• Title/Summary/Keyword: Bacterial metabolism

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Microorganisms Involved in Natural Fermentation of Asparagus cochinchinensis Roots and Changes in Efficacies after Fermentation (천문동 뿌리의 자연발효에 관여하는 미생물 및 발효 후 효능 변화)

  • Kim, Min-Jee;Shin, Na Rae;Lee, Myeong-Jong;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
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    • v.18 no.2
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    • pp.96-105
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    • 2018
  • Objectives: The aim of this study was to examine the effect of Asparagus cochinchinensis (AC) and fermented AC (fAC) on microorganisms and efficacies. Methods: AC was fermented for four weeks without using any bacterial strains. Then we investigated fermentation characteristics including potential of hydrogen (pH), total sugar, microbial profiling and antioxidant compound contents such as total polyphenol and total flavonoid. The anti-obesity effects of AC and fAC were evaluated by using Oil Red O staining in 3T3-L1 adipocyte. Also anti-diabetic effects of them were evaluated by using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) uptake in C2C12 skeletal muscle cell. Results: Both pH and total sugar of fAC were decreased significantly compared to unfermented AC. And the abundance of total bacteria and lactic acid bacteria increased during fermentation, especially Lactobacillus plantarum. Also fermentation of AC increased the content of total polyphenol. On the metabolic aspects, we found that AC and fAC suppressed fat accumulation. Conclusions: After four weeks of fermentation, AC increased concentrations of active compounds, altered microbial composition, and inhibited fat accumulation such as triglyceride. These results indicate that fermentation of AC might be a beneficial therapeutic approach for obesity.

Complete genome sequencing and comparative genomic analysis of Lactobacillus acidophilus C5 as a potential canine probiotics

  • Son, Seungwoo;Lee, Raham;Park, Seung-Moon;Lee, Sung Ho;Lee, Hak-Kyo;Kim, Yangseon;Shin, Donghyun
    • Journal of Animal Science and Technology
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    • v.63 no.6
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    • pp.1411-1422
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    • 2021
  • Lactobacillus acidophilus is a gram-positive, microaerophilic, and acidophilic bacterial species. L. acidophilus strains in the gastrointestinal tracts of humans and other animals have been profiled, but strains found in the canine gut have not been studied yet. Our study helps in understanding the genetic features of the L. acidophilus C5 strain found in the canine gut, determining its adaptive features evolved to survive in the canine gut environment, and in elucidating its probiotic functions. To examine the canine L. acidophilus C5 genome, we isolated the C5 strain from a Korean dog and sequenced it using PacBio SMRT sequencing technology. A comparative genomic approach was used to assess genetic relationships between C5 and six other strains and study the distinguishing features related to different hosts. We found that most genes in the C5 strain were related to carbohydrate transport and metabolism. The pan-genome of seven L. acidophilus strains contained 2,254 gene families, and the core genome contained 1,726 gene families. The phylogenetic tree of the core genes in the canine L. acidophilus C5 strain was very close to that of two strains (DSM20079 and NCFM) from humans. We identified 30 evolutionarily accelerated genes in the L. acidophilus C5 strain in the ratio of non-synonymous to synonymous substitutions (dN/dS) analysis. Five of these thirty genes were associated with carbohydrate transport and metabolism. This study provides insights into genetic features and adaptations of the L. acidophilus C5 strain to survive the canine intestinal environment. It also suggests that the evolution of the L. acidophilus genome is closely related to the host's evolutionary adaptation process.

The Regulation of LexA on UV-Induced SOS Response in Myxococcus xanthus Based on Transcriptome Analysis

  • Sheng, Duo-hong;Wang, Ye;Wu, Shu-ge;Duan, Rui-qin;Li, Yue-zhong
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.912-920
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    • 2021
  • SOS response is a conserved response to DNA damage in prokaryotes and is negatively regulated by LexA protein, which recognizes specifically an "SOS-box" motif present in the promoter region of SOS genes. Myxococcus xanthus DK1622 possesses a lexA gene, and while the deletion of lexA had no significant effect on either bacterial morphology, UV-C resistance, or sporulation, it did delay growth. UV-C radiation resulted in 651 upregulated genes in M. xanthus, including the typical SOS genes lexA, recA, uvrA, recN and so on, mostly enriched in the pathways of DNA replication and repair, secondary metabolism, and signal transduction. The UV-irradiated lexA mutant also showed the induced expression of SOS genes and these SOS genes enriched into a similar pathway profile to that of wild-type strain. Without irradiation treatment, the absence of LexA enhanced the expression of 122 genes that were not enriched in any pathway. Further analysis of the promoter sequence revealed that in the 122 genes, only the promoters of recA2, lexA and an operon composed of three genes (pafB, pafC and cyaA) had SOS box sequence to which the LexA protein is bound directly. These results update our current understanding of SOS response in M. xanthus and show that UV induces more genes involved in secondary metabolism and signal transduction in addition to DNA replication and repair; and while the canonical LexA-dependent regulation on SOS response has shrunk, only 5 SOS genes are directly repressed by LexA.

Proteome Analysis of Paenibacillus polymyxa E681 Affected by Barley

  • Seul, Keyung-Jo;Park, Seung-Hwan;Ryu, Choong-Min;Lee, Yong-Hyun;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.934-944
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    • 2007
  • Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

Differential Gene Expression of Soybean[Glycine max(L.) Merr.] in Response to Xanthomonas axonopodis pv. glycines by Using Oligonulceotide Macroarray

  • Van, Kyujung;Lestari, Puji;Park, Yong-Jin;Gwag, Jae-Gyun;Kim, Moon-Young;Kim, Dong-Hyun;Heu, Sung-Gi;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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    • pp.147-158
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    • 2007
  • Xanthomonas axonopodis pv. glycines(Xag) is a pathogen that causes bacterial leaf pustule(BLP) disease in soybeans grown in Korea and the southern United States. Typical and early symptoms of the disease are small, yellow to brown lesions with raised pustules that develop into large necrotic lesions leading to a substantial loss in yield due to premature defoliation. After Xag infects PI 96188, only pustules without chlorotic haloes were observed, indicating the different response to Xag. To identify differentially expressed genes prior to and 24 hr after Xag inoculation to PI 96188 and BLP-resistant SS2-2, an oligonucleotide macroarray was constructed with 100 genes related to disease resistance and metabolism from soybean and Arabidopsis. After cDNAs from each genotype were applied on the oligonucleotide macroarrays with three replicates and dye swapping, 36 and 81 genes were expressed as significantly different between 0 hr and 24 hr in PI 96188 and SS2-2, respectively. Six UniGenes, such as the leucine-rich repeat protein precursor or 14-3-3-like protein, were selected because they down-regulated in PI 96188 and up-regulated in SS2-2 after Xag infection, simultaneously. Using tubulin and cDNA of Jangyeobkong(BLP-susceptible) as controls, the oligonucleotide macroarray data concurred with quantitative real-time RT-PCR(QRT RT-PCR) results in most cases, supporting the accuracy of the oligonucleotide macroarray experiments. Also, QRT RT-PCR data suggested six candidate genes that might be involved in a necrotic response to Xag in PI 96188.

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Identification of Protein Kinases by Anti-phosphoserine/Phosphothreonine/Phosphotyrosine Antibody Immunoaffinity Column Chromatographies in Streptomyces griseus. (Anti-Phosphoserine/Phosphothreonine/Phesphotyrosine Antibody Immunoaffinity Column Chromatography를 이용한 Streptomyces griseus의 인산화 단백질 동정)

  • Cheong, Yong-Hoon;Kim, Jong-Hee
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.112-117
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    • 2007
  • Protein kinases play very important role for maintaining viability in prokaryote and eukaryote. The metabolism of prokaryotic cell is generally regulated by bacterial two-component regulatory systems that are composed of histidine and asparitic acid kinases, however, some eukaryotic signal transduction system such as, serine and threonine kinases, have been also found to be involved in the regulation of morphogenesis and physiological differentiation in Streptomyces. Streptomyces griseus, a streptomycin producer, was expected to have varlous types of eukaryotic-type serine/threonine protein kinases, controlling morphogenesis. Thus, many steps of chromatographies were applied to isolate serine and threonine kinases from S. griseus IFO13350. The immunoaffinity steps using anti-phosphoserine, anti-phosphothreonine, and anti-phosphotyrosine agarose column chramatographies were successfully introduced to identify eukaryotic protein kinases from S. griseus IFO13350. Eight proteins with the expected molecular weight of 14, 29, 31, 35, 40, 52, 56, and 60 kDa, were identified on SDS-PAGE, and the their kination activity was confirmed by nonradioactive protein kination assay using FITC-labeled peptide as the substrate.

Gut Microbiome Alterations and Functional Prediction in Chronic Spontaneous Urticaria Patients

  • Zhang, Xinyue;Zhang, Jun;Chu, Zhaowei;Shi, Linjing;Geng, Songmei;Guo, Kun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.747-755
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    • 2021
  • The effects of the gut microbiome on both allergy and autoimmunity in dermatological diseases have been indicated in several recent studies. Chronic spontaneous urticaria (CSU) is a disease involving allergy and autoimmunity, and there is no report detailing the role of microbiota alterations in its development. This study was performed to identify the fecal microbial composition of CSU patients and investigate the different compositions and potential genetic functions on the fecal microbiota between CSU patients and normal controls. The gut microbiota of CSU patients and healthy individuals were obtained by 16s rRNA massive sequencing. Gut microbiota diversity and composition were compared, and bioinformatics analysis of the differences was performed. The gut microbiota composition results showed that Firmicutes, Bacteroidetes, Proteobacteria, and Verrucomicrobia were dominant microbiota in CSU patients. The differential analysis showed that relative abundance of the Proteobacteria (p = 0.03), Bacilli (p = 0.04), Enterobacterales (p = 0.03), Enterobacteriaceae (p = 0.03) was significantly increased in CSU patients. In contrast, the relative abundance of Megamonas, Megasphaera, and Dialister (all p < 0.05) in these patients significantly decreased compared with healthy controls. The different microbiological compositions impacted normal gastrointestinal functions based on function prediction, resulting in abnormal pathways, including transport and metabolism. We found CSU patients exhibited gut microbiota dysbiosis compared with healthy controls. Our results indicated CSU is associated with gut microbiota dysbiosis and pointed out that the bacterial taxa increased in CSU patients, which might be involved in the pathogenesis of CSU. These results provided clues for future microbial-based therapies on CSU.

Effects of High Pressure Treatment on the Microbiological and Chemical Properties of Milk (초고압 처리가 우유의 미생물학적 및 이화학적 특성에 미치는 영향)

  • Lee, Jieun;Choi, Eun-Ji;Park, Sun Young;Jeon, Ga Young;Jang, Ja-Young;Oh, Young Jun;Lim, Seul Ki;Kim, Tae-Woon;Lee, Jong-Hee;Park, Hae Woong;Kim, Hyun Ju;Jeon, Jung Tae;Choi, Hak-Jong
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.267-274
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    • 2014
  • High pressure processing (HPP) is a non-thermal method used to prevent bacterial growth in the food industry. Currently, pasteurization is the most common method in use for most milk processing, but this has the disadvantage that it leads to changes in the milk's nutritional and chemical properties. Therefore, the effects of HPP treatment on the microbiological and chemical properties of milk were investigated in this study. With the treatment of HPP at 600 MPa and $15^{\circ}C$ for 3 min, the quantity of microorganisms and lactic acid bacteria were reduced to the level of 2-3 log CFU/ml, and coliforms were not detected during a storage period of 15 d at $4^{\circ}C$. An analysis of milk proteins, such as ${\alpha}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin by on-chip electorophoresis revealed that the electrophoretic pattern of the proteins from HPP-treated milk was different from that of conventionally treated commercial milk. While the quantities of vitamins and minerals in HPP-treated milk were seen to be comparable to amounts found in raw milk, the enzyme activity of lipase, protease and alkaline phosphatase after HPP treatment was reduced. These results suggest that HPP treatment is a viable method for the control of undesirable microorganisms in milk, allowing for minimal nutritional and chemical changes in the milk during the process.

Pink Pigmented Facultative Methylotrophic Bacteria(PPFMs): Introduction to Current Concepts (분홍색 색소를 형성하는 methylotrophic acteria(PPFMs): 최근 경향소개)

  • Munusamy, Madhaiyan;Sa, Tongmin;Kim, Jai-Joung
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.4
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    • pp.266-287
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    • 2004
  • The non infecting, plant associated bacteria have attracted increased attention for stimulating plant growth and as environmental friendly plant protecting agents. Pink-pigmented facultatively methylotrophic bacteria (PPFMs), classified as Methylobacterium spp., are persistent colonizers of plant leaf surfaces. As the leaves of most or all plants harbor PPFMs that utilize leaf methanol as their sole source of carbon and energy, which is a specific attribute of the genus Methylobacterium. Although they are not well known, these bacteria are co-evolved, interacting partners in plant metabolism. This claim is supported, for example, by the following observations: (1) PPFMs are seed-transmitted, (2) PPFMs are frequently found in putatively axenic cell cultures, (3) Low numbers of seed-borne PPFMs correlate with low germinability, (4) Plants with reduced numbers of PPFM show elevated shoot/root ratios, (5) Foliar application of PPFMs to soybean during pod fill enhances seed set and yield, (6) Liverwort tissue in culture requires PPFM-produced vitamin B12 for growth, (7) treated plants to suppress or decrease disease incidence of sheath blight caused by Rhizoctonia solani in rice, and (8) the PPFM inoculation induced number of stomata, chlorophyll concentration and malic acid content, they led to increased photosynthetic activity. Methylobacterium spp. are bacterial symbionts of plants, shown previously to participate in plant metabolism by consuming plant waste products and producing metabolites useful to the plant. There are reports that inform about the beneficial interactions between this group of bacteria and plants. Screening of such kind of bacteria having immense plant growth promoting activities like nitrogen fixation, phytohormone production, alleviating water stress to the plants can be successfully isolated and characterized and integration of such kind of organism in crop production will lead to increased productivity.

Influence of Temperature and pH on Fermentation Pattern and Methane Production in the Rumen Simulating Fermenter (RUSITEC)

  • Bhatta, R.;Tajima, K.;Kurihara, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.376-380
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    • 2006
  • An experiment was conducted to study the effect of temperature and pH on in vitro nutrient degradability, volatile fatty acid profile and methane production. The fermenter used was the semi-continuous system, known as the rumen simulation technique (RUSITEC). Sixteen cylinders were used at one time with a volume of 800 ml, the dilution rate was set at 3.5%/hour, the infused buffer being McDougall's artificial saliva. Basal diet (9.6 g DM) used in RUSITEC consisted of (DM) 6.40 g Timothy hay, 1.86 g crushed corn and 1.34 g soybean meal. The food for the fermentation vessel was provided in nylon bags, which were gently agitated in the liquid phase. The experiment lasted for 17 d with all the samples taken during the last 5 d. Treatments were allocated at random to four vessels each and were (1) two temperature levels of $39^{\circ}C$ and $41^{\circ}C$ (2) two pH levels of 6.0 and 7.0. The total diet contained ($g\;kg^{-1}$ DM) 957 OM, 115 CP and $167MJ\;kg^{-1}$ (DM) GE. Although increase in temperature from $39^{\circ}C$ to $41^{\circ}C$ reduced degradation of major nutrients in vitro, it was non-significant. Interaction effect of temperature with pH also reflected a similar trend. However, pH showed a significant (p<0.05) negative effect on the degradability of all the nutrients in vitro. Altering the in vitro pH from 7 to 6 caused marked reduction in DMD from 60.2 to 41.8, CPD from 76.3 to 55.3 and GED from 55.3 to 35.1, respectively. Low pH (6) depressed total VFA production (61.9 vs. 34.9 mM) as well as acetate to propionate ratio in vitro (from 2.0 to 1.5) when compared to pH 7. Compared to pH 7, total gas production decreased from 1,841 ml to 1,148 ml at pH 6, $CO_2$ and $CH_4$ production also reduced from 639 to 260 ml and 138 to 45 ml, respectively. This study supported the premise that pH is one of the principal factors affecting the microbial production of volatile fatty acids and gas. Regulating the ruminal pH to increase bacterial activity may be one of the methods to optimize VFA production, reduce methane and, possibly, improve animal performance.