• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.452-460
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• 2016

Acid stress can affect the viability of probiotics, especially Bifidobacterium. This study aimed to improve the acid tolerance of Bifidobacterium longum BBMN68 using adaptive evolution. The stress response, and genomic differences of the parental strain and the variant strain were compared by acid stress. The highest acid-resistant mutant strain (BBMN68m) was isolated from more than 100 asexual lines, which were adaptive to the acid stress for 10^th, 20^th, 30^th, 40^th, and 50^th repeats, respectively. The variant strain showed a significant increase in acid tolerance under conditions of pH 2.5 for 2 h (from 7.92 to 4.44 log CFU/ml) compared with the wild-type strain (WT, from 7.87 to 0 log CFU/ml). The surface of the variant strain was also smoother. Comparative whole-genome analysis showed that the galactosyl transferase D gene (cpsD, bbmn68_1012), a key gene involved in exopolysaccharide (EPS) synthesis, was altered by two nucleotides in the mutant, causing alteration in amino acids, pI (from 8.94 to 9.19), and predicted protein structure. Meanwhile, cpsD expression and EPS production were also reduced in the variant strain (p < 0.05) compared with WT, and the exogenous WT-EPS in the variant strain reduced its acid-resistant ability. These results suggested EPS was related to acid responses of BBMN68.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.139-139
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• 2017

The complete chloroplast genome sequence of Avena sterilis L., a dominant wild oat species in the family Poaceae, is first reported in this study. The complete cp genome sequence of A. sterilis is 135,887 bp in length with 38.5% overall GC content and exhibits a typical quadripartite structure comprising one pair of inverted repeats (21, 603 bp) separated by a small single-copy region (12,575 bp) and a large single-copy region (80,106). The A. sterilis cp genome encodes 111 unique genes, 76 of which are protein-coding genes, 4 rRNA genes, 30 tRNA genes and 18 duplicated genes in the inverted repeat region. Nine genes contain one or two introns. Pair-wise alignments of cp genome were performed for genome-wide comparison. This newly determined cp genome sequence of A. sterilis will provide valuable information for the future breeding programs of valuable cereal crops in the family Poaceae.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.230-230
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• 2002

Class Ⅲ chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class Ⅲ chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5′flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• Proceedings of the Korea Information Processing Society Conference
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• 2004.11a
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• pp.805-808
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• 2004

본 논문에서는 두 단백질의 구조적 유사성을 기반으로 한 단백질 비교를 위해서 전처리 기법으로서의 주성분분석기법을 소개한다. 기존의 백본 및 알파탄소 간의 거리행렬(distance matrix), 2차 구조 비교기법, 구역(segment)단위의 비교 기법과 같은 단백질 비교 기법들은 위치이동(translation)와 회전(rotation)에 불변한(invariant) 차이를 구하기 위하여 거리행렬을 이용하였다. 그리고, 난 다음 이들의 최적화 과정을 거쳤다. 그러나, 본 논문에서 제시하는 전처리 기법으로서의 주성분분석기법은 단백질 구조를 전체적인 구조 관점에서 위치를 정렬시킨 후에 단백질 간의 구조를 비교하는 방식이다. 단백질의 구조의 방향성(Orientation)을 맞춘 다음에는 다양한 단백질 표현으로 구를 비교할 수 있다. 본 논문에서는 두 단백질의 구조의 유사성을 측정하기 위한 간결한 단백질 표현(representation)으로 3 차원 에지 히스토그램을 사용하였다. 이 기법은 방향성을 정렬하기 위하여 기존의 방법에서 사용되었던 반복적인 거리계산을 통한 최적화하는 과정을 없앰으로써 단백질 구조 비교 시간을 단축할 수 있는 새로운 단백질 구조 비교 패러다임을 가능하게 한다. 따라서, 이 패러다임을 통하여 적절한 단백질 구조 방향성 정렬과 단백질 구조 표현을 이용한 단백질 구조 비교 검색 시스템은 많은 양의 단백질 구조 정보로부터 원하는 형태의 단백질 구조를 빠른 시간에 검색할 수 있는 장점을 가질 수 있다.

• Food Engineering Progress
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• v.14 no.2
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• pp.118-126
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• 2010

The fibrous material fraction as a by-product from the commercial aloe vera gel processing was obtained and freeze dried. The physicochemical characteristics such as the proximate composition, crystalline/surface structures and several physical functionalities including the water holding capacity (WHC), swelling capacity (SW), oil holding capacity (OHC), emulsion/foam properties and viscosity properties of this powdered sample (100 mesh) were investigated and analyzed by comparison with commercial $\alpha$-cellulose as a reference sample. The total dietary fiber content of powdered sample was very high as much as 87.5%, and the insoluble dietary and soluble dietary fiber content ratios were 77.6 and 22.4%, respectively. The FT-IR spectrum of powdered sample showed a typical polysaccharide property and exhibited a x-ray diffraction pattern for cellulose III and IV like structure. SW (8.24${\pm}$0.15 mL/g), WHC(6.40${\pm}$0.19 g water/g solid) and OHC(10.32${\pm}$0.29 g oil/g solid) of freeze dried aloe cellulose were about 3.3, 1.4 and 2 times higher than those of commercial $\alpha$-cellulose, respectively. Aloe cellulose (~2%, w/v) alone had no foam capacity while improved the foam stability of protein solution (1% albumin+0.5% $CaCl_{2}$) by factor of 300%. Emulsion capacity of 2%(w/v) aloe cellulose was about 70% level of 0.5%(w/v) xanthan gum, but its emulsion stability was about 1.2 times higher than that of xanthan gum. Also, aloe cellulose containing CMC (carboxyl methyl cellulose) of 0.3%(w/v) showed a very good dispersity. Aloe cellulose dispersion of above 1%(w/v) exhibited higher pseudoplasticity and concentration dependence than those of $\alpha$-cellulose dispersion, indicating the viscosity properties for new potential usage such as an excellent thickening agent.

• Animal Bioscience
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• v.34 no.1
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• pp.56-65
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• 2021

Objective: This study was conducted to investigate the effects of molasses and Lactobacillus plantarum on the ensiling quality and in vitro rumen fermentation of sudangrass silage prepared with or without wilting. Methods: The ensiling experiment, measured with 3 replicates, was carried out according to a 2×4 (wilted stages×additives) factorial treatment structure. Dry matter of the fresh (210 g/kg fresh matter) or wilted (305 g/kg fresh matter) sudangrass were ensiled (packed into 5.0-L plastic jars) without additive (control) or with molasses (M), Lactobacillus plantarum (LP), or molasses + Lactobacillus plantarum (M+LP). After 60 days of ensiling, the silages were analyzed for the chemical, fermentation, and in vitro characteristics. Results: After 60 days of ensiling, the fermentation parameters were affected by wilted, the additives and the interactions of wilted with the additives (p<0.05). The M+LP treatment at wilted had higher lactic acid levels and V-score (p<0.05) but lower pH values and butyric acid concentrations than the other treatments. In comparison with sudangrass before ensiling, after ensiling had lower dry matter and higher non-fibrous carbohydrate. The in vitro gas production, in vitro dry matter digestibility, in vitro crude protein digestibility, and in vitro acid fiber detergent digestibility changed under the effects of the additives. Significant interactions were observed between wilted and the additives in terms of in vitro gas production at 48 h, asymptotic gas production, gas production rate, half time, and the average gas production rate. The total volatile fatty acid levels in the additive treatments were higher than those in the control. Conclusion: Wilting and supplementation with molasses and Lactobacillus plantarum had the ability to improve the ensiling quality and in vitro nutrient digestibility of sudangrass silage. The M+LP treatment at wilted exhibited the strongest positive effects on silage quality and in vitro ruminal fermentation characteristics.

• Food Science of Animal Resources
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• v.43 no.1
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• pp.46-60
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• 2023

Slaughterhouse blood is a by-product of animal slaughter that can be a good source of animal protein. This research purposed to examine the functional qualities of the blood plasma from Hanwoo cattle, black goat, and their hydrolysates. Part of the plasma was hydrolyzed with proteolytic enzymes (Bacillus protease, papain, thermolysin, elastase, and α-chymotrypsin) to yield bioactive peptides under optimum conditions. The levels of hydrolysates were evaluated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis. The antioxidant, metal-chelating, and angiotensin I-converting enzyme (ACE) inhibitory properties of intact blood plasma and selected hydrolysates were investigated. Accordingly, two plasma hydrolysates by protease (pH 6.5/55℃/3 h) and thermolysin (pH 7.5/37℃/3-6 h) were selected for analysis of their functional properties. In the oil model system, only goat blood plasma had lower levels of thiobarbituric acid reactive substances than the control. The diphenyl picrylhydrazyl radical scavenging activity was higher in cattle and goat plasma than in proteolytic hydrolysates. Ironchelating activities increased after proteolytic degradation except for protease-treated cattle blood. Copper-chelating activity was excellent in all test samples except for the original bovine plasma. As for ACE inhibition, only non-hydrolyzed goat plasma and its hydrolysates by thermolysin showed ACE inhibitory activity (9.86±5.03% and 21.77±3.74%). In conclusion, goat plasma without hydrolyzation and its hydrolysates can be a good source of bioactive compounds with functional characteristics, whereas cattle plasma has a relatively low value. Further studies on the molecular structure of these compounds are needed with more suitable enzyme combinations.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.47-53
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• 2010

Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.76-81
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• 2012

Silk protein has been explored to be used for biomedical applications for several decades. However, it has not been used in this field cause to their irreversible crystallization after dissolving in water. The existing methods of silk protein hydrolysis using silkworm cocoon were used with harmful solvents and through a very complicated process. Therefore, we have developed novel methods for the production of water-soluble hydrolysate using silkworm gland. We manufactured two types of silkworm gland-derived hydrolysate (water-soluble SGH, SSGH; total SGH, TSGH) and compared the characteristics with commercial cocoon-derived sericin hydrolysate (CSH). The molecular weight of SGH ranged from 7 to 50 kDa (SSGH) and 5 to 15 kDa (TSGH) within glycine, alanine, and aspartic acid as a main amino acid composition. In contrast, CSH ranged from 15 to 50 kDa within serine and aspartic acid. The results of FTIR implied that SGH was more soluble form than CSH, as shown by the decrease in the ${\beta}$-sheet structure at $1630cm^{-1}$ on amide I peak. In comparison with 10% fetal bovine serum, 0.1% (1 mg/ml) SSGH had equivalent effect on the proliferation of human dermal fibroblasts and mesenchymal stem cells. All results of the SSGH made by novel manufacturing process indicate the SSGH is more preferable as a culture medium supplement than cocoon-derived sericin.

• Plant Biotechnology Reports
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• v.5 no.4
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• pp.331-344
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• 2011

The well-conserved NBS domain of resistance (R) genes cloned from many plants allows the use of a PCR-based approach to isolate resistance gene analogs (RGAs). In this study, we isolated an RGA (CapRGC) from Capsicum annuum "CM334" using a PCR-based approach. This sequence encodes a protein with very high similarity to Rx genes, the Potato Virus X (PVX) R genes from potato. An evolutionary analysis of the CapRGC gene and its homologs retrieved by an extensive search of a Solanaceae database provided evidence that Rx-like genes (eight ESTs or genes that show very high similarity to Rx) appear to have diverged from R1 [an NBS-LRR R gene against late blight (Phytophthora infestans) from potato]-like genes. Structural comparison of the NBS domains of all the homologs in Solanaceae revealed that one novel motif, 14, is specific to the Rx-like genes, and also indicated that several other novel motifs are characteristic of the R1-like genes. Our results suggest that Rx-like genes are ancient but conserved. Furthermore, the novel conserved motifs can provide a basis for biochemical structural. function analysis and be used for degenerate primer design for the isolation of Rx-like sequences in other plant species. Comparative mapping study revealed that the position of CapRGC is syntenic to the locations of Rx and its homolog genes in the potato and tomato, but cosegregation analysis showed that CapRGC may not be the R gene against PVX in pepper. Our results confirm previous observations that the specificity of R genes is not conserved, while the structure and function of R genes are conserved. It appears that CapRGC may function as a resistance gene to another pathogen, such as the nematode to which the structure of CapRGC is most similar.