Journal of General and Applied Microbiology最新文献

Mycoplasma pneumoniae is one of the most important pathogens causing community acquired pneumonia in children, and the pathogenic mechanism of M. pneumoniae infection is complex. Azithromycin is an effective agent for treating the acquired lower respiratory tract infection and urogenital tract infection with slight adverse reactions. This study aimed to compare the intestinal microflora before (PP1) and after azithromycin intervention (PP2) in children with pneumonia caused by M. pneumoniae, combined with body fluid biochemical analysis to determine the intestinal flora affecting the progress of the disease. Fifteen children diagnosed with M. pneumoniae pneumonia were recruited. The fecal samples and clinical biochemical data were collected. 16S rRNA gene amplicon sequencing and bioinformatics analysis were conducted by the Beijing Genomics Institute. The operational taxonomic unit abundance analysis showed significant differences between the two groups. The species richness analysis showed differences in class, family, genus, order, species, and phylum. The abundance of Haemophilus, Pasteurellales, and Pasteurellaceae was found to be significantly higher in the PP1 group. The Pearson correlation analysis showed that the microbes strongly correlated with the clinical features. 16S rRNA gene sequencing data revealed altered composition of gut microbiota in children with M. pneumoniae pneumonia treated with azithromycin. The altered expression of microbes correlated with clinical features, which might help diagnose and treat the disease.

From the biotechnological point of view, enzymes are powerful tools that help sustain a clean environment in several ways. The enzymatic biodegradation of synthetic dyes is a promising goal since it reduces pollution caused by textile dyeing factory wastewater. Lignin peroxidase (EC 1.11.1.14, LiP) has high redox potential; thus, it is great for application in various industrial fields (e.g., paper- waste treatment and textile dyeing wastewater treatment). In the present study, a LiP from an isolated strain Pleurotus pulmonarius CPG6 (PpuLiP) was successfully purified with a specific activity of 6.59 U mg ^-1. The enzyme was purified by using three-step column chromatography procedures including DEAE, Sephadex G-75, and HiTrap^TM Q FF columns with 17.8-fold purity. The enzyme with a molecular weight of 40 kDa exhibited enhanced pH stability in the acidic range. The activity retention was over 75% at a pH of 3.0 for more than 6 hours. Purified PpuLiP was able to oxidize a variety of substrates including veratryl alcohol, 2,4-DCP, n propanol, and guaiacol. The effect of metal ions on PpuLiP activity was analyzed. The study will provide a ground to decolorize dyes from various groups of PpuLiP. Purified PpuLiP could decolorize 35% Acid blue 25 (AB25), 50% Acid red 129 (AB129), 72% Acid blue 62 (NY3), 85% Acid blue 113 (AB113), 55% Remazol Brilliant blue R (RBBR), and 100% Reactive red 120 (RR120) for 12 hours. Most of the dyes were decolorized, but the heat-denatured enzyme used as negative control obviously did not decolorize the tested dyes. These results indicate that the PpuLiP has potential application in enzyme-based decolorization of synthetic dyes. Keywords: Decolorization; lignin peroxidase; Pleurotus pulmonarius; textile dyes.

Fission yeast, Schizosaccharomyces pombe, possesses eight hexose transporters, Ght1~8. In order to clarify the role of each hexose transporter on glucose uptake, a glucose uptake assay system was established and the actual glucose uptake activity of each hexose transporter-deletion mutant was measured. Under normal growth condition containing 2% glucose, ∆ght5 and ∆ght2 mutants showed large and small decrease in glucose uptake activity, respectively. On the other hand, the other deletion mutants did not show any decrease in glucose uptake activity indicating that, in the presence of Ght5 and Ght2, the other hexose transporters do not play a significant role in glucose uptake. To understand the relevance between glucose uptake and lifespan regulation, we measured the chronological lifespan of each hexose transporter deletion mutant, and found that only ∆ght5 mutant showed a significant lifespan extension. Based on these results we showed that Ght5 is mainly involved in the glucose uptake in Schizosaccharomyces pombe, and suggested that the ∆ght5 mutant has prolonged lifespan due to physiological changes similar to calorie restriction.

Glucuronoyl esterase (GE) is a promising agent for the delignification of plant biomass since it has been shown to cleave the linkage between xylan and lignin in vitro. In this study, we demonstrate that NcGE, a GE from Neurospora crassa, stimulates plant biomass degradation. In vitro, NcGE synergistically increased the release of reducing sugars from plant biomass when added together with cellulase or xylanase. In vivo, overexpression of NcGE in N. crassa resulted in an increase in xylanolytic activity. Consistently, elevated transcription of genes encoding the major plant biomass degrading-enzymes (PBDEs) was observed in the NcGE overexpression strain. Increased xylanolytic activity and transcription of PDBE genes were largely abolished when the transcription factors clr-1, clr-2, or xlr-1 were deleted. Interestingly, the expression of some PBDE genes was increased when the hydrolysate of plant biomass by NcGE was added to the culture medium. We propose that NcGE boosts the production of PBDEs through the activation of key transcription factors, which is presumably caused by NcGE-mediated generation of hypothetical inducer(s) from plant biomass.

In Saccharomyces cerevisiae, ethyl caprylate is produced by the esterification of caprylic acid, which is synthesized through the action of fatty acid synthase. A recent study reported a yeast mutant with a single nucleotide substitution in the alpha subunit of fatty acid synthase (FAS2) gene (F1279Y; 3836T>A) that produced large amounts of ethyl caprylate. Here, we designed two primer sets (P1/P2 and P3/P4) with mismatches that incorporate restriction sites for the enzymes NdeI and SspI, respectively and developed an easy and rapid polymerase chain reaction-restriction fragment length polymorphism assay to identify yeasts harboring the FAS2-F1279Y mutation associated with high ethyl caprylate productivity.

As a central signaling molecule, c-di-GMP (bis-(3,5)-cyclic diguanosine monophosphate) is becoming the focus for research in bacteria physiology. Pseudomonas aeruginosa PAO1 genome contains highly complicated c-di-GMP metabolizing genes and a number of these proteins have been identified and investigated. Especially, a sophisticated network of these proteins is emerging. In current study, mainly through Bacteria-2-Hybrid assay, we found PA0575 (RmcA), a GGDEF-EAL dual protein, to interact with two other dual proteins of PA4601 (MorA) and PA4959 (FimX). These observations imply the intricacy of c-di-GMP metabolizing protein interactions. Our work thus provides one piece of data to increase the understandings to c-di-GMP signaling.

The decolorization of 11 dyes by granular sludge from an anaerobic expanded granular sludge bed (EGSB) reactor was evaluated. Biological decolorization of Reactive Red 21, 23, and 180, and Reactive Yellow 15, 17, and 23 in model textile wastewater was observed for the first time after a 7-day incubation (over 94% decolorization). According to the sequencing analysis of 16S rRNA gene amplicons from EGSB granular sludge, the operational taxonomic unit related to Paludibacter propionicigenes showed the highest increase in relative abundance ratios in the presence of dyes (7.12 times on average over 11 dyes) compared to those without dyes.

Consumption of temperature-abused marine fish containing elevated levels of histamine results in histamine poisoning. Histamine is a biogenic amine produced in fish by the action of certain groups of bacteria which are capable of producing an exogenous enzyme called histidine decarboxylase (HDC). Morganella morganii is one of the major causative organisms of histamine poisoning. In this study, the histamine forming potential of M. morganii (BSS142) was evaluated when it was co-incubated with proteolytic as well as polyamine forming bacteria. This experiment was designed to examine whether biotic factors such as proteolysis and the presence of other amines influenced histamine forming ability of BSS142. The study showed that the proteolytic activity of Aeromonas hydrophila as well as Pseudomonas aeruginosa greatly enhanced the histamine forming ability of M. morganii. Psychrobacter sangunis, a non proteolytic polyamine producer, negatively influenced histamine production by M. morganii.

Godo is a traditional fermented soy food made in Aomori prefecture, Japan. It is mainly made of soybeans, rice koji, and salt. Since godo ripens during the long and severe winter in northeast Japan, it is assumed that lactic acid bacteria inhabiting godo have cold tolerance. We aimed to investigate the presence or absence of psychrotrophic lactic acid bacteria in godo. The viable counts of estimated lactic acid bacteria ranged from 10⁶ to 10⁸ cfu/g. In addition, aerobic and anaerobic microorganisms were detected in four godo products though the microbial population differed from sample to sample. Twenty-two bacterial strains were able to be isolated from godo, and all of the isolated strains were Gram-positive and catalase-negative. Some of the isolates grew well at 10°C. The carbohydrate fermentation profile of the selected three strains was determined by API50 CHL analysis. These strains were identified as Leuconostoc mesenteroides, and Latilactobacillus sakei by 16S rRNA gene sequence analysis. Leuconostoc mesenteroides strains HIT231 and HIT252, and Latilactobacillus sakei strain HIT273 could grow at 5°C in MRS broth, but their optimum growth temperature was 20°C-30°C. These results suggest that psychrotrophic lactic acid bacteria presumed to be derived from rice koji are present in godo, which is one of the factors in the low temperature ripening of godo in winter.

The present study investigated the efficacy of bacterial cellulose production by K. xylinus TISTR 1011 and K. nataicola TISTR 975 using yam bean juice as a nutrient source, and the physicochemical and sensory characteristics of bacterial cellulose were examined. Bacterial cellulose content, production yield, and production rate were significantly higher when K. xylinus TISTR 1011 rather than K. nataicola TISTR 975 was used as the bacterial strain. The analysis of physicochemical characteristics revealed that bacterial cellulose produced by K. xylinus TISTR 1011 using yam bean juice medium had higher scores for CIE L*, a*, and b* values, wet weight, moisture content, firmness, and gel strength than bacterial cellulose produced by K. nataicola TISTR 975. In contrast, sensory evaluation showed that the acceptability scores and preference of all attributes of bacterial cellulose produced by K. nataicola TISTR 975 using yam bean juice medium were higher than those of bacterial cellulose produced by K. xylinus TISTR 1011. The results of this study indicate that yam bean juice from yam bean tubers, an alternative raw material agricultural product, can be used as a nutrient source for producing bacterial cellulose or nata by Komagataeibacter strains.