• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.470-478
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• 2016

A bacterium producing a large amount of chitinolytic enzyme was isolated from the intestinal tract of earthworm. The isolate was identified as Bacillus licheniformis by 16S ribosomal RNA analysis and designated as B. licheniformis GA9. The enzyme was purified by 40-60% ammonium sulfate precipitation, diethyl-aminoethyl groups exchange chromatography, and gel filtration chromatography. The molecular weight was estimated to be 52.1 kDa and the N-terminal amino acid sequence was D-S-G-K-N-G-K-I-I-R-Y-YP-I-R. The optimum activity of the purified chitinolytic enzyme was shown at pH 5.0 and $40^{\circ}C$, and the enzyme was stable in the ranges of $20-50^{\circ}C$ and pH 5.0-6.0. Enzyme activity was increased by $Co^{2+}$, while it was inhibited by $Cu^{2+}$ and $Fe^{2+}$. But it was recovered by chelating metals with ethylenediaminetetraacetic acid. The $K_m$ and $V_{max}$ values of the purified enzyme were 4.02 mg/ml and 0.52 mg/min, respectively. The chitinolytic enzyme characterized in this study has potential applications in areas such as biotechnology, biomedicine, agriculture, and nutrition.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.5
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• pp.275-282
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• 2005

By using differential display, we identified one of the genes encoding the multi-subunit complex protein V-ATPase, c subunit gene (ATP6L), and showed alterations of the gene expression by oxidative stresses. Expression of the ATP6L gene in Neuro-2A cells was increased by the treatment with $H_2O_2$ and incubation in hypoxic chamber, implying that the expression of the ATP6L gene is regulated by oxidative stresses. To examine mechanisms involved in the regulation of the gene expression by oxidative stresses, the transcriptional activity of the rat ATP6L promoter was studied. Transcription initiation site was determined by primer extension analysis and DNA sequencing, and promoter of the rat ATP6L and its deletion clones were constructed in reporter assay vector. Significant changes of the promoter activities in Neuro-2A cells were observed in two regions within the proximal 1 kbp promoter, and one containing a suppressor was in -195 to -220, which contains GC box that is activated by binding of Sp1 protein. The suppression of promoter activity was lost in mutants of the GC box. We confirmed by electrophoretic mobility shift and supershift assays that Sp1 protein specifically binds to the GC box. The promoter activity was not changed by the $H_2O_2$ treatment and incubation in hypoxic chamber, however, $H_2O_2$ increased the stability of ATP6L mRNA. These data suggest that the expression of the ATP6L gene by oxidative stresses is regulated at posttranscriptional level, whereas the GC box is important in basal activities of the promoter.

• Journal of Microbiology and Biotechnology
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• v.30 no.4
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• pp.505-514
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• 2020

The symbiotic nature of the relationship between algae and marine bacteria is well-studied among the complex microbial interactions. The mutual profit between algae and bacteria occurs via nutrient and vitamin exchange. It is necessary to analyze the genome sequence of a bacterium to predict its symbiotic relationships. In this study, the genome of a marine bacterium, Pseudoruegeria sp. M32A2M, isolated from the south-eastern isles (GeoJe-Do) of South Korea, was sequenced and analyzed. A draft genome (91 scaffolds) of 5.5 Mb with a DNA G+C content of 62.4% was obtained. In total, 5,101 features were identified from gene annotation, and 4,927 genes were assigned to functional proteins. We also identified transcription core proteins, RNA polymerase subunits, and sigma factors. In addition, full flagella-related gene clusters involving the flagellar body, motor, regulator, and other accessory compartments were detected even though the genus Pseudoruegeria is known to comprise non-motile bacteria. Examination of annotated KEGG pathways revealed that Pseudoruegeria sp. M32A2M has the metabolic pathways for all seven vitamin Bs, including thiamin (vitamin B1), biotin (vitamin B7), and cobalamin (vitamin B12), which are necessary for symbiosis with vitamin B auxotroph algae. We also identified gene clusters for seven secondary metabolites including ectoine, homoserine lactone, beta-lactone, terpene, lasso peptide, bacteriocin, and non-ribosomal proteins.

• Journal of Animal Science and Technology
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• v.44 no.1
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• pp.13-22
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• 2002

To identify the differentially expressed genes at growth stage of Hanwoo, we constructed the subtractive cDNA library from loin mRNA of 12- and 24-month old Hanwoo by PCR-based subtraction. The fourteen genes were confirmed by sequencing and reverse northern blot analysis, and they were selected as candidate of putative genes differentially expressed at the growth stage of Hanwoo. Three subtracted cDNA fragments that expressed specific signal to cDNA probe for 6-month-old loin of Hanwoo were highly homologous to those of the genes encoding EPV 20, Ca2＋ATPase, and TCTP, respectively. The nine cDNA clones showed intense signal to cDNA probe from 12-month-old loin of Hanwoo, and highly homologus to those of genes encoding VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase, and UCP 2, respectively. Two subtracted cDNA clones that expressed specific signal to cDNA probes for 12- and 24-month-old loin of Hanwoo were detected. One of them was highly homologus to the gene encoding ferrochelatase and the other was highly homologus to the gene encoding ADRP.

• Journal of Applied Biological Chemistry
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• v.54 no.1
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• pp.21-25
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• 2011

A lactic acid bacterial strain showing high acid production in saccharified-rice suspension was isolated from Kimchi. This strain was analyzed by API 50 CHL kit and 16S rRNA sequencing analysis and identified as Leuconostoc mesenteroides 310-12. Saccharified-rice suspension was fermented using L. mesenteroides 310-12 strain at $30^{\circ}C$ for 15 h. The changes of pH, titratable acidity and viable cell number during fermentation were determined. The pH and titratable acidity were reached to pH 3.57 and 0.40% after 15 h fermentation, respectively. The viable cell population of L. mesenteroides 310-12 was rapidly increased to $8.9{\times}10^8$ CFU/g during the 15 h of cultivation. The contents of lactic acid and acetic acid were determined to be 0.077% and 0.065% after 15 h fermentation, respectively. The rice-based fermented beverage was manufactured by blending L. mesenteroides 310-12 fermented broth and some food additives. When this beverage was stored at $4^{\circ}C$, the viable cells population was decreased to $1.0{\times}10^7$ CFU/g and pH was nearly maintained for 25 days.

• Journal of Microbiology and Biotechnology
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• v.28 no.11
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• pp.1883-1895
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• 2018

Alcohol dependence is a global public health problem, yet the mechanisms of alcohol dependence are incompletely understood. The traditional view has been that ethanol alters various neurotransmitters and their receptors in the brain and causes the addiction. However, an increasing amount of experimental evidence suggests that gut microbiota also influence brain functions via gut-to-brain interactions, and may therefore induce the development of alcohol use disorders. In this study, a rat model of alcohol dependence and withdrawal was employed, the gut microbiota composition was analyzed by high-throughput 16S rRNA gene sequencing, and the metagenome function was predicted by PICRUSt software. The results suggested that chronic alcohol consumption did not significantly alter the diversity and richness of gut microbiota in the jejunum and colon, but rather markedly changed the microbiota composition structure in the colon. The phyla Bacteroidetes and eight genera including Bacteroidales S24-7, Ruminococcaceae, Parabacteroides, Butyricimonas, et al were drastically increased, however the genus Lactobacillus and gauvreauii in the colon were significantly decreased in the alcohol dependence group compared with the withdrawal and control groups. The microbial functional prediction analysis revealed that the proportions of amino acid metabolism, polyketide sugar unit biosynthesis and peroxisome were significantly increased in the AD group. This study demonstrated that chronic alcohol consumption has a dramatic effect on the microbiota composition structure in the colon but few effects on the jejunum. Inducement of colonic microbiota dysbiosis due to alcohol abuse seems to be a factor of alcohol dependence, which suggests that modulating colonic microbiota composition might be a potentially new target for treating alcohol addiction.

• Journal of Life Science
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• v.30 no.6
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• pp.522-531
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• 2020

The aim of this study was to establish the optimal medium composition for enhancing L(+)-lactic acid (LLA) production using response surface methodology (RSM). Lactobacillus paracasei SRCM201474 was selected as the LLA producer by productivity analysis from nine candidates isolated from kimchi and identified by 16S rRNA gene sequencing. Plackett-Burman design was used to assess the effect of eleven media components on LLA production, including carbon (glucose, sucrose, molasses), nitrogen (yeast extract, peptone, tryptone, beef extract), and mineral (NaCl, K₂HPO₄, MgSO₄, MnSO₄) materials. Glucose, sucrose, molasses, and peptone were subsequently chosen as promising media for further optimization studies, and a hybrid design experiment was used to establish their optimal concentrations as glucose 15.48 g/l, sucrose 16.73 g/l, molasses 39.09 g/l, and peptone 34.91 g/l. The coefficient of determination of the equation derived from RSM regression for LLA production was mathematically reliable at 0.9969. At optimum parameters, 33.38 g/l of maximum LLA increased by 193% when compared with MRS broth as unoptimized medium (17.66 g/l). Our statistical model was confirmed by subsequent validation experiments. Increasing the performance of LLA-producing microorganisms and establishing an effective LLA fermentation process can be of particular benefit for bioplastic technologies and industrial applications.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.350-350
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• 2017

The study was conducted to evaluate differences of growth characteristics of rice cultivated in two different regions (Cheonan and Jeonju). It focused on nutritional composition and anti-nutritional factors of rice straw produced from 21 rice varieties including GM rice (Iksan 483). The range of general nutrition ingredient is that crude was 0.97 ~ 3.2 %, carbohydrate was 67.45 ~ 80.01 %, crude protein was 1.46 ~ 4.81 %, crude ash was 6.52 ~ 18.96 %, crude fiber was 25.77 ~ 40.02 %, NDF was 51.84 ~ 67.77 %, ADF was 27.11 ~ 40.44 %, calcium was 0.49 ~ 5.18 mg/g and phosphorous was 0.26 ~ 2.77 mg/g. The general nutritional contents of GM rice were included above range. The range of phytic acid of rice straws cultivated in Cheonan and Jeonju was 0 ~ 0.056 mg/ml and 0 ~ 0.059 mg/ml, respectively. The phytic acid content of GM was 0.033 mg/ml, which was in the range of the content of rice straw in Cheonan and Jeonju. The range of trypsin inhibitor of rice straws cultivated in Cheonan and Jeonju was 0.061 ~ 0.461 TIU/mg and 0 ~ 1.278 TIU/mg, respectively. The trypsin acid content of GM was 0.461 TIU/mg, which was in the range of the content of rice straw in Cheonan and Jeonju. In addition, we investigated microbial community from each soil sample by using metagenomics sequencing based on rRNA microbial diversity in order to inspect indirect changes of soil environment with cultivation of GM rice. Metagenomics analysis was carried out using soil samples cultivated with GM and non-GM rice for before transplanting, young panicle differentiation stage, heading stage, and ripening stage. Beta diversity of microbial community in both soil environments were calculated by using Bray-Curtis distance method and showed low value with an average of 0.24 (dissimilarity = 1). As a result, it was confirmed that the cultivation of GM does not give a significant effect on the change of microbial composition in soil. Therefore, Our study demonstrates that there is no difference in the composition of soil microorganism due to GM and non-GM rice.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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• pp.463-469
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• 2013

Cluster of differentiation 4 (CD4) is mainly expressed on $CD4^+$ T cells, which plays an important role in immune response. The aim of this study was to detect the association between polymorphisms of the CD4 gene and T lymphocyte subpopulations in pigs, and to investigate the effects of genetic variation on the CD4 gene expression level in immune tissues. Five missense mutations in the CD4 gene were identified using DNA pooling sequencing assays, and two main haplotypes (CCTCC and AGCTG) in strong linkage disequilibrium (with frequencies of 50.26% and 46.34%, respectively) were detected in the population of Large White pigs. Our results indicated that the five SNPs and the two haplotypes were significantly associated with the proportions of $CD4^-CD8^-$, $CD4^+CD8^+$, $CD4^+CD8^-$, $CD4^+$ and $CD4^+/CD8^+$ in peripheral blood (p<0.05). Gene expression analysis showed the mRNA level of the CD4 gene in thymus was significantly higher than that in lymph node and spleen (p<0.05). However, no significant difference was observed between animals with CCTCC/CCTCC genotype and animals with AGCTG/AGCTG genotype in the three immune tissues (p>0.05). These results indicate that the CD4 gene may influence T lymphocyte subpopulations and can be considered as a candidate gene affecting immunity in pigs.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.11
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• pp.1725-1731
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• 2020

Objective: An initial RNA-Sequencing study revealed that UDP-galactose-4-epimerase (GALE) was one of the most promising candidates for milk protein concentration in Chinese Holstein cattle. This enzyme catalyzes the interconversion of UDP-galactose and UDP-glucose, an important step in galactose catabolism. To further validate the genetic effect of GALE on milk protein traits, genetic variations were identified, and genotypes-phenotypes associations were performed. Methods: The entire coding region and the 5'-regulatory region (5'-UTR) of GALE were re-sequenced using pooled DNA of 17 unrelated sires. Association studies for five milk production traits were performed using a mixed linear animal model with a population encompassing 1,027 Chinese Holstein cows. Results: A total of three variants in GALE were identified, including two novel variants (g.2114 A>G and g.2037 G>A) in the 5'-UTR and one previously reported variant (g.3836 G>C) in an intron. All three single nucleotide polymorphisms (SNPs) were associated with milk yield (p<0.0001), fat yield (p = 0.0006 to <0.0001), protein yield (p = 0.0232 to <0.0001) and protein percentage (p<0.0001), while no significant associations were detected between the SNPs and fat percentage. A strong linkage disequilibrium (D' = 0.96 to 1.00) was observed among all three SNPs, and a 5 Kb haplotype block involving three main haplotypes with GAG, AGC, and AGG was formed. The results of haplotype association analyses were consistent with the results of single locus association analysis (p<0.0001). The phenotypic variance ratio above 3.00% was observed for milk protein yield that was explained by SNP-g.3836G >C. Conclusion: Overall, our findings provided new insights into the polymorphic variations in bovine GALE gene and their associations with milk protein concentration. The data indicate their potential uses for marker-assisted breeding or genetic selection schemes.