Uncovering potential interspecies signaling factors in plant-derived mixed microbial culture.

Microbes use signaling factors for intraspecies and interspecies communications. While many intraspecies signaling factors have been found and characterized, discovery of factors for interspecies communication is lagging behind. To facilitate the discovery of such factors, we explored the potential of a mixed microbial culture (MMC) derived from wheatgrass, in which heterogeneity of this microbial community might elicit signaling factors for interspecies communication. The stability of Wheatgrass MMC in terms of community structure and metabolic output was first characterized by 16S ribosomal RNA amplicon sequencing and liquid chromatography/mass spectrometry (LC/MS), respectively. In addition, detailed MS analyses led to the identification of 12-hydroxystearic acid (12-HSA) as one of the major metabolites produced by Wheatgrass MMC. Stereochemical analysis revealed that Wheatgrass MMC produces mostly the (R)-isomer, although a small amount of the (S)-isomer was also observed. Furthermore, 12-HSA was found to modulate planktonic growth and biofilm formation of various marine bacterial strains. The current study suggests that naturally derived MMCs could serve as a simple and reproducible platform to discover potential signaling factors for interspecies communication. In addition, the study indicates that hydroxylated long-chain fatty acids, such as 12-HSA, may constitute a new class of interspecies signaling factors.

Sequential evolution of bacterial morphology by co-option of a developmental regulator.

What mechanisms underlie the transitions responsible for the diverse shapes observed in the living world? Although bacteria exhibit a myriad of morphologies, the mechanisms responsible for the evolution of bacterial cell shape are not understood. We investigated morphological diversity in a group of bacteria that synthesize an appendage-like extension of the cell envelope called the stalk. The location and number of stalks varies among species, as exemplified by three distinct subcellular positions of stalks within a rod-shaped cell body: polar in the genus Caulobacter and subpolar or bilateral in the genus Asticcacaulis. Here we show that a developmental regulator of Caulobacter crescentus, SpmX, is co-opted in the genus Asticcacaulis to specify stalk synthesis either at the subpolar or bilateral positions. We also show that stepwise evolution of a specific region of SpmX led to the gain of a new function and localization of this protein, which drove the sequential transition in stalk positioning. Our results indicate that changes in protein function, co-option and modularity are key elements in the evolution of bacterial morphology. Therefore, similar evolutionary principles of morphological transitions apply to both single-celled prokaryotes and multicellular eukaryotes.

Efficient microbial conversion of L-tyrosine to L-DOPA by Brevundimonas sp. SGJ.

L-DOPA (3,4-dihydroxyphenyl-L-alanine), the most widely used drug for the treatment of Parkinson's disease, was produced in buffer using biomass of Brevundimonas sp. SGJ. The effects of enhancers, such as carrageenan, diatomaceous earth, and activated charcoal, on the L-DOPA production were evaluated to obtain the maximum yield. The optimal process conditions found were pH 8, 2 g l⁻¹ cell mass, 2 g l⁻¹ L-tyrosine, 0.04 g l⁻¹ CuSO4, 0.02 g l⁻¹ L-ascorbic acid, 0.5 g l⁻¹ carrageenan, and 40 °C temperature. In addition, repeated use of cells resulted in the highest yield of 3.81 g l⁻¹ (95.2%) of L-DOPA with utilization of 4 g l⁻¹ L-tyrosine, and the highest tyrosinase activity (9,201 U mg⁻¹) was observed at 18 h of incubation. Furthermore, the produced L-DOPA was confirmed by high-performance thin-layer chromatography, high-performance liquid chromatography, and gas chromatography-mass spectroscopy. Kinetic studies showed significant values of Y (p/s), Q (s), and q (s) after optimization of the process. Thus, Brevundimonas sp. SGJ could be an eventual new source for large-scale production of L-DOPA.

Phenylobacterium falsum sp. nov., an Alphaproteobacterium isolated from a nonsaline alkaline groundwater, and emended description of the genus Phenylobacterium.

A Gram-negative bacterium designated AC-49T was isolated from an alkaline groundwater with a pH 11.4. This organism formed rod-shaped cells, was strictly aerobic, catalase and oxidase positive, with an optimum growth temperature of 35 degrees C and an optimum pH value of 8.0. Strain AC-49T assimilated primarily amino acids and some Krebs cycle metabolites, did not use sugars for growth. The organism did not grow on L-phenylalanine or antipyrin. The G+ C content of DNA was 66.9 mol%. The phylogenetic analyses based on the 16S rRNA sequencing showed that the closest relatives of strain AC-49T were Phenylobacterium lituiforme and Phenylobacterium immobile, indicating that the organism is a member of the order Caulobacterales of the Alphaproteobacteria. Based on the phylogenetic analyses and distinct phenotypic characteristics, we are of the opinion that strain AC-49T, represents a novel species of the genus Phenylobacterium for which we propose the name Phenylobacterium falsum sp. nov.

Asticcacaulis taihuensis sp. nov., a novel stalked bacterium isolated from Taihu Lake, China.

A novel stalked bacterium, designated strain T3-B7(T), was isolated from sediment of Taihu Lake, Jiangsu Province, China, and its taxonomy was studied by using a polyphasic approach. Cell morphology, physiological and biochemical properties, and polar lipids indicated that strain T3-B7(T) represented a member of the genus Asticcacaulis. Based on 16S rRNA gene sequence similarity analysis, strain T3-B7(T) was found to be phylogenetically related to Asticcacaulis biprosthecium DSM 4723(T) (98.5 %) and Asticcacaulis excentricus DSM 4724(T) (95.0 %), but could be differentiated from these two species on the basis of the number and position of prosthecae, assimilation of sugars, nitrate reduction and tolerance to NaCl. Levels of DNA-DNA relatedness of strain T3-B7(T) to A. biprosthecium DSM 4723(T) and A. excentricus DSM 4724(T) were 37.1 and 18.0 %, respectively. The G + C content of strain T3-B7(T) was 59 mol% (T(m)). It is concluded that strain T3-B7(T) represents a novel species of the genus Asticcacaulis, for which the name of Asticcacaulis taihuensis sp. nov. is proposed. The type strain is T3-B7(T) (=AS 1.3431(T) = JCM 12463(T)).

Phenylobacterium lituiforme sp. nov., a moderately thermophilic bacterium from a subsurface aquifer, and emended description of the genus Phenylobacterium.

A facultative anaerobic bacterium, strain FaiI3(T), was isolated from samples collected from the free-flowing waters of a bore well (Fairlea Bore, registration number 3768) which taps into the Australian Great Artesian Basin subsurface thermal aquifer. Strain FaiI3(T) developed yellow to pale-yellow colonies (0.5-1.5 mm) after 48 h. The non-spore forming rods (0.5x1-3 microm) were slightly curved, occurred singly and as pairs and were motile with a single polar flagellum. Cells tended to form clumps in liquid medium and rosettes were commonly observed. The cells stained Gram-negative and electron micrographs of thin sections revealed a multi-layered complex Gram-negative cell wall structure. Strain FaiI3(T) grew optimally at 40-41 degrees C, with growth observed at 45 degrees C but not at 50 degrees C. The pH growth range was between pH 6 and 9 and optimal growth occurred between pH 6 and 6.5. Strain FaiI3(T) grew best with yeast extract as the sole carbon and energy source. Peptone, yeast extract, acetate, xylose, sucrose, glucose, glycerol, succinate, butyrate, lactate, fumarate, citrate, L-phenylalanine, cellobiose and gelatin supported growth but maltose, fructose, glycine, ethanol, benzoate and oxalate did not. Tyrosine was produced from L-phenylalanine. Strain FaiI3(T) was catalase-positive and oxidase-negative and did not hydrolyse starch. Growth was inhibited by neomycin, tetracycline, streptomycin, chloramphenicol, ampicillin, vancomycin and spectinomycin. The G+C content was determined to be 66.5+/-0.5 mol%. On the basis of the 16S rRNA gene sequence analysis, strain FaiI3(T) was assigned as a novel species of the genus Phenylobacterium, Phenylobacterium lituiforme sp. nov. in the order Caulobacterales, subclass alpha-Proteobacteria, class Proteobacteria. The type strain is FaiI3(T) (=ATCC BAA-294(T)=DSM 14363(T)).