• Title/Summary/Keyword: cell wall protein

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Characteristics of Structural Proteins of Synechococcus sp. Cyanophage (Synechoscoccus sp. cyanophage 구조단백질의 특성)

  • Kim, Seung-Won;Kim, Min;Leem, Mi-Hyea;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.242-246
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    • 1997
  • The protein profile of Synechococcus sp. cyanophage was investigated employing SDS-PAGE. The phage appears to be composed of two major proteins of 97 and 52 kDa and at least seven minor proteins of 70, 65, 60, 40, 35, 28, and 6 kDa. It seems that each subunit is combined to form a multimer although any disulfide bond does not exist in the phage structure. Lytic activity of the phage particle against cell wall was detected around the 52 kDa on renaturing SDS-PAGE using heat-killed Micrococcus luteus cells as substrate. The activity has the optimal pH between 9 and 10, and slightly inhibited by EDTA.

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Solubilization of Tofu-Residue Using Multienzyme Derived from Aspergillus niger CF-34 (Aspergillus niger CF-34 효소를 이용한 두부 또는 두유비지의 가용화)

  • Kim, Kang-Sung;Park, Eun-Ha;Choi, Yeon-Bae;Kim, Kyo-Chang;Lee, Sang-Hwa;Sohn, Heon-Soo
    • Korean Journal of Food Science and Technology
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    • v.26 no.5
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    • pp.484-489
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    • 1994
  • Solubilization of plant ceil wall(tofu-residue) using enzyme complex obtained by Aspergillus niger CF-34 was attempted. The hydrolysis reaction was done at pH 4.0, $50^{\circ}C$, which were optimum pH and temperature of the enzyme, respectively. At the enzyme dosage of 2.5% (in terms of solid content of tofu-residue) and reaction time of 3 hr, the solubilizing percent of protein and carbohydrate were 62% and 50% respectively. Homogenization prior to enzyme reaction did not have much effect on tofu-residue solubilization. To improve solubility of tofu-residue, additional treatment such as alkali with 0.1% NaOH solution was found to be useful. The results showed that tofu-residue, which mainly consists of cell wall component of cellulose and hemicellulose, was not accessible to enzyme reaction and some prior treatment is required to enhance enzyme hydrolysis.

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Utilization of Putrescine by Streptococcus pneumoniae During Growth in Choline-limited Medium

  • Ware D.;Watt J.;Swiatlo E.
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.398-405
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    • 2005
  • Polyamines such as putrescine are small, ubiquitous polycationic molecules that are required for optimal growth of eukaryotic and prokaryotic cells. These molecules have diverse effects on cell physiology and their intracellular content is regulated by de novo synthesis and uptake from the environment. The studies presented here examined the structure of a putative polyamine transporter (Pot) operon in Streptococcus pneumoniae (pneumococcus) and growth of pneumococci in medium containing putrescine substituted for choline. RT-PCR experiments demonstrated that the four genes encoding the Pot system are co-transcribed with murB, a gene involved in an intermediary step of peptidoglycan synthesis. Pneumococci grown in chemically-defined media (CDM) containing putrescine without choline enter logarithmic phase growth after 36-48 hs. However, culture density at stationary phase eventually reaches that of choline-containing medium. Cells grown in CDM-putrescine formed abnormally elongated chains in which the daughter cells failed to separate and the choline-binding protein PspA was no longer cell-associated. Experiments with CDM containing radiolabeled putrescine demonstrated that pneumococci concentrate this polyamine in cell walls. These data suggest that pneumococci can replicate without choline if putrescine is available and this polyamine may substitute for aminoalcohols in the cell wall teichoic acids.

Ginsenoside Rg2 Inhibits Lipopolysaccharide-Induced Adhesion Molecule Expression in Human Umbilical Vein Endothelial Cell

  • Cho, Young-Suk;Kim, Chan Hyung;Ha, Tae-Sun;Lee, Sang Jin;Ahn, Hee Yul
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.2
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    • pp.133-137
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    • 2013
  • Vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), P- and E-selectin play a pivotal role for initiation of atherosclerosis. Ginsenoside, a class of steroid glycosides, is abundant in Panax ginseng root, which has been used for prevention of illness in Korea. In this study, we investigated the mechanism(s) by which ginsenoside Rg2 may inhibit VCAM-1 and ICAM-1 expressions stimulated with lipopolysaccharide (LPS) in human umbilical vein endothelial cell (HUVEC). LPS increased VCAM-1 and ICAM-1 expression. Ginsenoside Rg2 prevented LPS-mediated increase of VCAM-1 and ICAM-1 expression. On the other hand, JSH, a nuclear factor kappa B (NF-${\kappa}B$) inhibitor, reduced both VCAM-1 and ICAM-1 expression stimulated with LPS. SB202190, inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), and wortmannin, phosphatidylinositol 3-kinase (PI3-kinase) inhibitor, reduced LPS-mediated VCAM-1 but not ICAM-1 expression. PD98059, inhibitor of mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) did not affect VCAM-1 and ICAM-1 expression stimulated with LPS. SP600125, inhibitor of c-Jun N-terminal kinase (JNK), reduced LPS-mediated ICAM-1 but not VCAM-1 expression. LPS reduced IkappaB${\alpha}$ ($I{\kappa}B{\alpha}$) expression, in a time-dependent manner within 1 hr. Ginsenoside Rg2 prevented the decrease of $I{\kappa}B{\alpha}$ expression stimulated with LPS. Moreover, ginsenoside Rg2 reduced LPS-mediated THP-1 monocyte adhesion to HUVEC, in a concentration-dependent manner. These data provide a novel mechanism where the ginsenoside Rg2 may provide direct vascular benefits with inhibition of leukocyte adhesion into vascular wall thereby providing protection against vascular inflammatory disease.

Production of Single Cell Protein on Rice Straw and Their Utilities (섬유소를 이용한 단세포단백질의 생산 및 그 이용)

  • Chung, Yung-Gun;Kwon, Oh-Jin
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.496-501
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    • 1995
  • Experiments were carried out to find out the optimal condition of SCP produced by Cellulomonas sp. KL-6 and to evaluate nutritional value for the protein of this organism Intracellular- and extracellular proteins produced by this strain were estimated to be nearly maximum, $266\;{\mu}g/m{\ell}\;and\;37\;{\mu}g/m{\ell}$, in the medium containing 0.001% of thiamin after 5 days cultivation. When used rice straw as carbon source for the cell growth of this organism after crusing them by cutting mill, and treating them with 1.0% of NaOH and 10.0% of $NH_4OH\;at\;80^{\circ}C$ for 30 minutes and neutralizing continuatively them with 85% of $H_3PO_4$, SCP production rates were very increased to $1.63\;g/{\ell}$ (NaOH) and $1.47\;g/{\ell}$ (NH4OH), respectively than $0.5\;g/{\ell}$ produced in untreated rice straw. We compared their amino acids patterns with that of FAO provisional patterns. Amino acids content of strain KL-6 was excellents. However. when intended these cell mass to use in practical animal feeding test it would be advisable that destruction or lysis of cell wall should be done.

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Optimization of Solid State Fermentation of Mustard (Brassica campestris) Straw for Production of Animal Feed by White Rot Fungi (Ganoderma lucidum)

  • Misra, A.K.;Mishra, A.S.;Tripathi, M.K.;Prasad, R.;Vaithiyanathan, S.;Jakhmola, R.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.208-213
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    • 2007
  • The objective of the experiment was to determine the optimum cultural [moisture levels (55, 60 and 70%), days of fermentation (7, 14 and 21), temperature (25 and $35^{\circ}C$) of incubation)] and nutritional parameters (urea addition (0 and 2%) and variable levels of single super phosphate (0.25 and 0.50% SSP)) for bio-processing of the mustard (Brassica campestris) straw (MS) under solid-state fermentation (SSF) system. The performance of SSF was assessed in terms of favorable changes in cell wall constituents, protein content and in vitro DM digestibility of the MS. Sorghum based inoculum (seed culture) of Ganoderma lucidum to treat the MS was prepared. The 50 g DM of MS taken in autoclavable polypropylene bags was mixed with a pre-calculated amount of water and the particular nutrient in the straw to attained the desired levels of water and nutrient concentration in the substrate. A significant progressive increase in biodegradation of DM (p<0.001), NDF (p<0.01) and ADF (p<0.05) was observed with increasing levels of moisture. Among the cell wall constituents the loss of ADF fraction was greatest compared to that of NDF. The loss of DM increased progressively as the fermentation proceeded and maximum DM losses occurred at 28 days after incubation. The protein content of the treated MS samples increased linearly up to the day $21^{th}$ of the incubation and thereafter declined at day $28^{th}$, whereas the improvement in in vitro DM digestibility were apparent only up to the day $14^{th}$ of the incubation under SSF and there after it declined. The acid detergent lignin (ADL) degradation was slower during the first 7 days of SSF and thereafter increased progressively and maximum ADL losses were observed at the day $28^{th}$ of the SSF. The biodegradation of DM and ADL was not affected by the variation in incubation temperature. Addition of urea was found to have inhibitory effect on fungal growth. The effect of both the levels (0.25 and 0.50) of SSP addition in the substrate, on DM, NDF, ADF, cellulose and ADL biodegradation was similar. Similarly, the protein content and the in vitro DM digestibility remain unaffected affected due to variable levels of the SSP inclusion in the substrate. From the results it may be concluded that the incubation of MS with 60 percent moisture for 21 days at $35^{\circ}C$ with 0.25 percent SSP was most suitable for MS treatment with Ganoderma lucidum. Maximum delignification, enrichment in the protein content and improvement in in vitro DM digestibility were achieved by adopting this protocol of bioprocessing of MS.

ON FARM DEMONSTRATION OF VARIOUS STORAGE METHODS FOR UREA TREATED WHEAT STRAW

  • Khan, A.G.;Ullah, W.;Azim, A.;Ali, A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.3
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    • pp.281-285
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    • 1996
  • On farm demonstration of urea treatment (5 kg urea dissolved in 60 litre water/100kg) of straw was performed at 6 different sites and treated straw was stored by three different methods i.e., plastic covered, mud plastered and existing farmers technique (mud plastered on the top and open from sides) to determine the best storage method in field. Untreated and treated samples were taken after 5 week storage period and subjected to crude protein, crude fibre and cell wall constituents analysis. In situ dry matter digestibility of straw was measured by nylon bag technique in buffalo bulls. Crude protein content increased by 100 to 153 percent in treated straw stored by different methods. Maximum increase in crude protein of treated straw was noticed in mud plastered method. Urea treatment of straw resulted in significant decrease in crude fibre contents in all the storage methods. Treatment of straw enhanced the in situ digestibility by 25-49 percent and maximum digestibility (53%) was found in mud plastered storage method. It was concluded that the mud plastered storage method for urea treated straw was found to be the best at farm level.

Enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus (축우 부루셀라병의 ELISA 진단법에 관한 연구)

  • Lim, Yoon-kyu;Lee, Doo-sick;Park, Jun-hong;Yang, Ki-chun;Kim, Seung-ho;Kim, Kong-sick;Hyun, Kwan-jong;Kim, Woo-tack;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.131-135
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    • 1993
  • Enzyme-linked Immuno sorbent Assay (ELISA) for the serological diagnosis of Brucella abortus was developed and compared with plate agglutination test. Cell wall antigen was extracted from Brucella abortus 1119-3 by sonication and with a sodium deoxychlate solution. Optimum protein concentration of coating antigen were $0.4{\mu}g/100{\mu}{\ell}$ protein on each microtiter plate well. Horse radish peroxidase (HRP) labeled protein-G was used as a tracer of reacted antibodies. ELISA confirmed the agreeable results of 40 cases out of 43 cases by plate aggulutination test. ELISA diagnosed positive cases(10 out of 12) and negative cases (1 out of 12) with dubious sera by plate agglutination test. From this results ELISA could be used for the early diagnostic tools of Brucellosis in cattle.

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N-terminal GNBP homology domain of Gram-negative binding protein 3 functions as a beta-1,3-glucan binding motif in Tenebrio molitor

  • Lee, Han-Na;Kwon, Hyun-Mi;Park, Ji-Won;Kurokawa, Kenji;Lee, Bok-Luel
    • BMB Reports
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    • v.42 no.8
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    • pp.506-510
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    • 2009
  • The Toll signalling pathway in invertebrates is responsible for defense against Gram-positive bacteria and fungi, leading to the expression of antimicrobial peptides via NF-$\kappa$B-like transcription factors. Gram-negative binding protein 3 (GNBP3) detects beta-1,3-glucan, a fungal cell wall component, and activates a three step serine protease cascade for activation of the Toll signalling pathway. Here, we showed that the recombinant N-terminal domain of Tenebrio molitor GNBP3 bound to beta-1,3-glucan, but did not activate down-stream serine protease cascade in vitro. Reversely, the N-terminal domain blocked GNBP3-mediated serine protease cascade activation in vitro and also inhibited beta-1,3-glucan-mediated antimicrobial peptide induction in Tenebrio molitor larvae. These results suggest that the N-terminal GNBP homology domain of GNBP3 functions as a beta-1,3-glucan binding domain and the C-terminal domain of GNBP3 may be required for the recruitment of immediate down-stream serine protease zymogen during Toll signalling pathway activation.

Cloning, Sequencing, and Expression of the Gene Encoding a Multidomain Endo-$\beta$-1,4-Xylanase from Paenibacillus curdlanolyticus B-6, and Characterization of the Recombinant Enzyme

  • Waeonukul, Rattiya;Pason, Patthra;Kyu, Khin Lay;Sakka, Kazuo;Kosug, Akihiko;Mori, Yutaka;Ratanakhanokchai, Khanok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.277-285
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    • 2009
  • The nucleotide sequence of the Paenibacillus curdlanolyticus B-6 xyn10A gene, encoding a xylanase Xyn10A, consists of 3,828 nucleotides encoding a protein of 1,276 amino acids with a predicted molecular mass of 142,726 Da. Sequence analysis indicated that Xyn10A is a multidomain enzyme comprising nine domains in the following order: three family 22 carbohydrate-binding modules (CBMs), a family 10 catalytic domain of glycosyl hydrolases (xylanase), a family 9 CBM, a glycine-rich region, and three surface layer homology (SLH) domains. Xyn10A was purified from a recombinant Escherichia coli by a single step of affinity purification on cellulose. It could effectively hydrolyze agricultural wastes and pure insoluble xylans, especially low substituted insoluble xylan. The hydrolysis products were a series of short-chain xylooligosaccharides, indicating that the purified enzyme was an endo-$\beta$-1,4-xylanase. Xyn10A bound to various insoluble polysaccharides including Avicel, $\alpha$-cellulose, insoluble birchwood and oat spelt xylans, chitin, and starches, and the cell wall fragments of P. curdlanolyticus B-6, indicating that both the CBM and the SLH domains are fully functioning in the Xyn10A. Removal of the CBMs from Xyn10A strongly reduced the ability of plant cell wall hydrolysis. These results suggested that the CBMs of Xyn10A play an important role in the hydrolysis of plant cell walls.