• Title/Summary/Keyword: cell wall protein

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Dicyma sp. YCH-37이 생산하는 효모세포벽 용해효소 I. 생산균주의 분리 및 효소의 정제

  • Chung, Hee-Chul;Hahm, Byoung-Kwon;Bai, Dong-Hoon;Hasegawa, Toru;Yu, Ju-Hyun
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.445-451
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    • 1996
  • The strain YCH-37, which produces yeast cell wall lytlc enzyme, was isolated from soil. From the microscopic observation, morphological and cultural characteristics, this strain was identified to fungus, Dicyma sp. So, we named this strain as Dicyma sp. YCH-37. The lytic enzyme effectively lysed Salmonella typhimurium among intact living bacteria and Torulopsis, Hansenula, Zygosaccharomyces among intact living yeast, as well as autoclaved yeast strains. The yeast cell wall lytic enzyme was succesively purified to 204 folds with 13% yields through yeast glucan affinity adsorption and DEAE-cellulose column chromatography. The enzyme was identified to monomeric protein with molecular weight of 25,000 daltons from the results of SDS-PAGE and gel filtration. The optimum pH and temperature for the yeast lytic activity were 8.0 and 50$\circ$C, respectively. The enzyme was stable up to 40$\circ$C, and between pH 4.0-pH 10.0.

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Transformation of Cell Wall-weakened Perilla Seedlings Using Phenolic Compound-treated Agrobacterium Cells and Recombinant Protein Expression (페놀화합물 처리 Agrobacterium 및 세포벽 약화 들깨새싹을 이용한 형질전환과 재조합 단백질 발현)

  • Chung, Il-Kyung;Shin, Dong-Il;Park, Hee-Sung
    • KSBB Journal
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    • v.24 no.6
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    • pp.598-601
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    • 2009
  • Perilla [Perilla frutescens (L.) Britt] seedlings are easy to grow and eaten as the health vegetable sprout. Two day old perilla seedlings since germination were given a mild wounding using cell wall lytic NaOH/SDS solution for infiltration with recombinant Agrobacterium cells treated with phenolic compounds. In the analysis of fluorometric GUS gene expression for the transformed perilla seedlings, GUS enzyme activity was the highest by the combined treatments of 50 mM acetosyringone and 0.5% NaOH solution containing 0.01% SDS implying a synergic effect. This result could be successfully applied for demonstrating hepatitis B virus antigen (HBsAg) protein expression.

Selective Removal of Protease from Soybean Cell Wall Degrading Enzyme Complex Isolated from Aspergillus niger CF-34 (Aspergillus niger CF-34로부터 분리한 대두세포벽분해효소 복합체 중의 Protease의 선택적인 제거)

  • Choi, Yeon-Bae;Kim, Kang-Sung;Sohn, Heon-Soo
    • Korean Journal of Food Science and Technology
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    • v.27 no.3
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    • pp.370-374
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    • 1995
  • By exposing the complex enzyme solution to alkaline condition, it was possible to remove the protease activity selectively without inactivation of soybean cell wall degrading activity of the crude enzyme complex produced by Aspergillus niger CF-34. Optimum reaction conditions were as follow. pH was $9.0{\pm}0.1$, temperature was $20^{\circ}C$ and reaction time was 30 min with gentle stirring. Over 90% of protease activity could be eliminated while the activities of pectinase, polygalacturonase, xylanase, carboxymethyl cellulase and soybean cell wall degrading enzyme were maintained to $80{\sim}100%$. Through alkali treatment, it was discovered that the quality and organoleptic properties of soy protein produced by this enzymes were improved because the hydrolysis of protein and formation of bitter peptide were decreased.

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Functional Analysis of the Tomato Spotted Wilt Virus(TSWV) NSm Protein by Using Immunoblotting and Immunogold Labelling Assay

  • Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.468-473
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    • 1996
  • The genome of tomato spotted wilt virus (TSWV) is composed of three RNA segments, S, M, and L RNA and the 5.0 kb M RNA encodes two glycoproteins Gl, G2 and NSm protein of unknown function. In an effort to investigate the function of the NSm protein, antibody was raised against NSm fusion protein overexpressed in Escherichia coli. This antibody was used to detect the NSm protein by using western blot analysis and electron microscopic observation after immunogold labelling. For the cloning of the NSm gene, total RNA extracted from a TSWV infected plant was used for cDNA synthesis and polymerase chain reaction (PCR) instead of going through time-consuming virus purification. A protein band specifically reacting to the NSm antibody was detected from TSWV inoculated plants. The NSm protein was detected in the cell wall fraction and in pellet from low speed centrifugation when the infected plant tissue was fractionated into 4 fractions. In the immuno-electron microscopic observation, gold particles were found around the plasmodesmata of infected plant tissue. These results suggest that the NSm protein of TSWV plays some role in cell-to-cell movement of this virus.

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Phenotypic and Cell Wall Proteomic Characterization of a DDR48 Mutant Candida albicans Strain

  • El Khoury, Pamela;Salameh, Carell;Younes, Samer;Awad, Andy;Said, Yana;Khalaf, Roy A.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.11
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    • pp.1806-1816
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    • 2019
  • Candida albicans is an opportunistic fungus possessing multiple virulence factors controlling pathogenicity. Cell wall proteins are the most important among these factors, being the first elements contacting the host. Ddr48 is a cell wall protein consisting of 212 amino acids. A DDR48 haploinsufficient mutant strain was previously found necessary for proper oxidative stress response and drug resistance. In this study, we aimed to further elucidate the role of Ddr48 by performing additional phenotypic characterization assays. A combinatory proteomic and bioinformatics approach was also undertaken to determine differentially expressed cell wall proteins. Results showed that the mutant strain exhibited a 10% decrease in adhesion mirrored by a 20% decrease in biofilm formation, and slight sensitivity to menadione, diamide, and SDS. Both strains showed similar hyphae formation, virulence, temperature tolerance, and calcofluor white and Congo red sensitivities. Furthermore, a total of 8 and 10 proteins were identified exclusively in the wild-type strain grown under filamentous and non-filamentous conditions respectively. Results included proteins responsible for superoxide stress resistance (Sod4 and Sod6), adhesion (Als3, Hyr4, Pmt1, and Utr2), biofilm formation (Hsp90, Ece1, Rim9, Ipp1, and Pra1) and cell wall integrity (Utr2 and Pga4). The lack of detection of these proteins in the mutant strain correlates with the observed phenotypes.

The effect of some detergents on the changes of bacterial membrane (계면 활성제 처리에 의한 세균 세포막의 변화에 관한 연구)

  • 이종삼;이호용;조기승;조선희;장성열;최영길
    • Korean Journal of Microbiology
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    • v.21 no.3
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    • pp.115-126
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    • 1983
  • The results that the effect of 6 detergents on the structural changes and biochemical composition of bacterial membrane of Escherichia coli and Bacillus cereus are as follows ; 1. Population growth of the bacteria was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was decreased by sodium dodecyl sulfate and palmitoyl choline, in E.coli and was decreased by palmitoyl carnitine and palmitoyl choline at the low concentration, in B. cereus. 2. The electron micrograph showed that cell wall lysis or cell collapse were observed in the treatment of sodium dodecyl sulfate and palmitoyl choline, and also cell wall was condensed by triton X-100 and sodium deoxy cholate, in E.coli. And in B. cereus, endospore formation of the bacteria was stimulated by palmitoyl choline, and cell lysis or structural changes of the membrane were observed in the treatment of sodium dodecyl sulfate, sodium cholate, and triton X-100, respectively. 3. As to the effect of detergent on the biochemical composition of biomembrane, the content of carnitine, in E.coli, and B.cereus, the content of structural protein and phospholipid were decreased by treatment of sodium dodecyl sulfate and structural protein was denatured by palmitoyl choline. 4. The profile of membrane protein revealed that the bacterial membrane were composed of various proteins. By dint of this result, some of membrane proteins were solubilized or changed to small molecules by the treatment of sodium dodecyl sulfate and palmitoyl choline, in E.coli and membrane protein of the biomembrane by treatment of sodium dodecyl sulfate, sodium deoxy cholate, palmitoyl choline, and palmitoyl carnitine were confirmed to be different profile as compared with those of the control, in B. cereus. Therefore, it is suggested that sodium dfodecyl sulfate and palmitoyl choline soulbilized biomembranes or inhibited membrane transport and that palmitoyl carnitine and sodium deoxy cholate were used as an energy source or stimulating the membrane transport, in E.coli. And, it is suggested that all of detergents were inhibited biomembrane synthesis, expet saponin, in B.cereus.

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Ectopic Expression of Wild Rice OgGRP Gene Encoding a Glycine Rich Cell Wall Protein Confers Resistance to Botrytis cinerea Pathogen on Arabidopsis

  • Jeon, Eun-Hee;Chung, Eun-Sook;Lee, Hye-Young;Pak, Jung-Hun;Kim, Hye-Jeong;Lee, Jai-Heon;Moon, Byung-Ju;Jeung, Ji-Ung;Shin, Sang-Hyun;Chung, Young-Soo
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.193-198
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    • 2009
  • A full-length cDNA of OgGRP gene encoding a glycinerich cell wall protein was isolated from wild rice (Oryza grandiglumis). Deduced amino acid sequences of OgGRP are composed of 148 amino acids (16.3 kDa), and show 85.9% homology with Osgrp-2 (Oryza sativa). RT-PCR analysis showed that RNA expression of OgGRP was regulated by defense-related signaling chemicals, such as cantharidin, endothall, jasmonic acid, wounding, or yeast extract treatment. In relation to pathogen stress, the function of OgGRP was analyzed in OgGRP over-expressing Arabidopsis thaliana. Overexpression of OgGRP in Arabidopsis contributed to moderate resistance against fungal pathogen, Botrytis cinerea, by lowering disease rate and necrosis size. In the analysis of the transgenic Arabidopsis lines to check the change of gene expression profile, induction of PR1, PR5 and PDF1.2 was confirmed. The induction seemed to be caused by the interaction of ectopic expression of OgGRP with SA-and JA-dependent signaling pathways.

Immunochemical Localization of Tetrahydrocannabinol (THC) in Chemically Fixed Glandular Thrichomes of Cannabis (Cannabaceae)

  • Eun Soo Kim;Paul G. Mahlberg
    • Animal cells and systems
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    • v.3 no.2
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    • pp.215-219
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    • 1999
  • Monoclonal antibody for delta-9-tetrahydrocannabiol (THC Ab), conjugated with protein A-gold, was employed as a probe to detect THC localization in the gland and subjacent cells of chemically fixed bracts of Cannabis. THC was detected in the outer wall of the disc cells, fibrillar matrix, the surface feature of secretory vesicles, and sheath throughout development of the secretory cavity. The probe was absent from vesicles. Label was also present in anticlinal walls of disc cells and walls of dermal and mesophyll cells. Little or no THC Ab was present in disc cells and none were detected in control tissues. This distribution pattern of THC Ab was similar to that in tissues prepared by high pressure cryofixation-cryosubstitution. Consistent association of THC with wall and wall-derived materials suggests that cannnabinoids are synthesized outside the plasma membrane and bound to a wall component, where-upon they are transported to the cavity with wall materials released from the disc cell wall during development of the secretory cavity.

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Changes of Protein Bodies in Endosperm Cells during Embryo Development of Ginseng (Panax ginseng C.A. Meyer) Seeds - Seeds with Red Seed Coat and Indehiscent Seeds - (인삼(Panax ginseng C.A. Meyer) 종자의 배발달에 따른 배유세포의 단백과립 변화 - 홍숙 및 미개갑 종자 -)

  • 유성철
    • Journal of Plant Biology
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    • v.35 no.1
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    • pp.45-51
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    • 1992
  • The changes of protein bodies in endosperm cells of both seeds with red seed coat and indehiscent seeds of Panax ginseng C.A. Meyer have been investigated in relation to the embryo development. In the early stage of seeds with red seed coat, spherical spherosomes were distributed in endosperm cells. Protein bodies were formed from vacuoles containing the storage protein. Cell organelles were hardly observed in the cytoplasm. In the late stage of the seed with red seed coat, the endosperm was filled with spherosomes and protein bodies. The protein bodies consisted of amorphous inclusions with high electron density or proteinaceous matrix with even electron density. In the seed of in dehiscence, the protein body in endosperm cells contained globoids and protein crystalloids. The globoid of protein body had a electron dense materials. Umbiliform layer was formed between embryo and endosperm. The deformation patterns of endosperm cell wall and the cellulose microfibril were observed in endosperm cells near the umbiliform layer. Umbiliform layer consisted of lipid body and autolyzed cell debris. The protein body of endosperm cell near the umbiliform layer showed various degenerative patterns, and so electron density of proteinaceous matrix was gradually decreased.reased.

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Delivery of Protein into Microalgae by the Digital Electroporation (디지털 전기천공을 이용한 미세조류 내 단백질 전달 연구)

  • Im, Do Jin
    • Korean Chemical Engineering Research
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    • v.56 no.1
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    • pp.79-84
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    • 2018
  • In the present study, we performed electroporation to deliver protein into microalgae using previously developed digital electroporation system. Green fluorescence protein was successfully delivered into a live microalgae cell nucleus without cell wall removal. By investigating the effects of applied voltage on the protein delivery efficiency, optimal electroporation electric field condition was found (960 V/cm). We also investigated the delivery of Yo-Pro-1 into cell to examine the size effects of delivered materials and found that there is little size effects on the optimal condition. Finally, the implications of the present results and future work are discussed.