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The Biocontrol Activity of Chromobacterium sp. Strain C-61 against Rhizoctonia solani Depends on the Productive Ability of Chitinase

  • Park, Seur-Kee;Lee, Myung-Chul;Harman, Gary E.
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.275-282
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    • 2005
  • A chitinolytic bacterium, Chromobacterium sp. strain C-61, was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of eggplant. In this study, the biocontrol activity and enzymatic characteristics of strain C-61 were compared with its four Tn5 insertion mutants (C61-A, -B, -C, and -D) that had lower chitinolytic ability. The chitinase activity of a 2-day old culture was about $76\%,\;49\%\;and\;6\%$ level in C61-A, C61-B and in C61-C, respectively, compared with that of strain C-61. The $\beta-N-acetylhexosaminidase$(Nahase) activity was little detected in strain C-61 but increased largely in C-61A, C61-B and C61-C. Activities of chitinase and Nahase appeared to be negatively correlated in these strains. Another mutant, C-61D, produced no detectable extracellular chitinase and Nahase. The in vitro and in vivo biocontrol activities of strain C-61 and its mutants were closely related to their ability to produce chitinase but not Nahase. No significant differences in population densities between strain C-61 and its mutants were observed in soil around eggplant roots. The results of SDS-PAGE and isoelectrofocusing showed that a major chitinase of strain C-61 is 54-kDa with pI of approximately 8.5. This study provides evidence that the biocontrol activity of Chromobacterium sp. strain C-61 against Rhizoctonia solani depends on the ability to produce chitinase with molecular weight of 54-kDa and pI of 8.5.

Quality Characteristics of Squid Sikhae by Preparation Method and Fermentation Conditions (오징어 식해의 제조 방법에 따른 품질 특성)

  • Lee Ye-Kyung;Park Bum-Do;Kim Soon-Dong
    • Journal of the East Asian Society of Dietary Life
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    • v.15 no.4
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    • pp.405-412
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    • 2005
  • Quality characteristics of squid-sikhae prepared by four different methods(SHM: sikhae method, SHM-LA; sikhae method added with L plantarum, MM; mixed method of sikhae method and salting method, MM-LA; MM method added with L plantarum) were investigated during fermentation at $20^{\circ}C$. The pHs of all the 6-days fermented sikhae samples were in the range of 4.01-3.76, meaning that there were no significant difference in pH according to the preparation methods. Number of total microbes(TM) were decreased, while the ratio of lactic acid bacteria against TM in SHM-LA and MM-LA was higher than those of SHM and MM. There were no differences in acid protease activity, while $NH_2-N$ content of SHM and MM were higher than those of SHM-LA and MM-LA. Amylase activity was the lowest in MM-LA. Proteins separated by SDS-PAGE belonged to 7-200 kDa, the major proteins (153<94<41 kDa) of the sikhae in all plots were disappeared at 6 days fermentation. In sensory evaluation, sour taste of MM was the highest, while it was the lowest in SHM-LA. Sweet taste, bitter taste, salty taste and hot taste were not significantly different Off-flavor was decreased in lactic acid bacteria added products. Scores of the softness and overall acceptability were the highest in SHM-LA. These results indicated that SHM-LA was the best method for the preparation of squid sikhae because of the enhancement of lactic acid fermentation and overall acceptability.

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The Structural Characterization of Recombinant Bovine Somatotropin Expressed in Escherichia coli (재조합 소성장호르몬의 구조적 특성)

  • 김정호;김훈주박은숙김준
    • KSBB Journal
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    • v.9 no.2
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    • pp.165-173
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    • 1994
  • In this paper we have described the structural characterization of recombinant bovine somatotropin produced in Escherichia coli. Recombinant bovine somatotropin consists of 191 amino acid residues with a calculated molecular weight of 21,802 Da. For fragmentation of recombinant bovine somatotropin, we have used trypsin, Staphylococcus aureus V8 pretease, CNBr, and mild acid hydrolysis method. Digestion and cleavage with these proteases and chemicals yielded peptides of various size for amino acid sequence determination. The N-terminal sequence analysis was carried out up to thirty residues. Because the design of the recombinant bovine somatotropin gene for expression was such that the coding sequence begins with an initiation codon, AUG, before Ala, the first amino acid of bovine somatotropin, we could expect the initial amino acid as N-formyl Met. But the first amino acid of this protein, expressed in E. coli cells as inclusion bodies, was Ala. And the amino acid composition of RP-HPLC purified recombinant bovine somatotropin was determined and no essencial difference was observed. The amino acid sequence of the recombinant bovine somatotropin was identical to that predicted from its recombinant gene. There was no processing or replacement of amino acid residues in recombinant bovine somatotropin expressed in E. coli. The hydropathy plot of recombinant bovine somatotropin revealed a hydrophobic region at the NH2-terminus and hydrophilic region at the COOH-terminus. The E. coli expression system is thought to be valuable for the expression of recombinant bovine somatotropin because protein was processed to remove the N-terminal Met residue by methionyl-aminopeptidase autonomously.

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Antiendometrial Antibodies in Peritoneal Fluid from Patients with Endometriosis (자궁내막증 환자의 복강액내 항자궁내막항체에 관한 연구)

  • Kim, Jung-Gu;Kim, Dong-Ho;Choi, Doo-Suck;Kim, Dae-Won;Moon, Shin-Yong;Kang, Soong-Beom;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.17-24
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    • 1998
  • We have previously demonstrated that specific antigens involved in autoimmunity in endometriosis may be endometrial proteins with molecular weight (mw) of 71, 92, and 103 kilodalton (kDa). The purposes of this study were to determine the incidence of IgG antibodies against these endometrial antigens in peritoneal fluid of patients with endometriosis and to evaluate the antigenic differences between the endometria of patients with and without endometriosis. Forty peritoneal fluid (PF) from 24 patients with endometriosis and 16 patients without endometriosis (control patients) were tested against endometrial protein from patients (n=8) with endometriosis and from control patients (n=10) by western blot. Fifteen (62.5%) of 24 PF samples from patients with endometriosis had specific Immunoglobuiin (Ig) G antibodies against one of three endometrial proteins with mw of 71, 92 and 103 kDa but none of PF samples from control patients had these antibodies. The electrophoretic pattern of endometrial proteins from patients with endometriosis was similiar to that from control patients. Furthemore there was no significant difference in specific PF Immunoglobulin G binding to endometrial proteins regardless of origin of these proteins. Our data indicate that specific humoral immune response can be found in PF of patients with endometriosis and that specific antigens inducing this immune response are present in human endometrium and that there is no antigenic difference between the endometria of patients with and without endometriosis.

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Partial Purification and Characterization of Superoxide Dismutase from Tomato (Lycopersicon esculentum) Fruit

  • Kumar, Sunil;Dhillon, Santosh;Singh, Dharam;Singh, Randhir
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.283-288
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    • 2004
  • Superoxide dismutase (SOD) from tomato (Lycopersicon esculentum Mill.) fruit was purified by ammonium sulphate precipitation, Sephadex G-100 and DEAE-cellulose column chromatographies. A 22 fold purification and an overall yield of 44% were achieved. The purified enzyme was a homodimer with Mr 37.1 kDa and subunit Mr 18.2 kDa as judged by SDS-PAGE. SOD showed $K_{m}$ values of 25 ${\times}$ 10$^{-6}$ M and 1.7 ${\times}$ 10$^{-6}$ M for nitroblue tetrazolium (NBT) and riboflavin as substrates, respectively. The enzyme was thermostable upto 5$0^{\circ}C$ and exhibited pH optima of 7.8. The effect of metal ions and some other compounds on enzyme activity was studied. $Co^{2+}$ and $Mg^{2+}$ were found to enhance relative enzyme activities by 27 % and 73 %, respectively, while M $n^{2+}$ inhibited the SOD activity by 64%. However, $Ca^{2+}$ and C $u^{2+}$ had no effect on enzyme activity. Other compounds like $H_2O$$_2$ and Na $N_3$ inhibited enzymatic activities by 60% and 32%, respectively, while sodium dodecyl sulphate (SDS), chloroform plus ethanol and $\beta$-mercaptoethanol had no effect on the activity of SOD. of SOD.

Physico-chemical Properties and Changes of Sarcoplasmic Protein Bands of Chicken Meat Cuts with or without Salt during Cooking Temperatures (식염첨가 유무에 따른 계육의 부위별 가열온도에 따른 이화학적 성상과 근장 단백질 밴드의 변화)

  • Kim, Soo-Hee;Chin, Koo-Bok
    • Journal of Animal Science and Technology
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    • v.49 no.2
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    • pp.269-278
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    • 2007
  • This study was performed to measure the pH, proximate composition, physicochemical properties, changes of protein bands, Hunter color values and endpoint cooking temperature of chicken leg and breast muscles during cooking from 64 to 74℃ with 2℃ increments. Chicken leg had higher pH, moisture and fat contents (%) and lower protein solubility(P<0.05) than chicken breast. Although the cooking losses(CLs, %) of chicken muscles increased with increased cooking temperature, the addition of 2% salt did not affect CL. The redness values of chicken leg without 2% salt were higher than chicken breast, however, the addition of 2% salt reduced the differences of the redness. Protein solubility decreased with increased cooking temperatures and were not affected by the addition of salt, and no further changes were observed higher than 68℃. Protein bands having the molecular weights of 66 and 54kDa were disappeared in the chicken leg at the cooking temperatures of 66~70℃, whereas 66, 54 and 34kDa in the chicken breast. These protein bands could be used potential indicators to determine the endpoint cooking temperature in chicken muscles.

Effect of an Acid pH Shock on Physiological Changes of Chlamydomonas acidophila (Chlorophyta), UTCC 122 (pH 충격에 의한 Chlamydomonas acidophila (Chlorophyta), UTCC 122의 생리적 변화에 관한 연구)

  • Lee, Kyung;Ki, Jang-Seu;Kim, Say-Wa;Han, Myung-Soo;Choi, Young-Kil;Yoo, Kwang-Il
    • Korean Journal of Ecology and Environment
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    • v.35 no.3 s.99
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    • pp.145-151
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    • 2002
  • The effect of low pH on physiological changes was studied with the acidophilic green alga, Clamydomonas acidophila, UTCC 122. The growthrates (${\mu}$) were identical, $0.5{\sim}0.7\;day^{-1}$, at pH 3.7${\sim}$6.7 and no significantly different (ANOVA, p =0.134), showing cell volume reduced gradually as they were growing, whereas that at pH 2.7 was falling to zero and cell volume increased dramatically. Chlorophyll a concentration of the cultures incubated for one day was $191{\sim}255\;pg\;cell^{-1}$, after then it declined from $60{\sim}103\;pg\;cell^{-1}$ at pH 3.7${\sim}$6.7 except $210\;pg\;cell^{-1}$ at pH 2.7, which was directly related with cell volume. External carbonic anhydrase (CA) activity was varied from1.1 to$3.7{\times}10^{-4}\;E.U.\;mm^{-2}$, showing the gradualincrease during culture, except at 2.7 and pH 5.7. However there was not found any relationship among the pH gradient cultures. CA molecular mass of C. acidophila was 29 kBa, and concentration of that was identical in all cultures. The proteins of 41 kDa and 63 were not or very faintly expressed in low pH cultures, in contrast that of 17 kDa more expressed. In this work, we found that C. acidophila could live optimally within a wide range of acidic pH, and 17 kDa of unidentified protein might be concerned with tolerating in low acid environment.

Immuno stimulatory activities of Samul-tang, Sagunja-tang, Pamul-tang and Sipjeondaebo-tang in vitro (사물탕, 사군자탕, 팔물탕, 십전대보탕의 약리 활성 비교 연구)

  • Lee, Ho-Young;Ha, Hye-Kyung;Jung, Da-Young;Lee, Nam-Hun;Shin, Hyeun-Kyoo
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.4
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    • pp.41-51
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    • 2010
  • Objectives : Samul-tang(SM). Sagunja-tang(SG). Pamul-tang(PM) and Sipjeondaebo-tang(SJ) was used many diseases such as sterility. menstrual disorder. general prostration. recruitment in Korea. We investigated the immune stimulatory activities of SG. SM. PM and SJ in in vitro. Methods: For comparision for effective of SM. SG. PM and SJ. this study examined anti-inflammation(NO. PGE2 assay). anti-oxidation(DPPH assay) and immune response in in vitro assay. For immune response activities. this study used NO synthesis on RAW 264.7 cells, splenocyte proliferation and cytokine assay(IL-2, IL-4) in splenocyte. Results: The results showed that SG. SM. PM and SJ were no significant effect anti-oxidation and anti-inflammatory effects. For immune response. they showed the splenocyte proliferation and macrophage proliferation. We confirmed that they synthesised NO a dose-dependent manner significantly and secreted the IL-4. Conclusions : These results suggested the SG, SM, PM and SJ had immune stimulatory activity. A efficacy of immune response in them had produced similar results.

The Effect of Sonicated Extracts of Treponema Denticola and Treponema Lecithinolyticum on the Cytokine Secretion and Matrix Metalloproteinase Activation of Gingival Fibroblast (Treponema denticola와 Treponema lecithinolyticum의 분쇄액이 치은섬유아세포의 Cytokine 분비 및 Matrix metalloproteinase 활성에 미치는 영향)

  • Suh, Hye-Yuhn;Choi, Bong-Kyu;Choi, Seong-Ho;Cho, Kyoo-Sung;Kim, Chong-Kwan;Chai, Jung-Kiu
    • Journal of Periodontal and Implant Science
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    • v.29 no.4
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    • pp.979-995
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    • 1999
  • This study was investigated to observe the effect of Treponema denticola cell sonicates(TDC) and Treponema lecithinolyticum cell sonicates(TLC) on cytokine secretion and matix metalloproteinase-2(MMP-2) activation of cultured human gingival fibroblast. Several experiments were performed including $IL-1{\beta}$, IL-6 ELISA for the effect on the $IL-1{\beta}$, IL-6 secretion of human gingival fibroblast. Also gelatinase zymography and gelatin dissolubility test for the activation of MMP-2 secreted by gingival fibroblast. The results were as follows. 1. The effect of TDC and TLC on IL-6 secretion of human gingival fibroblast showed statistically significant increase of IL-6 secretion in the TDC and TLC treated group compared to no treatment group(p<0.05) . 2. The amount of $IL-1{\beta}$ secretion was below the lower limit and there was no difference in the $IL-1{\beta}$ secretion of gingival fibroblast between TDC, TLC treated group and no treatment group. 3. The active form of pro MMP-2 with 72 kDa molecular weight was activated in both TDC and TLC treated group and clear band was appeared at 62kDa site on the zymography. 4. Gelatin dissolubility of MMP-2 secreted by gingival fibroblast was higher in TDC and TLC treated group compared to no treatment group(p<0.05). 5. In the TDC treated group, serine protease of T. denticola affect gelatin dissolubility. But in the TLC treated group gelatin was degraded by only MMP secreted by gingival fibroblast. Regarding to the above results, TDC and TLC have an effect on the IL-6 secretion increase of human gingival fibroblast and appears to activate pro MMP-2 which degrades collagen.

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Environmental stress-related gene expression and blood physiological responses in olive flounder (Paralichthys olivaceus) exposed to osmotic and thermal stress

  • Choi, Cheol-Young
    • Animal cells and systems
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    • v.14 no.1
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    • pp.17-23
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    • 2010
  • We isolated warm temperature acclimation-related protein 65-kDa (Wap65) cDNA from the liver of olive flounder and investigated the mRNA expression of Wap65 and HSP70 in olive flounder exposed to osmotic (17.5, 8.75, and 4 psu) and thermal stress (25 and $30^{\circ}C$). The mRNA expression of Wap65 and HSP70 was increased by thermal stress. The mRNA expression of HSP70 was also increased by osmotic stress, whereas no significant change in Wap65 expression was detected. These results indicate that Wap65 mRNA expression occurs specifically in response to increases in water temperature, but not in response to osmotic stress. Plasma cortisol levels were also increased by osmotic and thermal stress. We also utilized the stress hormone cortisol to examine whether Wap65 expression is thermal-stress-specific. Cortisol treatment increased HSP70 mRNA expression in vitro, but had no significant effect on Wap65 mRNA expression. Thus, thermal stress, but not osmotic stress, induces Wap65 expression.