• Title/Summary/Keyword: hydrolytic enzyme

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(1-3, 1-4)-$\beta$-Glucan and Starch Contents and Their Hydrolytic Enzyme Activities in Developing Barley Kernels (등숙 중인 보리 종실중 (1-3, 1-4)-$\beta$-Glucan과 전분 함량 및 이들의 가수분해효소 활성)

  • 윤성중;박상래;유남희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.4
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    • pp.403-409
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    • 1997
  • To obtain information on the accumulation of (1-3, 1-4)-$\beta$-glucans during kernel maturation, (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities were determined in developing kernels of the two Korean cooking barley varieties, Neulssalbori and Saessalbori. (1-3, 1-4)-$\beta$-Glucan contents in kernels at 5 and 10 days after anthesis(DAA) were very low and the contents increased rapidly in kernels at 15 to 25 DAA. (1-3, 1-4)-$\beta$-Glucan content in kernels at harvest was about 3.5 to 4% of kernel dry matter. (1-3, 1-4)-$\beta$-Glucanase activities were relatively higher in younger kernels but the levels of the activity were very low compared with those in germinating kernels. A significant negative correlation was observed between (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities. Low levels of (1-3, 1-4)-$\beta$-glucanase activites in kernels at 15 to 30 DAA, however, may indicate that (1-3, 1-4)-$\beta$-glucanases have little effect on the final content of (1-3, 1-4)-$\beta$-glucans in barley kernels. Starch contents and $\alpha$-amylase activities were also determined in developing barley kernels. Starch contents increased rapidly as kernels matured and the content at harvest was about 60% of kernel dry matter. Relativley higher levels of $\alpha$-amylase activities in kernels at the earlier developmental stage decreased rapidly as kernels matured.

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Isolation and Identification of Low Molecular Weight Compounds Produced by Bacillus subtilis HJ927 and Their Biocontrol Effect on the Late Blight of Pepper (Capsicum annuum L.) (Bacillus subtilis HJ927에 의해 생성된 화합물의 분리, 동정 및 고추(Capsicum annum L.) 역병방제 효과)

  • Lee, Hyun-Jin;Park, Keun-Hyung;Shim, Jae-Han;Park, Ro-Dong;Kim, Yong-Woong;Hwang-Bo, Hoon;Cho, Jeung-Yong;Kim, Young-Cheol;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.25-31
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    • 2005
  • A soil bacterium, Bacillus subtilis HJ927, exhibiting strong antagonistic property against pathogenic fungi was isolated from pepper fields infested with Phytophthora capsici. Pepper plants inoculated with P. capsici revealed severe root mortality while plants co-inoculated with B. subtilis HJ927 and P. capsici showed drastically reduced root mortality. Low molecular weight substances released by B. subtilis HJ927 mediated the plant protective effect. The anti-fungal compounds released by B. subtilis HJ927 were identified as 3-methylbutyric acid, 2-methylbutyric acid, and methyl 2-hydroxy, 3-phenylpropanoate by high-performance liquid chromatography and gas chromatography-mass spectrometry. In addition to these compounds, B. subtilis HJ927 also produced ${\beta}$-1,3-glucanase, a hydrolytic enzyme implicated in antifungal activity.

Involvement of Growth-Promoting Rhizobacterium Paenibacillus polymyxa in Root Rot of Stored Korean Ginseng

  • Jeon, Yong-Ho;Chang, Sung-Pae;Hwang, In-Gyu;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.881-891
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    • 2003
  • Paenibacillus polymyxa is a plant growth-promoting rhizobacterium (PGPR) which can be used for biological control of plant diseases. Several bacterial strains were isolated from rotten roots of Korean ginseng (Panax ginseng C. A. Meyer) that were in storage. These strains were identified as P. polymyxa, based on a RAPD analysis using a P. polymyxa-specific primer, cultural and physiological characteristics, an analysis utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME), and the 16S rDNA sequence analysis. These strains were found to cause the rot in stored ginseng roots. Twenty-six P. polymyxa strains, including twenty GBR strains, were phylogenetically classified into two groups according to the ERIC and BOX-PCR analyses and 16S rDNA sequencing, and the resulting groupings systematized to the degrees of virulence of each strain in causing root rot. In particular, highly virulent GBR strains clustered together, and this group may be considered as subspecies or biovar. The virulence of the strains seemed to be related to their starch hydrolysis enzyme activity, but not their cellulase or hemicellulase activity, since strains with reduced or no starch-hydrolytic activity showed little or no virulence. Artificial inoculation of the highly virulent strain GBR-1 onto the root surfaces of Korean ginseng resulted in small brown lesions which were sunken and confined to the outer portion of the root. Ginseng root discs inoculated in vitro or two-year-old roots grown in soil drenched with the inoculum developed significant rot only when the inoculum density was $10^{6}-10^{7}$ or more colony-forming units (CFU) per ml. These results suggest that P. polymyxa might induce ginseng root rot if their population levels are high. Based on these results, it is recommended that the concentration of P. polymyxa should be monitored, when it is used as a biocontrol agent of ginseng, especially in the treatment of stored roots.

Effect of Exogenous Fibrolytic Enzyme Application on the Microbial Attachment and Digestion of Barley Straw In vitro

  • Wang, Y.;Ramirez-Bribiesca, J.E.;Yanke, L.J.;Tsang, A.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.66-74
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    • 2012
  • The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

Activities of Hydrolytic Enzymes in Barley Malts Prepared under Different Germination Conditions (발아조건별 당화용 엿기름의 분해효소 활성도)

  • Mok, Chul-Kyoon;Lee, Young-Tack
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.324-329
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    • 1999
  • Barley malts were prepared at 15, 18 and $21^{\circ}C$ for $3{\sim}6$ days, and assayed for ${\beta}-glucanase$, ${\alpha}-amylase$ and ${\beta}-amylase$ activities. ${\beta}-Glucanase$ activity increased markedly during earley germination and reached maximum at the 6th day of germination. ${\beta}-Glucanase$ activity in six-rowed barley malt was much higher than that in two-rowed malt. ${\beta}-Glucanase$ activity was associated with reduction in ${\beta}-glucan$ content during germination. ${\beta}-amylase$ activity was also considerably higher in two-rowed barley, and increased continuously during 6-day germination. ${\beta}-Amylase$ activity was the lowest at $15^{\circ}C$, the highest at $18^{\circ}C$, and intermediate at $21^{\circ}C$ of germination temperature. Considerable amount of ${\beta}-amylase$ was detected in ungerminated raw barley, and this enzymatic activity tended to increase during 6-day germination. Diastatic power, measure of starch-saccharifying enzyme, in six-rowed malt was $1.4{\sim}1.6$ fold higher than in two-rowed malt. Germination at $18^{\circ}C$ for $5{\sim}6$ days was suggested to be the optimum condition for manufacturing good quality malts, in terms of enhanced starch-degrading enzymatic activity.

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In vivo metabolism of carbofuran in resistant and susceptible brown planthoppers, Nilaparvata lugens $St{\aa}l$ (저항성 및 감수성 벼멸구 체내에서의 카보후란 대사)

  • Yoo, Jai-Ki;Ahn, Yong-Joon;Shono, Toshio;Lee, Si-Woo
    • The Korean Journal of Pesticide Science
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    • v.2 no.2
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    • pp.75-82
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    • 1998
  • This study was conducted to find out the biochemical or metabolic resistance mechanism of brown planthopper (BPH) to carbofuran. Differences between resistant ($LD_{50};\;20.3{\mu}g/g$) and susceptible strains($LD_{50};\;0.3{\mu}g/g$) were shown. The amounts of carbofuran metabolite, benzofuranol, and the origin, not developed by Thin Layer Chromatography, were much more in the susceptible strain. But the mother compound, carbofuran, was much more in the resistant strain. The tendencies of metabolism one and three hours after treatment were similar in both strains except for the amounts of metabolites described above. From the study, it is supposed that hydrolytic enzyme, esterase, changes its role from cleaving the esteric bond of carbofuran to making conjugates with carbofuran. This seems to be the main resistance mechanism of BPH to carbofuran. Oxidase and transferase may play little or no role in resistance mechanism. Oxidative and transferring enzymes gave no effects on the metabolism of carbofuran in the resistant strain compared with the susceptible strain.

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Journal of Ginseng ResearchHighly regioselective biotransformation of ginsenoside Rb2 into compound Y and compound K by β-glycosidase purified from Armillaria mellea mycelia

  • Kim, Min-Ji;Upadhyaya, Jitendra;Yoon, Min-Sun;Ryu, Nam Soo;Song, Young Eun;Park, Hee-Won;Kim, Young-Hoi;Kim, Myung-Kon
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.504-511
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    • 2018
  • Background: The biological activities of ginseng saponins (ginsenosides) are associated with type, number, and position of sugar moieties linked to aglycone skeletons. Deglycosylated minor ginsenosides are known to be more biologically active than major ginsenosides. Accordingly, the deglycosylation of major ginsenosides can provide the multibioactive effects of ginsenosides. The purpose of this study was to transform ginsenoside Rb2, one of the protopanaxadiol-type major ginsenosides, into minor ginsenosides using ${\beta}$-glycosidase (BG-1) purified from Armillaria mellea mycelium. Methods: Ginsenoside Rb2 was hydrolyzed by using BG-1; the hydrolytic properties of Rb2 by BG-1 were also characterized. In addition, the influence of reaction conditions such as reaction time, pH, and temperature, and transformation pathways of Rb2, Rd, F2, compound O (C-O), and C-Y by treatment with BG-1 were investigated. Results: BG-1 first hydrolyzes 3-O-outer ${\beta}$-$\text\tiny{D}$-glucoside of Rb2, then 3-O-${\beta}$-$\text\tiny{D}$-glucoside of C-O into C-Y. C-Y was gradually converted into C-K with a prolonged reaction time, but the pathway of Rb2 ${\rightarrow}$ Rd ${\rightarrow}$ F2 ${\rightarrow}$ C-K was not observed. The optimum reaction conditions for C-Y and C-K formation from Rb2 by BG-1 were pH 4.0-4.5, temperature $45-60^{\circ}C$, and reaction time 72-96 h. Conclusion: ${\beta}$-Glycosidase purified from A. mellea mycelium can be efficiently used to transform Rb2 into C-Y and C-K. To our best knowledge, this is the first result of transformation from Rb2 into C-Y and C-K by basidiomycete mushroom enzyme.

In vivo Metabolism of Flupyrazofos into Plutella xylostella (Lepidoptera: Yponomeutidae) and Spodoptera exigua (Lepidoptera: Noctuidae) (배추좀나방과 파밤나방의 채내에서 Flupyrazofos의 대사)

  • Lee, Sang-Guei;Lee, Hoi-Seon;Hwang, Chang-Yeon;Han, Man-Jong;Park, Hyung-Man
    • The Korean Journal of Pesticide Science
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    • v.6 no.3
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    • pp.224-229
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    • 2002
  • In vivo metabolism study was carried out to find out the biochemical or metabolic tolerance mechanism between Diamond backmoth (DBM), Plutella xylostella and Beet armywarm (BAW), Spodoptera exigua to flupyrazofos. They showed some differences between the DBM and BAW. About 20% of flupyrazofos applied to the 3rd instar larvae of DBM was metabolized within 1 h and about 50% of that was metabolized within 4 h. The metabolites of flupyrazofos-oxon in 3rd instar larvae of DBM were increased 10 times more at 4 h than 1 h after application. The amounts of flupyrazol were nearly same between at 1 h and 4 h. The amount of unknown and origin increased 2 and 3 times more at 1 h than 4 h after application, respectively. In the 4th instar BAW larva, about 50% of flupyrazofos was metabolized within 1 h and about 70% of that was metabolized within 4 h. As metabolites, the amounts of flupyrazofos-oxon increased 2 times more at 4 h than 1 h after application. The amounts of flupyrazol increased 4 times more at 4 h than 1 h after application. The amount of unknown and origin increased 2.5 and 2 times more at 4 h than 1 h after application, respectively. From the study, it is supposed that hydrolytic enzyme, esterase, cleave the alkyl bond of flupyrazofos and conjugates with flupyrazofos. This seems to be the main tolerance mechanism of BAW to flupyrazofos.

Isolation and Characteristics of Bacteria Showing Biocontrol and Biofertilizing Activities (생물방제 및 생물비료 활성을 가지는 세균의 분리 및 특성)

  • Jung, Ho-Il;Kim, Keun-Ki;Park, Hyean-Cheal;Lee, Sang-Mong;Kim, Yong-Gyun;Kim, Hong-Sung;Lee, Cnung-Yeol;Son, Hong-Joo
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1682-1688
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    • 2007
  • To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% $Ca_3(PO_4){_2}$ as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from $3{\sim}639{\mu}g/ml$ depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.

Hydrolysis of Arabinoxylo-oligosaccharides by α-ʟ-Arabinofuranosidases and β-ᴅ-Xylosidase from Bifidobacterium dentium

  • Lee, Min-Jae;Kang, Yewon;Son, Byung Sam;Kim, Min-Jeong;Park, Tae Hyeon;Park, Damee;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.32 no.2
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    • pp.187-194
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    • 2022
  • Two α-ʟ-arabinofuranosidases (BfdABF1 and BfdABF3) and a β-ᴅ-xylosidase (BfdXYL2) genes were cloned from Bifidobacterium dentium ATCC 27679, and functionally expressed in E. coli BL21(DE3). BfdABF1 showed the highest activity in 50 mM sodium acetate buffer at pH 5.0 and 25℃. This exo-enzyme could hydrolyze p-nitrophenyl arabinofuranoside, arabino-oligosaccharides (AOS), arabinoxylo-oligosaccharides (AXOS) such as 32-α-ʟ-arabinofuranosyl-xylobiose (A3X), and 23-α-ʟ-arabinofuranosyl-xylotriose (A2XX), whereas hardly hydrolyzed polymeric substrates such as debranched arabinan and arabinoxylans. BfdABF1 is a typical exo-ABF with the higher specific activity on the oligomeric substrates than the polymers. It prefers to α-(1,2)-ʟ-arabinofuranosidic linkages compared to α-(1,3)-linkages. Especially, BfdABF1 could slowly hydrolyze 23,33-di-α-ʟ-arabinofuranosyl-xylotriose (A2+3XX). Meanwhile, BfdABF3 showed the highest activity in sodium acetate at pH 6.0 and 50℃, and it has the exclusively high activities on AXOS such as A3X and A2XX. BfdABF3 mainly catalyzes the removal of ʟ-arabinose side chains from various AXOS. BfdXYL2 exhibited the highest activity in sodium citrate at pH 5.0 and 55℃, and it specifically hydrolyzed p-nitrophenyl xylopyranoside and xylo-oligosaccharides (XOS). Also, BfdXYL2 could slowly hydrolyze AOS and AXOS such as A3X. Based on the detailed hydrolytic modes of action of three exo-hydrolases (BfdABF1, BfdABF3, and BfdXYL2) from Bf. dentium, their probable roles in the hemiceullose-utilization system of Bf. dentium are proposed in the present study. These intracellular exo-hydrolases can synergistically produce ʟ-arabinose and ᴅ-xylose from various AOS, XOS, and AXOS.