• Title/Summary/Keyword: Plant Crude Enzymes

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The Production and Enzymatic Properties of Extracellular Chitinase from Pseudomonas stutzeri YPL-1, as a Biocontrol Agent

  • Lim, Ho-Seong;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.134-140
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    • 1994
  • An antagonistic bacterium Pseudomonas stutzeri YPL-1 liberated extracellular chitinase and $\beta$-1,3-glucanase which are key enzymes in the decomposition of fungal hyphal walls. The lytic enzymes caused abnormal swelling and retreating at the hyphal tips of plant pathogenic fungus Fusarium solani in a dual culture. Scanning electron microscopy revealed the hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. The production of chitinase and properties of a crude preparation of the enzyme from P. stutzeri YPL-1 were investigated. Peak of the chitinase activity was detected after 4 hr of cultivation. The enzyme had optimum temperature and pH of 50$^{\circ}C$ and pH 5.3, respectively. The enzyme was stable in the pH range of 3.5 to 6.0 up to 50$^{\circ}C$. The enzyme was significantly inhibited by metal compounds such as $HgCl_2$, but was stimulated by $CoCl_2$. P. stutzeri YPL-1 produced high levels of the enzyme after 84 hr of incubation. Among the tested carbon sources, chitin was the most effective for the enzyme production, at the concentration level of 3%. As a source of nitrogen, peptone was the best for the enzyme production, at the concentration level of 4%. The maximum amount of enzyme was produced by cultivating the bacterium at a medium of initial pH 6.8.

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The Relative Identification of C. officinale and L. chuanxiong by PCR-Mediated Fingerprinting (천궁류(川芎類) 한약재의 유전자 감식 연구)

  • Choi, Ho-Young;Kim, Dong-Wook;Kim, Dong-Eun;Suh, Young-Bae;Ham, In-Hye
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.151-161
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    • 2005
  • Objectives : Our research purpose is to establish the standard identification analysis on C. officinale and L. chuanxiong in Korea and China by PCR-mediated fingerprinting. Methods : The Restriction Fragment Length Polymorphism (RFLP) and Randomly Amplified Polymorphic DNA (RAPD) method was used on Internal Transcribed Spacer (ITS) regions and rbcL regions to compare and discriminate genes extracted from crude drugs as C. officinale and L. chuanxiong in Korea and China. Results : L. chuanxiong Korea and China have very similar polymorphism, whereas L. chuanxiong in Korea and C. officinale have very different polymorphism in RFLP. And restriction enzymes AluI and SacI forms the specific fragment band only in C. officinale, they can be used as RFLP marker on ITS regions to discriminate among the species. Conclusions : The results could be applied in discriminating crude drugs among C. officinale and L. chuanxiong in Korea and China. Also they could be used in controlling drug quality, preserving medicinal plants, and improving plant description.

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Effect of Dioscorea Rhizoma on Gastritis by Acute Gastric Ulcer in Rats (산약이 흰쥐의 급성 위궤양에 의한 위장 장애에 미치는 효과)

  • Park, Young Mi;Lim, Jae Hwan;Seo, Eul Won
    • Korean Journal of Plant Resources
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    • v.28 no.1
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    • pp.1-8
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    • 2015
  • In order to verify the effects of Dioscorea Rhizoma on gastrointestinal damages, we investigated the protective role of the crude extracts on induced gastric ulceration in rats. Rats administered of Dioscorea Rhizoma extracts showed gradually decreased congestion and hemorrhage, but control group did not show any symptom in gastric tissue. Moreover, Dioscorea Rhizoma extract had a role in lowering gastrin and histamine levels in gastric ulcer rats, thereby inhibiting the gastric tissue damages. Excessive production of malondialdehyde shown in gastric ulcer rats was declined in all rats administered with Dioscorea Rhizoma extract as well as the levels of SOD and GPX surged by acute gastric ulcer. Also, the increased activity of CAT showed an effect in activation of antioxidant enzyme to normal state. All data suggest that Dioscorea Rhizoma extract was verified to be highly effective resource in improving the gastrointestinal function of rats by preventing from gastric tissue damage in acute gastric ulcer and restoring the activities of plasma substances and antioxidant enzymes.

Partial Purification of Lectin from Mycoparasitic Species of Trichoderma

  • Singh, Tanuja;Saikia, Ratul;Arora, Dilip K.
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.301-309
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    • 2005
  • Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

Protective Effects of Crude Mucin and Saponin from Dioscorea Rhizoma on Gastric Ulcer Induced by Alcohol in Rats (산약의 조추출 mucin과 saponin이 급성 위궤양이 유도된 흰쥐에 미치는 보호 효과)

  • Park, Young Mi;Lim, Jae Hwan;Jeong, Hyung Jin;Seo, Eul Won
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1200-1208
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    • 2014
  • The aim of this study was to investigate the effect of crude mucin and saponin from Dioscorea Rhizoma on acute gastric ulcers in rats. The gastric ulcer group (GU group) and mucin-applied group (DR-M group) exhibited serious bleeding of the mucous membrane of the stomach due to the ulcers, as well as blood congestion for three days. The saponin-applied group (DR-S group) exhibited less mucous membrane bleeding, and reddened and inflamed membranes recovered dramatically within 24 hours. After developing an acute pgastric ulcer, the tissues of the stomach, intestine, and liver in the control group and the DR-M group exhibited edema in the submucous membrane, as well as serious bleeding. However, the DR-S group recovered quickly from mucous membrane bleeding due to gastric ulcer. The DR-M group did not show any notable changes in serum formation or activity of antioxidant enzymes compared to the GU rats. Increased AST and ALT activities were detected from the first day with saponin application in the gastric ulcer rats. As the AST and ALT activities decreased, the gastric ulcers recovered with the increased activities of the antioxidant enzymes. Accordingly, this study suggest that mucin in Dioscorea Rhizomahas no effect on the recovery of damaged stomachs due to gastric ulcers, but saponin is mainly responsible for decreasing tissue damage by activating antioxidant enzymes.

Isolation of cellulosic biomass degrading microorganisms from different sources for low cost biofuel production

  • Sheikh, M. Mominul Islam;Kim, Chul-Hwan;Lee, Ji-Yong;Yeasmin, Shabina;Park, Hyeon-Jin;Kim, Gyeong-Chul;Kim, Sung-Ho;Kim, Jae-Won
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2011.04a
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    • pp.81-91
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    • 2011
  • Current fuel ethanol research and development deals with process engineering trends for improving biotechnological production of ethanol. Recently, a large amount of studies regarding the utilization of lignocellulosic biomass as a good feedstock for producing fuel ethanol is being carried out worldwide. The plant biomass is mainly composed of cellulose, hemicellulose and lignin. The main challenge in the conversion of biomass into ethanol is the complex, rigid and harsh structures which require efficient process and cost effective to break down. The isolation of microorganisms is one of the means for obtaining enzymes with properties suitable for industrial applications. For these reasons, crude cultures containing cellulosic biomass degrading microorganisms were isolated from rice field soil, cow farm soil and rotten rice straw from cow farm. Carboxymethyl cellulose (CMC), xylan and Avicel (microcrystalline cellulose) degradation zone of clearance on agar platefrom rice field soil resulted approximately at 25 mm, 24 mm and 22 mm respectively. As for cow farm soil, CMC, xylan and Avicel degradation clearancezone on agar plate resulted around at 24mm, 23mm and 21 mm respectively. Rotten rice straw from cow farm also resulted for CMC, xylan and Avicel degradation zone almost at 24 mm, 23 mm and 22 mm respectively. The objective of this study is to isolatebiomass degrading microbial strains having good efficiency in cellulose hydrolysis and observed the effects of different substrates (CMC, xylan and Avicel) on the production of cellulase enzymes (endo-glucanase, exo-glucanase, cellobiase, xylanase and avicelase) for producing low cost biofuel from cellulosic materials.

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Increased Antifungal Activity with Genetic Development of Antagonistic Pseudomonas stutzeri YPL-1 against Fusariym solani (식물근부균 Fusarium Solani에 길항하는 생물방제균 Pseudomonas stutzeri YPL-1의 유전공학적 개발)

  • 임호성;김상달
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.437-441
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    • 1990
  • For the genetic development of more powerful antagonistic Pseudomom - YPL-1 as a biocontxol agent against soilborne plant pathogenic Fuaarium solani causing root rot of many important crops, mutants improving the productivity of chitinase were obtained by mutation with UV radiation or NTG treatment, P. stutzeri YPL-M26 (UV mutant) and P. stutzeri YPL-MI78 (NTG mutant) could improve the productivity of chitinase by 2.5 and 2.0 times, and its antifungal activity by 1.7 and 1.5 times, respectively. The antifungal mechanism of P. stutzeri YPL-M26 was caused by lysis of the fungal cell wall by hydrolytic enzymes such as chitinase. The antifungal activity of crude chitinase of P. stutzeri YPLM26 on the mycelial growth of F. solani was observed to be much higher than that of the original strain. The enzymes produced by P. stutzeri YPL-M26 were the same as the original strain in enzymatic properties such as optimal pH and temperature.

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A Specific Pullulanase for ${\alpha}$-1,6-Glucosidic Linkage of Glucan from Thermus caldophilus

  • Moon-Jo Lee;June-Ki Kim;Kyung-Soo Nam;Jin-Woo Park;Cher-Won Hwang;Dong-Soo Kim;Cheorl-Ho Kim
    • Journal of Life Science
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    • v.9 no.1
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    • pp.26-34
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    • 1999
  • A thermostable pullulanase has been isolated and purified from Thermus caldophilus GK-24 to a homogeneity by gel-filtration and ion-exchange chromatography. The specific activity of the purified enzyme was 431-fold increase from the crude culture broth with a recovery of 11.4%. The purified enzyme showed $M_{r}$ of 65 kDa on denaturated and natural conditions. The pI of the enzyme was 6.1 and Schiff staining was negative, suggesting that the enzyme is not a glycoprotein. The enzyme was most active at pH 5.5. The activity was maximal at $75^{\cire}C$ and stable up to $95^{\cire}C$ for 30 min at pH 5.5. The enzyme was stable to incubation from pH 3.5 to pH 8.0 at $4^{\cire}C$ for 24hr. The presence of pullulan protected the enzyme from heat inactivation, the extent depending upon the substrate concentration. The activity of the enzyme was simulated by $Mn^{2+}$ ion, }$Ni^{2+}$, $Ca^{2+}$, $Co^{2+}$ ions. The enzyme hydrolyzed the ${\alpha}$-1,6-linkages of amylopectin, glycogens, ${\alpha}$, ${\beta}$-limited dextrin, and pullulan. The enzyme caused the complete hydrolysis of pullulan to maltotriose and the activity was inhibited by $\alpha$, $\beta$, or $\gamma$-cyclodextrins. The $NH_{2}$-terminal amino acid sequence [(Ala-Pro-Gln-(Asp of Tyr)-Asn-Leu-Leu-Xaa-ILe-Gly-Ala(Ser)] was compared with known sequences of various sources and that was compared with known sequences of various sources and that was different from those of bacterial and plant enzymes, suggesting that the enzymes are structurally different.

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Molecular Identification of Four Different α-amylase Inhibitors from Baru (Dipteryx alata) Seeds with Activity Toward Insect Enzymes

  • Bonavides, Krishna B.;Pelegrini, Patricia B.;Laumann, Raul A.;Grossi-De-Sa, Maria F.;Bloch, Carlos Jr.;Melo, Jorge A.T.;Quirino, Betania F.;Noronha, Eliane F.;Franco, Octavio L.
    • BMB Reports
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    • v.40 no.4
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    • pp.494-500
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    • 2007
  • The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of $\alpha$-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, $\alpha$-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis $\alpha$-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus $\alpha$-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed $\alpha$-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several $\alpha$-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species.

Effect of Tectorigenin Obtained from Pueraria thunbergiana Flowers on Phase I and -II Enzyme Activities in the Streptozotocin-induced Diabetic Rat

  • Choi, Jong-Won;Shin, Myung-Hee;Park, Kun-Young;Lee, Kyung-Tae;Jung, Hyun-Ju;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.9 no.4
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    • pp.235-240
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    • 2003
  • Tectorigenin has an apoptosis-inducing ability and immunosuppressive activity. We investigated the effect of tectorigenin on Phase I and II enzyme activities to elucidate the pharmacological action of the immunosuppressive tectorigenin in the diabetic rat. This compound was obtained from the hydrolysis of tetoridin isolated from the flower of Pueraria thumbergiana (Leguminosae). This crude drug (Puerariae Flos) has been used as a therapeutic for diabetes mellitus in traditional Korean medicine. Tecotrigenin inhibited the formation of malondialdehyde (MDA) and hydroxy radicals in serum and liver but promoted superoxide dismutase (SOD) activity. Low MDA contents and low xanthine oxidase and aldehyde oxidase activities were observed in the tectorigenin-treated rats, suggesting that such Phase I enzyme activities are the major source of lipid peroxidation. However, tectorigenin increased Phase II enzyme activities such as SOD, glutathione peroxidase and catalase, suggesting the activation of free radical-scavenging enzymes. The activities of tectorigenin were comparable to those of glibenclamide, which was employed as a positive control. These results suggest that tectorigenin may share some biological properties with glibenclamide in insulin-dependent-diabetes mellitus (IDDM).