• Title/Summary/Keyword: yeast cell wall

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Evaluation of Six Species Ciliates as a Live Food and Culture Environment for Euplotes sp. (먹이생물로써의 섬모충 6종의 평가와 Euplotes sp.의 배양 환경)

  • Yoo Jin Hyung;Hur Sung Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.4
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    • pp.342-347
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    • 2002
  • Ciliates have the possibility of a new live food in marine finfish culture because of their wide range of body size, thin tell wall, show motility, and fast reproduction rate. In this research, six species of ciliates were isolated from south coast and salt pond in Korea. The fitness of these species as a live food was evaluated in terms of size, motility, suspensibility and cell density. As the result, Euplotes sp. (K-1) was found suitable to be a new live food which might substitute rotifers, Brachionus plintilis and B. rotundiformis in fish larvae culture. The modified $F{\emptyset}yn's$ Erdschreiberd media, MErds-2 with the addition of glycine, glucose and yeast extract increased six times higher growth rate of Euplotes sp. (K-1) than the basic F$\emptyset$yn's Erdschreiberd media. The optimum water temperature, pH and light intensity for this ciliates were $22.5^{\circ}C$, 8 and 2,000 lux, respectively, and its culture environmental range was relatively wide, On the other hand, this ciliate fed baker's yeast, Saccharomyces cererisiae grew up to 1,240 inds./mL with the inocula of 100 inds./mL within 7 days. The results of the study showed that Euplotes sp. (K-1) has a potential to be utilized as a new live food in fish larvae culture.

Characterization of Antibacterial Substance - Producing Bacillus subtilis Isolated from Traditional Doenjang (전통 된장으로부터 분리한 향균물질 생산 Bacillus subtilis의 특성)

  • Ryu, Hyun-Soon;Shon, Mi-Yae;Cho, Soo-Jeong;Park, Seok-Kyu;Lee, Sang-Won
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.87-94
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    • 2007
  • A bacterium which has high enzymatic activities such as amylase, cellulase and protease was isolated from Korean traditional soybean food, doenjang. The isolated bacterium was identified to Bacillus subtilis HS25 by the test of morphological and biochemical properties according to Bergey's Manual of Systematic Bacteriology and API 50 CHL kit, and by the 16S rDNA sequence. The isolated B. subtilis HS25 had a potent antibacterial activity against food born causative or pathogenic bacteria. B. subtilis HS25 is endospore forming cell and contained flagella and abundant viscous material at the out layer of cell wall. It was rod type bacterium $(0.5{\sim}0.8{\times}3{\sim}5{\mu}m)$ having biochemical characteristics such as gram staining(+), catalase(+), oxidase(-) and hydrolysis of esculin(+). The optimal medium compositions for production of antibacterial substance in the B. subtilis HS25 were 1% of soluble starch, 0.5% of yeast extract, 0.5% of peptone and 0.05% of MgCl$_2{\cdot}6H_{2}O$. The optimum temperature and pH of the growth of the B. subtilis HS25 was 35$^{\circ}C$ and pH 7.5, respectively. The antibacterial activity was more high in neutral to a little alkaline pH (6.5-10.5) than in acidic pH. The optimal shaking speed to grow and to produce antibacterial substance of the B. subtilis HS25 was 160${\sim}$200 rpm. The optimal culture time for antibacterial activities of the bacterium were shown to be in the range of 12-36 hr.

Functional Characterization and Application of the HpOCH2 Gene, Encoding an Initiating $\alpha$l,6-Mannosyltransferase, for N-glycan Engineering in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Moo-Woong;Kim, Eun-Jung;Kim, Jeong-Yoon;Rhee, Sang-Ki;Kang, Hyun-Ah
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.278-281
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    • 2004
  • The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

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Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Changes of Volatile Flavor Compounds of Seibel Grape Must during Alcohol Fermentation and Aging (Seibel 포도즙 알코올 발효 및 저장 중 휘발성 향기성분의 변화)

  • 고경희;장우영
    • Microbiology and Biotechnology Letters
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    • v.27 no.6
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    • pp.491-499
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    • 1999
  • A great variety of the volatile metabolic by-products was formed in yeast cell during alcohol fermentation. The seibel grape (Vitis labrasca) which was grown in the Southern Korea used for wines. The objective of this research was to identify the volatile flavor compounds during alcohol fermentation and aging at 12$^{\circ}C$. saccharomyces cerevisiae and Schizosaccharomyces pombe were inoculated and fermented in seibel grape must. The volatile flavor compounds of logarithmic, stationary and death phases were extracted, concentrated and identified by gas chromatography/mass spectrometer (GC/MS). The volatile flavor compounds were determined by a Hewlett-Packard 5890 II Plus GC which was equipped with Supelcowax 10 fused silica capillary column (60m$\times$0.32mm$\times$0.25${\mu}{\textrm}{m}$ film thickness) wall coated with polyethyleneglycerol. The scan detection method allowed the comparison of the spectrum from the chromatogram of volatile flavor compounds to those in data Wileynbs base library. Among the volatile compounds collected by ether-hexane extraction method, the evolution of 20 main compounds, such as 9 esters (ethyl butyrate, isoamyl acetate, ethyl caproate, n-hexyl acetate, ethl caprylate, ethyl caprate, diethy succinate, ethyl hexadecanoate, 2-pheneethyl acetate), 4 alcohols (3-methyl-1-butanol, 1-hexanol, 1-heptanol, benzoethanol), 4 ketones and acids (2-octanone, caproic acid, caprylic acid, capric acid), 2 furan and phenol (2,6-bis(1,1-dimethyl ethyl)phenol, 2,3-dihydrobenzofuran) were observed during alcohol fermentation and aging. The production of the esters during alcohol fermentation with S. cerevisiae was higher than those of Sch. pombe. The sensory scores of the aged wine samples in aroma, taste and overall acceptability were not significantly different(p<0.05).

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Characterization of Acetobacter pomorum KJY8 Isolated from Korean Traditional Vinegar

  • Baek, Chang Ho;Park, Eun-Hee;Baek, Seong Yeol;Jeong, Seok-Tae;Kim, Myoung-Dong;Kwon, Joong-Ho;Jeong, Yong-Jin;Yeo, Soo-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1679-1684
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    • 2014
  • Acetobacter sp. strains were isolated from traditional vinegar collected in Daegu city and Gyeongbuk province. The strain KJY8 showing a high acetic acid productivity was isolated and characterized by phenotypic, chemotaxonomic, and phylogenetic inference based on 16S rRNA sequence analysis. The chemotaxonomic and phylogenetic analyses revealed the isolate to be a strain of Acetobacter pomorum. The isolate showed a G+C content of 60.8 mol%. It contained $\small{LL}$-diaminopimelic acid ($\small{LL}$-$A_2pm$) as the cell wall amino acid and ubiquinone $Q_9$ (H6) as the major quinone. The predominant cellular fatty acids were $C_{18:1}w9c$, w12t, and w7c. Strain KJY8 grew rapidly on glucose-yeast extract (GYC) agar and formed pale white colonies with smooth to rough surfaces. The optimum cultivation conditions for acetic acid production by the KJY8 strain were $20^{\circ}C$ and pH 3.0, with an initial ethanol concentration of 9% (w/v) to produce an acetic acid concentration of 8% (w/v).

Antimicrobial Activity of the Synthetic Peptide Scolopendrasin II from the Centipede Scolopendra subspinipes mutilans

  • Kwon, Young-Nam;Lee, Joon Ha;Kim, In-Woo;Kim, Sang-Hee;Yun, Eun-Young;Nam, Sung-Hee;Ahn, Mi-Young;Jeong, MiHye;Kang, Dong-Chul;Lee, In Hee;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1381-1385
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    • 2013
  • The centipede Scolopendra subpinipes mutilans is a medicinally important arthropod species. However, its transcriptome is not currently available and transcriptome analysis would be useful in providing insight into a molecular level approach. Hence, we performed de novo RNA sequencing of S. subpinipes mutilans using next-generation sequencing. We generated a novel peptide (scolopendrasin II) based on a SVM algorithm, and biochemically evaluated the in vitro antimicrobial activity of scolopendrasin II against various microbes. Scolopendrasin II showed antibacterial activities against gram-positive and -negative bacterial strains, including the yeast Candida albicans and antibiotic-resistant gram-negative bacteria, as determined by a radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin II bound to the surface of bacteria through a specific interaction with lipoteichoic acid and a lipopolysaccharide, which was one of the bacterial cell-wall components. In conclusion, our results suggest that scolopendrasin II may be useful for developing peptide antibiotics.

Comparative Study of Immune-Enhancing Activity of Crude and Mannoprotein-Free Yeast-Gluean Preparations

  • Kim, Hye-Nam;Lee, Jung-Nam;Kim, Gi-Eun;Ha-Lee, Young-Mie;Kim, Chan-Wha;Sohn, Jeong-Won
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.826-831
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    • 1999
  • ${\beta}-Glucan$, one of the major cell wall components of Saccharomyces cerevisiae, is known to enhance the immune function, especially by activating macrophages. Accordingly, in an effort to develop a safe and efficient immune stimulatory agent, we prepared crude ${\beta}-glucan$ (glucan-p1) and partially purified ${\beta}-glucan$ that was free of mannoproteins (glucan-p2), and evaluated their effect on both the macrophage function and resistance to E. coli-induced peritonitis. To investigate the function of the macrophages, phagocytosis, $TNF-{\alpha}$ secretion, oxygen burst, and the expression of cytokine genes such as $IFN-{\gamma}$ and IL-12 were analyzed. Glucan-p2 markedly stimulated the macrophages with all these parameters. Glucan-p1, however, did not stimulate phagocytosis, yet it induced $TNF-{\alpha}$ secretion, oxygen burst, and the expression of $IFN-{\gamma}$ and IL-12, although less efficiently than glucan-p2. Finally, to test the in vivo protective effect of {\beta}-glucan against infection, the survival of mice from E. coli-induced peritonitis was investigated. After 24 h of the peritoneal challenge of E. coli, all of the mice treated with glucan-p2 survived whereas none survived in the control group. Glucan-p1 showed only a marginal effect in protecting the mice. These results suggest that mannoprotein-free gluean-p2, but not gluean-p1, can serve as an effective immune-stimulating agent.

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Inhibition Activity Against Pathogenic Organism of Probiotic Bacteria and Characterization of Inhibition Activity of Isolated Bacteria from Calf Dejecta (Probiotic균주의 Pathogenic Organism에 대한 억제 활성과 송아지분변 분리균주의 억제활성 특성)

  • 배임희;변정열;배귀석;이상석;장문백;윤영호
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.907-920
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    • 2006
  • This study was conducted to investigate the inhibitory activity of Lactobacillus spp., Bacillus ssp., and calf fecal isolates against pathogenic Salmonella typhimurium, E. coli, Listeria monocytogenes, and Staphylococcus aureus. Among thirteen strains of Lactobacillus ssp. tested, Lactobacillus helveticus CU631 showed the highest inhibition against three pathogens, whereas Bacillus spp. showed a weak inhibitory activity. Four calf fecal isolates were identified as Lactobacillus pentosus CU13, CU05, Pediococcus pentosaceus CUR02, and Lactobacillus lactis ssp. lactis CUM14. The whole cell and cell wall components of L. rhamnosus CU02 and L. pentosus CU13 were active in the inhibition of L. monocytogenes. The medium components and levels, which affect on the inhibitory activity, were revealed as Tween 80 1.0%, peptone 3.0%, yeast extract 3.0%, glucose 3.0%, beef extract 3.0%, and NaCl 1.0~3.0%, respectively. Inhibitory activity of the supernatant culture medium was not affected by catalase and proteinase K treatment but affected by heat treatment at 80℃ and netralization, which implies that the inhibitory activity is due to the production of organic acids during the growth. L. pentosus CU13 and L. rhamnosus CU02 exhibited broad inhibition spectrum against 16 out of 21 strains including some pathogens. Oral administration of L. rhamnosus CU02 to the mice infected with E. coli O157:H7 was proven to be effective to recover their body weight during the experimental period.

Studies on the Enzyme from Arthrobacter luteus Accelerating the Lysis of Yeast Cell Walls -II. Separation of the Factor Accelerating the Lysis of Yeast Cell Walls from the Preparation of Crude Zymolyase and Partial Purification of the Zymolyase with the Sephadex G-75 Gel- (Arthrobacter luteus가 생산(生産)하는 효모세포벽(酵母細胞壁) 용해촉진효소(溶解促進酵素)에 관(關)한 연구(硏究) -제 2 보(第2報) : Crude Zymolyase 표품중(標品中)으로부터 효모(酵母) 세포벽(細胞壁) 용해(溶解) 촉진(促進) 인자(因子)의 분리(分離) 및 Sephadex G-75 Gel에 의한 Zymolyase의 부분(部分) 정제(精製)-)

  • Oh, Hong-Rock;Shimoda, Tadahisa;Funatsu, Masaru
    • Korean Journal of Food Science and Technology
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    • v.12 no.4
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    • pp.254-262
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    • 1980
  • A series of experiment were carried out to separate the factor accelerating the lysis of cell wall of $Saccharomyces\;sak{\acute{e}}$ from the preparation of crude zymolyase obtained from Arthrobacter luteus. An attempt was also made to purify the enzyme which is essential for the study on the separation of the factor. The results are summarized as follows: 1. Crude zymolyase was fractionated 5 peaks $(A{\sim}E)$ containing three peaks $(A{\sim}C)$ passed through the column by the chromatography on Biogel CM-30. 2. Among the five peaks, peak E (protease fraction) was found to contain the factor accelerating the lytic activity of the zymolyase. 3. L-c fraction purified in almost free form from the nonlytic ${\beta}-1$, 3-glucanase, protease and inert protein by the affinity adsorption chromatography with Sephadex G-75 gel was obtained from zymolyase fraction (peak D). When it was subjected to polyacrylamide gel disc electrophoresis, only one clear protein band was observed at pH 4. 5, but still detected two or more band at pH 8. 3.

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