots. Here, we evaluated the assumption that the disruption of chemotaxis to l -malate leads to inhibition of plant infection by R. pseudosolanacearum MAFF 106611. Chemotactic assays revealed that chemotaxis to l -malate was completely or partially inhibited in the presence of l -, d -, and dl -malate, respectively. Moreover, l -malate served as a carbon and energy source for R. pseudosolanacearum MAFF 106611, while d -malate inhibited the growth of this bacterium. In the sand-soak inoculation virulence assay for tomato plants, the addition of l -, d -, and dl -malate to sand suppressed the plant infection. We concluded that supplementation of l - and dl -malate suppresses tomato plant infection with R. pseudosolanacearum MAFF 106611 by disrupting its chemotaxis to l -malate, while d -malate suppresses it by both the disruption of l -malate chemotaxis and inhibition of growth. [ABSTRACT FROM AUTHOR]

Pseudomonas protegens CHA0, known as plant-growth-promoting rhizobacterium, showed positive chemotactic responses toward proteinaceous L-amino acids. Genomic analysis revealed that P. protegens CHA0 possesses four putative chemoreceptors for amino acids (designated CtaA, CtaB, CtaC, and CtaD, respectively). Pseudomonas aeruginosa PCT2, a mutant defective in chemotaxis to amino acids, harboring a plasmid containing each of ctaA, ctaB, ctaC, and ctaD showed chemotactic responses to 20, 4, 4, and 11 types of amino acids, respectively. To enhance chemotaxis toward amino acids, we introduced the plasmids containing ctaA, ctaB, ctaC, or ctaD into P. protegens CHA0. By overexpression of the genes, we succeeded in enhancing chemotaxis toward more than half of the tested ligands. However, unexpectedly, the P. protegens CHA0 transformants showed unchanged or decreased responses to some amino acids when compared to wild-type CHA0. We speculate that alternation of expression of a chemoreceptor may affect the abundance of other chemoreceptors. cDNA: complementary DNA; LBD: ligand-binding domain; MCP: methyl-accepting chemotaxis protein; PDC: PhoQ/DcuS/CitA; PGPR: plant-growth-promoting rhizobacteria; qRT-PCR: quantitative reverse transcription PCR Four chemotaxis sensors for amino acids in PGPRPseudomonas protegens CHA0. Amino acid chemotaxis can be enhanced by overexpression of the genes. [ABSTRACT FROM AUTHOR]

We previously constructed a Psychrophile-based Simple bioCatalyst (PSCat) reaction system, in which psychrophilic metabolic enzymes are inactivated by heat treatment, and used it here to study the conversion of aspartic acid from fumaric acid mediated by the activity of aspartate ammonia-lyase (aspartase). In Escherichia coli, the biosynthesis of aspartic acid competes with that of L-malic acid produced from fumaric acid by fumarase. In this study, E. coli aspartase was expressed in psychrophilic Shewanella livingstonensis Ac10 heat treated at 50 °C for 15 min. The resultant PSCat could convert fumaric acid to aspartic acid without the formation of L-malic acid because of heat inactivation of psychrophilic fumarase activity. Furthermore, alginate-immobilized PSCat produced high yields of aspartic acid and could be re-used nine times. The results of our study suggest that PSCat can be applied in biotechnological production as a new approach to increase the yield of target compounds. [ABSTRACT FROM AUTHOR]

A transformation system for Moorella thermoacetica ATCC39073 was developed using thermostable kanamycin resistant gene ( kanR) derived from the plasmid pJH1 that Streptococcus faecalis harbored. When kanR with its native promoter was introduced into uracil auxotrophic mutant of M. thermoacetica ATCC39073 together with a gene to complement the uracil auxotrophy as a selection marker, it did not give kanamycin resistance due to poor transcription level of kanR. However, the use of glyceraldehyde-3-phosphate dehydrogenase promoter cloned from M. thermoacetica ATCC39073 significantly improved transcription level of kanR and resulted in the cell growth in the presence of more than 150 μg mL−1 kanamycin. It was also demonstrated that kanR with G3PD promoter can be used as a selection marker for transformation of wild-type strain of M. thermoacetica ATCC39073. [ABSTRACT FROM AUTHOR]

Aniline and chlorinated anilines (CAs) are classified as priority pollutants; therefore, an effective method for detection and monitoring is required. In this study, a green-fluorescence protein-based bioreporter for the detection of aniline and CAs was constructed in Escherichia coli DH5α, characterized and tested with soil and wastewater. The sensing capability relied on the regulatory control between a two-component regulatory protein, TodS/TodT, and the P promoter of Pseudomonas putida T-57 (PpT57), since the gene expression of todS, todT, and todC2 are positively induced with 4-chloroaniline. The bioreporter system (DH5α/pPXGFP-pTODST) is markedly unique with the two co-existing plasmids. The inducibility of the fluorescence response was culture-medium- and time-dependent. Cells grown in M9G medium exhibited a low background fluorescence level and were readily induced by 4CA after 3-h exposure, reaching the maximum induction level at 9 h. When tested with benzene, toluene, ethyl-benzene and xylene, aniline and CAs, the response data were best fit by a sigmoidal dose-response relationship, from which the K value was determined for the positive effectors. 3CA and 4CA were relatively powerful inducers, while some poly-chlorinated anilines could also induce green fluorescence protein expression. The results indicated a broader recognition range of PpT57'sTodST than previously reported for P. putida. The test results with environmental samples were reliable, indicating the potential application of this bioreporter in the ecotoxicology assessment and bioremediation of areas contaminated with aniline- and/or CAs. [ABSTRACT FROM AUTHOR]

The article focuses on a study related to the identification of the replication region of circular plasmid from Rhodococcus opacus. As per the study, a Polymerase Chain Reaction (PCR)-based deletion analysis is performed followed by restriction digestion and PCR-amplification analyses. The study further concludes that product of Open Reading Frame (ORF) and its orthologs serve as novel replication proteins for circular bacterial plasmids.

Brevibacillus agri strain 13 was isolated and characterized as a Gram-positive organic-solvent-tolerant bacterium able to grow at 45 °C. It can tolerate high concentrations (5% and 20%, v/v) of various organic solvents with a broad range of log Pow when the organic solvent was provided as a nonaqueous layer. Although it can tolerate a number of aromatic solvents, it cannot utilize them as a sole carbon source. The surface characteristics of cells exposed to organic solvent were investigated using the bacterial adhesion to hydrocarbon test, a contact angle measurement, ζ potential determination, and fluorescence microscopy analysis and compared with that of nonexposed cells. The results showed that although it has a hydrophilic cell surface, it has a unique indigenous cell surface characteristic in which the cells can stabilize solvent-in-water emulsion by adhering to the solvent–water interface of the solvent droplets. The tolerance and predilection of B. agri strain 13 toward organic solvents may suggest its potential application as a whole-cell biocatalyst for the biotransformation process of water-immiscible substrate(s). [ABSTRACT FROM AUTHOR]

A solvent-tolerant, slightly thermophilic bacterium was isolated at 45 °C in the presence of toluene vapor provided as the sole carbon source. Strain T27 was identified as Deinococcus geothermalis T27. It could tolerate high concentrations of solvent provided as a nonaqueous layer (5% and 20%, v/v) to a cell suspension and had a remarkable ability to tolerate a broad range of solvents having log Pow values ranging from 5.6 of n-decane to as low as 0.7 of ethyl acetate. It was also able to utilize some of the solvents tested as a growth substrate at 45 °C. The addition of Ca2+ ion, glucose and fructose partially promoted solvent tolerance. Cells exposed to ethyl acetate appeared to have a smaller size; however, the cell structure was not altered and was apparently well defined even after solvent shock. The tolerance of D. geothermalis T27 in the presence of high levels of toxic solvent stress at a comparatively high temperature indicated its potential use in biotechnological applications as well as bioremediation of xenobiotics. [ABSTRACT FROM AUTHOR]

We examined performance of mice in discrimination of liquor odors by Y-maze behavioral assays. Thirsty mice were initially trained to choose the odor of a red wine in the Y-maze. After successful training (>70% concordance for each trained mouse), the individual mice were able to discriminate the learned red wine from other liquors, including white wine, rosé wine, sake, and plum liqueur. However, when the mice were tested to distinguish fine differences between 2 brands of red wine, their performance significantly varied among the individual trained mice. Among 10 mice tested, 2 mice were able to discriminate between the red wines (>75% concordance) whereas 6 mice failed to distinguish between them (50–67% concordance, where chance could be assumed to be 50%). More importantly, 2 other mice exhibited lower than 30% concordance, indicating that they were more attracted to the nonrewarded red wine compared with the learned one. This result suggested that the individual mice directed attention to different subsets of volatile components emanating from the rewarded red wine, when they were trained to choose the liquor odor in the Y-maze. Selective attention of mice was also observed in Y-maze behavioral assays using the mixtures of 3 or less pure odorants. Additionally, we also observed that the olfactory attention of mice could be modified through their learning experiences. [ABSTRACT FROM AUTHOR]

The article presents a study on biotransformation using Escherichia coli (E. coli) expressing a cytochrome P450 (CYP) which produced a great amount of 1-octanol from n-octane after 24 hours of incubation. The initial production rate of 1-octanol was maintained throughout the entire incubation period. The results show that the functional and stable expression of a CYP in E. coli was achieved for the first time.

Pseudomonas putida T-57 was isolated from an activated sludge sample after enrichment on mineral salts basal medium with toluene as a sole source of carbon. P. putida T-57 utilizes n-butanol, toluene, styrene, m-xylene, ethylbenzene, n-hexane, and propylbenzene as growth substrates. The strain was able to grow on toluene when liquid toluene was added to mineral salts basal medium at 10–90% (v/v), and was tolerant to organic solvents whose log P ow (1-octanol/water partition coefficient) was higher than 2.5. Enzymatic and genetic analysis revealed that P. putida T-57 used the toluene dioxygenase pathway to catabolize toluene. A cis-toluene dihydrodiol dehydrogenase gene ( todD) mutant of T-57 was constructed using a gene replacement technique. The todD mutant accumulated o-cresol (maximum 1.7 g/L in the aqueous phase) when cultivated in minimal salts basal medium supplemented with 3% (v/v) toluene and 7% (v/v) 1-octanol. Thus, T-57 is thought to be a good candidate host strain for bioconversion of hydrophobic substrates in two-phase (organic-aqueous) systems. [ABSTRACT FROM AUTHOR]

Examines the biotransformation using alkane-oxidizing bacteria or their alkane hydroxylase at the molecular level. Construction of biotransformation systems for various alkanes; Details on the bacterial metabolism of normal alkanes; Importance of regio-selective oxidation of chemicals for the production of medicinal drugs.

Reports on the development of a novel bioluminescence assay of ATP by researchers from Japan. Use of ATP amplification technique on the development of the novel medical screening; Sensitivity of the bioluminescence assay on the detection of a single cell bacteria; Advantages of using the novel method over conventional bioluminescence assay in detecting bacterial contamination.

It was previously shown that the chemotaxis gene cluster 1 (cheYZABW) was required for chemotaxis. In this study, the involvement of the same cluster in aerotaxis is described and two transducer genes for aerotaxis are identified. Aerotaxis assays of a number of deletion–insertion mutants of Pseudomonas aeruginosa PAO1 revealed that the chemotaxis gene cluster 1 and cheR are required for aerotaxis. Mutant strains which contained deletions in the methyl-accepting chemotaxis protein-like genes tlpC and tlpG showed decreased aerotaxis. A double mutant deficient in tlpC and tlpG was negative for aerotaxis. TlpC has 45% amino acid identity with the Escherichia coli aerotactic transducer Aer. The TlpG protein has a predicted C-terminal segment with 89% identity to the highly conserved domain of the E. coli serine chemoreceptor Tsr. A hydropathy plot of TlpG indicated that hydrophobic membrane-spanning regions are missing in TlpG. A PAS motif was found in the N-terminal domains of TlpC and TlpG. On this basis, the tlpC and tlpG genes were renamed aer and aer-2, respectively. No significant homology other than the PAS motif was detected in the N-terminal domains between Aer and Aer-2. [Copyright &y& Elsevier]