• Journal of Microbiology and Biotechnology
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• v.28 no.1
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• pp.145-156
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• 2018

Most eukaryotic peroxiredoxins (Prxs) are readily inactivated by a high concentration of hydrogen peroxide ($H_2O_2$) during catalysis owing to their "GGLG" and "YF" motifs. However, such oxidative stress sensitive motifs were not found in the previously identified filamentous fungal Prxs. Additionally, the information on filamentous fungal Prxs is limited and fragmentary. Herein, we cloned and gained insight into Aspergillus nidulans Prx (An.PrxA) in the aspects of protein properties, catalysis characteristics, and especially $H_2O_2$ tolerability. Our results indicated that An.PrxA belongs to the newly defined family of typical 2-Cys Prxs with a marked characteristic that the "resolving" cysteine ($C_R$) is invertedly located preceding the "peroxidatic" cysteine ($C_P$) in amino acid sequences. The inverted arrangement of $C_R$ and $C_P$ can only be found among some yeast, bacterial, and filamentous fungal deduced Prxs. The most surprising characteristic of An.PrxA is its extraordinary ability to resist inactivation by extremely high concentrations of $H_2O_2$, even that approaching 600 mM. By screening the $H_2O_2$-inactivation effects on the components of Prx systems, including Trx, Trx reductase (TrxR), and Prx, we ultimately determined that it is the robust filamentous fungal TrxR rather than Trx and Prx that is responsible for the extreme $H_2O_2$ tolerence of the An.PrxA system. This is the first investigation on the effect of the electron donor partner in the $H_2O_2$ tolerability of the Prx system.

• Biomedical Science Letters
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• v.12 no.2
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• pp.73-79
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• 2006

Nebulin is a giant ($600{\sim}900$ kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin filamints in vertebrate skeletal muscles. Recently, There is an evidence that the nebulin also expressed in non muscle tissue, brain and liver. We identified a new isoform of nebulin from adult brain library by PCR screening. It contains two simple-repeats exon 165, 166 and linker-repeats exon $154{\sim}161$ except exon 159. The nebulin modules M160 to M170 (exon 150 to exon 161) has been shown to bind desmin. In mature striated muscle, desmin intermediate filaments surround Z-discs and link individual myofibrils laterally at their Z-discs and to other intracellular structures, including the costameres and the intercalated discs of the sarcolemma, sarcoplasmic reticulum, mitochondria, T-tubules, and nuclei. Therefore, it is an interesting possibility that the differential splice pathways within the linker region of nebulin modify the affinity of nebulin's interaction with desmin. The specific interactions of nebulin and desmin were confirmed in vivo by yeast two hybrid experiments. To verify in the cellular level the interaction between nebulin isoform and desmin, we transfected COS-7 cell with EGFP-tagged nebulin and DsRed-tagged desmin. Based on evidence showing that despite exon 159 was deleted, the new isoform of nebulin was interact with desmin. This suggest that nebulin in brain may interact with another intermediate filament. The conservation of these ligand-binding capacity in brain and skeletal nebulins suggest that nebulins may have conserved roles in brain and skeletal muscle.

• Korean Journal of Poultry Science
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• v.30 no.3
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• pp.203-209
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• 2003

This experiment was conducted to investigate the influence of dietary supplementation of two different types of multiple probiotics in broiler chicks. Four hundred one day old male broiler chicks(Ross ${\times}$ Ross) were raised in the floor pen with five treatments(0, A: 0.10, 0.20%, B: 0.10, 0.20% of probiotics), consisting of 5 replicates for 5 weeks. Weight gain, feed intake and feed conversion were measured weekly, Total Lactobacillus, yeast, E. coli, Salmonella were examined from ileum and cecum at the end of experiment. Sera protein, cholesterol and ND vaccine titer were measured and fecal COD, CO$_2$ and NH$_3$ were detected in 24 hours after collection. Weight gain of chickes fed both types of probiotics increased for the first three weeks and showed significantly higher for the rest two weeks. Feed intake tended to be high in probiotics treatments compared with contro(P<0.05)1. Feed conversion improved significantly in B types of 0.2% probiotics compared with control. Total number of Salmonella was not consistent in ileum among treatments, whereas E. coli tended to be decreased in both types of probiotics compared with control. NH$_3$ and CO$_2$ emission of feces were decreased in both types of probiotics, but it was not statistically different(p>0.05). COD of chicks fed both probiotics was lower than control. There were no significantly different breast meat(P>0.05). Total cholesterol decreased in probiotics groups regardless of it's type. ND antibody vaccine titer was prone to be a small increments.

• Applied Biological Chemistry
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• v.48 no.3
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• pp.201-206
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• 2005

The development of microbial inoculant was conducted using a by-product of oriental herbal medicine. The constituent of the by-product, which was high in organic matter, was 11.3% of crude protein, 5.1% of crude lipid, 49.7% of NDF (neutral detergent fiber), and 33.8% of ADF (acid detergent fiber). Microorganisms isolated from the by-product of oriental herbal medicine were 35 species. Among them, 6 bacterial species, 4 fungal species, 2 actnomycetes species, and 1 yeast species were effective in the utilization of the by-products. The 13 strains screened were tested for the plant growth-promoting effect in soybean seedling. BL-333 strain was found to increase the soybean yield by about 23% as compared with control. The strain BL-333 was identified as Paenibacillus marcerans. P. marcerans BL-333 showed high anti-fungal activities against virulent fungi, especially Fusarium sp. and Collectotrichum sp. Yields of plants which were inoculated with microbial inoculant prepared with P. marcerans BL-333 and by-product of oriental herbal medicine were found to be higher than control by $3{\sim}24%$. The yield was especially promoted in lettuce, radish, chinese cabbage and cucumber plants.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.355-360
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• 2009

In this study, the arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was expressed on the cell surface of S. cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The constructed plasmid, pCTAST (7.1 kb), was introduced to S. cerevisiae EBY100 cell, and yeast transformants on YPDG plate showed the hydrolyzing activity for 4-methylumbelliferyl-sulfate and p-nitrophenyl-sulfate. When S. cerevisiae EBY100/pCTAST was grown on YPDG medium, the arylsulfatase activity of cell pellet reached about 1.2 unit/mL, whereas no extracellular arylsulfatase activity was detected. The DNA ladder in agarose prepared from agar by this recombinant arylsulfatase showed similar resolution and migration patterns with a commercial agarose. This results revealed that arylsulfatase expressed on the cell surface of S. cerevisiae could be applicable to the economic production of electrophoretic-grade agarose.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.53-58
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• 1983

Sik-hae is a traditional Korean fermented fish product which is made from flat-fish, garlic, salt, red pepper and millet. The changes in chemical composition, pH, acidity and the contents of Amino-N, VBN, TMA and organic acids were investigated during the fermentation of Sik-hae. The changes in the microflora, enzyme activity and the sensory quality including textural characteristics were also evaluated. The changes in the contents of crude protein, crude fat and moisture during Sik-hae fermentation were negligible. The pH of the product tended to decrease in the course of fermentation and it showed the minimum value of 4.5 after 7 days of fermentation. On the other hand the acidity continued to increase up to 2300mg % by 4 weeks of fermentation. Lactic acid was the major organic acid. The content of Amino-N in sik-hae gradually increased up to 673.6mg % by 2 weeks of fermentation, and then slightly decreased. The content of VBN increased rapidly during the first 2 weeks of fermentation, while little changes in TMA content was observed. The number of proteolytic bacteria increased slightly for the first 2 weeks and then rapidly decreased. The number of yeast and acid forming bacteria increased rapidly from the 4th day to the 14th day of fermentation and then decreased. Both lipase and protease activities showed the maximum at the 11th day of fermentation. The texture softening of the fish occurred after 1 week of fermentation and the adhesiveness appeared after 2 weeks of fermentation. Summarizing these results, the optimum fermentation time for Sik-hae from flat fish were 2 weeks at 2$0^{\circ}C$ and the quality of the product could be kept for up to 4 weeks in refrigerator.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.3
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• pp.401-411
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• 2015

Bitter melon (Momordica charantia L.) has various biological functions, including anti-diabetic, anti-cancer, anti-inflammatory, anti-viral, and antioxidant activities. However, the development of foods using bitter melon (BM) is unexplored due to its bitter taste. In this study, BM pickle was prepared, and changes in quality characteristics and antioxidant activity during ageing were determined. After 4 weeks of ageing, pH levels, salinities, soluble solids, and reducing sugars of BM pickle decreased, whereas acidities increased. In addition, total viable, lactic acid bacteria, and yeast viable cell numbers greatly increased until 1 week, after which they slightly decreased. Levels of soluble phenolics increased during ageing, antioxidant activity increased accordingly. Crude protein, K, and P contents of pickles were lower than in raw material of BM, whereas crude fat and Na contents increased. Corn silk extracts were added to BM pickle during ageing in order to improve the bitter taste and flavor. The best sensory quality was obtained by adding 0.25% corn silk extracts.

• Korean Journal of Food Science and Technology
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• v.38 no.4
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• pp.543-547
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• 2006

In order to improve wine quality, the selection of yeast strain and of additives in the manufacture of Bokbunja (Rubus coreanus Miq.) wine was investigated. The chemical composition of the edible portions of Bokbunja fruits was 86.5% moisture, 0.2% crude protein, 0.9% crude fat, 6.6% crude fiber, 0.5% ash and $10^{\circ}Brix$ sugar, and was 2.99% fructose, 2.53% glucose and 0.07% sucrose in fruit extract. The predominant organic acids in the fruit were citric acid (14.57 mg/mL) and malic acid (2.24 mg/mL) with smaller amounts of shikimic, pyroglutamic and oxalic acid. During fermentation, citric and malic acid levels decreased, while formic and acetic acid were released. Saccharomyces cerevisiae KCCM 12224 (Sc-24) was more favorable for alcoholic fermentation of Bokbunja and the addition of 200 ppm of potassium metabisulphite to must was more efficient than other $SO_2$ sources with a higher overall acceptability score. Sc-24 increased alcohol production from 9.8 to 14.8% in a sugar concentration dependent manner $(18-28^{\circ}Brix)$. The color value of early stage Bokbunja must was improved by supplementing with Japanese apricot extract, but this did not influence the color value of Bokbunja wine after primary fermentation. The astringent taste of Bokbunja wine was reduced by removing the seed from the fruit. Sugar solution (50%, w/v) was used instead of sugar power to prevent the possibility of undissolved sugar due to insufficient mixing. This substitution did not influence sensory evaluation.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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• pp.1287-1293
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• 2016

The objectives of this experiment was to evaluate the subsequent growth and organ weights, blood profiles and cecal microbiota of broiler chicks fed pre-starter diets containing fermented soybean meal products during early phase. A total of nine hundred 1-d-old chicks were randomly assigned into six groups with six replicates of 25 chicks each. The chicks were fed control pre-starter diet with dehulled soybean meal (SBM) or one of five experimental diets containing fermented SBM products (Bacillus fermented SBM [BF-SBM], yeast by product and Bacillus fermented SBM [YBF-SBM]; Lactobacillus fermented SBM 1 [LF-SBM 1]; Lactobacillus fermented SBM 2 [LF-SBM 2]) or soy protein concentrate (SPC) for 7 d after hatching, followed by 4 wk feeding of commercial diets without fermented SBMs or SPC. The fermented SBMs and SPC were substituted at the expense of dehulled SBM at 3% level on fresh weight basis. The body weight (BW) during the starter period was not affected by dietary treatments, but BW at 14 d onwards was significantly higher (p<0.05) in chicks that had been fed BF-SBM and YBF-SBM during the early phase compared with the control group. The feed intake during grower and finisher phases was not affected (p>0.05) by dietary treatments. During total rearing period, the daily weight gains in six groups were 52.0 (control), 57.7 (BF-SBM), 58.5 (YBF-SBM), 52.0 (LF-SBM 1), 56.7 (LF-SBM 2), and 53.3 g/d (SPC), respectively. The daily weight gain in chicks fed diet containing BF-SBM, YBF-SBM, and LF-SBM 2 were significantly higher values (p<0.001) than that of the control group. Chicks fed BF-SBM, YBF-SBM, and LF-SBM 2 had significantly lower (p<0.01) feed conversion ratio compared with the control group. There were no significant differences in the relative weight of various organs and blood profiles among groups. Cecal microbiota was altered by dietary treatments. At 35 d, chicks fed on the pre-starter diets containing BF-SBM and YBF-SBM had significantly increased (p<0.001) lactic acid bacteria, but lowered Coli-form bacteria in cecal contents compared with those fed the control diet. The number of Bacillus spp. was higher (p<0.001) in all groups except for LF-SBM 1 compared with control diet-fed chicks. At 7 d, jejunal villi were significantly lengthened (p<0.001) in chicks fed the fermented SBMs vs control diet. Collectively, the results indicate that feeding of fermented SBMs during early phase are beneficial to the subsequent growth performance in broiler chicks. BF-SBM and YBF-SBM showed superior overall growth performance as compared with unfermented SBM and SPC.

• Proceedings of the Microbiological Society of Korea Conference
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• 2007.05a
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• pp.120-122
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• 2007

All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.