• Korean Journal of Food Science and Technology
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• v.43 no.5
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• pp.553-557
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• 2011

Garlic specific organic compounds were separated and purified using a recycling preparative high-performance liquid chromatography (HPLC) from blanched garlic cloves. Identification of the compounds involved comparing the previously reported HPLC retention times as well as other identification methods including $^1H$- and $^{13}C$-nuclear magnetic resonance and liquid chromatography-mass spectrometry. The yields of garlic specific organic compounds were 12.2, 42.5, 1.6, 1.2, and 4.8% on wet weight basis of garlic for alliin(S-allyl-L-cysteine sulfoxide), isoalliin(S-1-propenyl-L-cysteine sulfoxide), ${\gamma}$-glutamyl-S-allylcysteine, ${\gamma}$-glutamyl-S-1-propenylcysteine and ${\gamma}$-glutamyl-phenylalanine, respectively. All the compounds, except for ${\gamma}$-glutamylphenylalanine, contained sulfur.

• The Korean Journal of Pesticide Science
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• v.10 no.1
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• pp.56-65
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• 2006

An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.

• Journal of Life Science
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• v.21 no.10
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• pp.1481-1486
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• 2011

Spent mushroom substrate (SMS) is a by-product remained after a crop of mushrooms. About seven thermophilic strains were isolated from SMS (Flammulina velvtipes). Among them, one isolate, designated UJ03, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ03 produced cellulase and xylanase as extracellular hydrolases. The strain UJ03 was identified as a member of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ03 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ03 was classified within the genus Bacillus, for which the name Bacillus sp. UJ03 is proposed. The antifungal compound from Bacillus sp. UJ03 was similar to lipopeptide iturin A of Bacillus sp.

• 한국유가공학회:학술대회논문집
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• 1996.11a
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• pp.18-29
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• 1996

1. CWPC중의 새로운 생리활성물질의 검색 Mouse 임파세포의 증식효과를 지표로 하는 면역기능을 검토하여 CWPC중의 면역 부활작용을 갖는 새로운 성분의 검색을 실시하였다. CWPC를 여러 가지 분획법으로 분획하여 mouse 임파세포의 증식효과를 지표로 면역 활성성분을 검색하였다. 그 결과 gel filtration, 음이온교환법을 사용하여 분획한 당을 다량 포함한 부분에 강한 면역부활담당세포에 대하여 증식활성을 나타내는 물질을 발견하였다. 이 물질은 SDS-PAGE상에서 분자량이 약 16kDa에 위치하여 Ca, P 및 당쇄를 포함한 물질이며, 이것을 GPP로 하였다. GPP에는 우유케이신의 trypsin분해물이며 Ca와 무기인을 풍부하게 포함하는 ${\beta}$-CPP와 유사한 phosphoserin 영역을 갖는 성분과 갖지 않는 성분의 2종류가 존재하며, 각각의 면역 부활활성이 인정되었다. 각 성분의 아미노산 분석, 당 분석의 결과에서 지금까지 보고된 우유중의 면역 담당세포에 대한 증식활성을 갖는 물질과는 상이한 성분인 것으로 밝혀졌다. 더욱이 이 활성물질 (GPP)은 PP cell에서도 동등한 활성이 있는 것으로 판단되었다. 이러한 결과를 종합하여 보면 CWPC중에서 지금까지 알려지지 않았던 새로운 면역 부활물질이 존재하며, 그 성분에는 CPP와 유사한 phophoserine 영역이 존재하는 성분이 포함되어 있고, N-글리코실 결합의 당쇄가 존재하는 것으로 시사되었다. 이 성분은 전신면역의 지표인 비장세포에 대해서만이 아니고, 장관면역계에 중요한 역할을 담당하는 PP cell에서도 활성이 있는 것으로 보아 전신 및 국부적인 면역기능의 부활성분으로서 응용의 가능성이 시사되었다. 2. GPP의 면역담당세포에 대한 증식활성의 메카니즘의 검토 CWPC중의 GPP의 면역담당세포증식활성의 메카니즘을 해명하기 위해 먼저 이 성분중의 어느 부분이 활성에 관여하는지를 pronase 분해 및 phophoserine 영역을 인식하는 항체를 사용하여 검토하였다. 그 결과 pronase 분해처리에서도 활성의 감소를 나타내지 않았으므로 이러한 활성에는 당이 필수 불가결하다는 점이 시사되었다. 또한 phosphoserine 영역을 인식하는 항체에 의해서도 활성은 감소하지 않는 것으로 보아 phosphoserine 영역이 세포증식활성에 관여하지 않는 것으로 판단되었다. 또한 분획한 면역담당세포에 대한 증식활성을 측정하는 것으로 이 성분의 표적면역담당세포를 동정하여, B세포에 대해서만 특이적으로 증식활성을 나타내는 것으로 밝혀졌다.

• Journal of Dairy Science and Biotechnology
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• v.31 no.2
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• pp.153-159
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• 2013

Lactic acid bacteria are microorganisms that are closely associated with human and/or animal environments, and are categorized as generally recognized as safe (GRAS) organisms due to their ubiquitous appearance in foods and their contribution to the healthy microflora of mucosal surfaces. This study was performed to isolate and identify lactic acid bacteria with antagonistic effects against food-borne pathogens. A total of 3,000 acid-producing bacteria were isolated from infant feces, cattle feces, goat feces, dog feces, pig feces, vaginal tracts, vegetables, fruits, Kimchi, Jeotgal, fermented sausages, raw milk, cheese, yogurt, Cheonggukjang, Meju, and Makgeolli cultured on MRS agar with 0.05% bromocresol purple. For the isolation of bacteriocin-producing bacteria, the diameter of the clear zone was measured on MRS agar plates. Twenty-six isolates exhibited strong antibacterial activity against indicator strains such as Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis. Lactic acid bacteria were identified as Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus curvatus, Lactobacillus plantarum, and Pediococcus acidilactici by 16S rDNA gene sequence analysis. The results of this study suggest that the isolates could be used as potential probiotic starters for functional food applications.

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.262-268
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• 2005

Intoxication of murine macrophages (RAW 264.7) with the anthrax lethal toxin (LeTx 100 ng/ml) results in profound alterations in the host cell gene expression. The role of LeTx in mediating these effects is unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, we have used two-dimensional polyacrylamide gel electrophoresis to analyze the protein profile of murine macrophages treated with the LeTx, and have coupled this to protein identification using MALDI-TOF mass spectrometry. Interpretation of the peptide mass fingerprint data has relied primarily on the ProFound database. Among the differentially expressed spots, cleaved mitogen-activated protein kinase kinase (Mek1) and glucose-6-phosphate dehydrogenase were increased in the LeTx treated macrophages. Mek1 acts as a negative element in the signal transduction pathway, and G6PD plays the role for the protection of the cells from the hyper-production of active oxygen. Our results suggest that this proteomic approach is a useful tool to study protein expression in intoxicated macrophages and will contribute to the identification of a putative substrate for LeTx.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.63-67
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• 2010

Proteases catalyze hydrolytic cleavage of a peptide bond between amino acids and occupy pivotal positions in application in physiological and commercial fields. During the screening for novel bacteria producing extracellular protease, two bacterial strains, WD12 and WD32, were isolated from rotten trees and they made clear zone on LB plates supplemented with 1% skim milk. The similarities of 16S rRNA gene sequence of either WD12 or WD32 to GenBank database showed the highest to Pseuoxanthomonas mexicana as 97.8 and 99.8%, respectively. Phylogenetic analysis showed that both isolated was located within the cluster comprising P. mexicana and P. japonesis. WD12 and WD32 were catalase- and oxidase-positive, Gram-negative rod strains. In case of WD12, it could assimilate malate, but could not assimilate D-mannose, which were different characteristics from P. mexicana. Both Pseuoxanthomonas sp. WD12 and WD32 optimally produced extracellular protease at $35-37^{\circ}C$, and maximal activity showed as 656 unit/ml and 267 unit/ml, respectively.

• Journal of Life Science
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• v.20 no.2
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• pp.269-274
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• 2010

An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.3
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• pp.266-270
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• 1999

Two forms of gonadotropin releasing hormone (GnRH) are identified in the brain of adult mature spotted sea bass (Lateolabrax sp.) by immunohistochemical methods. Salmon GnRH immunoreactive (sGnRH-ir) cell bodies were distributed in the olfactory bulb, ventral telencephalon and preoptic region. Immunoreactive fibers were observed in the vicinity of the brain including the olfactory bulbs, the telencephalon, the optic nerve, the optic tectum, the cerebellum, the medulla oblongata and rostral spinal cord. In most cases, these fibers did not form well defined bundles. However, there was a clear continuum of immunoreactive fibers, extending from the olfactory bulbs to the pituitary. cGnRH-II-ir cell bodies were only found in olfactory bulbs. However, the distribution of cGnRH-II-ir fibers was basically similar to that of sGnRH-ir fibers except for the absence of their continuity between the olfactory bulbs and the pituitary. These data suggest that sGnRH and cGnRH-II are endogenous peptides and indicate the presence of multiple neuroendocrine functions in the brain of the spotted sea bass. It seems that sGnRH not only regulates GTH secretion but also functions as a neurotransmitter, whereas cGnRH-II functions only as a neurotransmitter.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.547-554
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• 2010

Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.