• Title/Summary/Keyword: D-cellobiose

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$^{1}H-NMR$ Spectroscopic Evidence on the Glycosidic Linkages of the Transglycosylated Products of Low-Molecular-weight 1,4-$\beta$-D-Glucan Glucanohydrolase from Trichoderma koningii (Trichoderma koningii에서 분리한 저분자 1, 4-$\beta$-D-Glucan Glucanohydrolase의 반응산물에 대한 핵자기공명 스펙트럼 분석)

  • 맹필재;강사욱;정춘수;홍순우;하영칠;이영하;김재헌
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.304-308
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    • 1987
  • The mode of transglycosylation reaction observed during the action of low-molecular-weight 1,4-$\beta$-D-glucan glucanohydrolase (EC 3.2.1.4) purified from Trichoderma koningii ATCC 26113 was investigated using $^{1}H-NMR $spectroscopy. The H-1 proton resonances were analysed. After reaction of the enzyme with cellotriose, the reaction products were separated by high performance liquid chromatography. H-1 resonances of the products were consisted with those of cellobiose, cellotriose and cellotitraose, respectively. Therefore it was proved that all the reaction products formed by the action of the enzyme on cellooligosaccharides, including transglycosylation products, possess only H-NMR -1,4-glycosidic linkage(s).

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$^{1}$H-NMR spectroscopic evidence on the glycosidic linkages of the transglycosylated products of low-molecular-weight $\beta$-D-glucosidase from trichoderma koningii (Trichoderma koningii에서 분비되는 .$\beta$-D-glucosidase의 반응산물에 대한 핵자기공명분석)

  • 이헌주;정춘수;강사욱;하영칠
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.35-42
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    • 1989
  • The mode of transglycosylation reaction observed during the action of low-molecular-weigh $\beta$-D-glucosidase ($\beta$-D-glucoside glucohydrolase, EC3.2.1.21) purified from Trichoderma koningii ATCC 26113 was investigated using $^{1}H$-NMR spectroscopy. The enzyme was purified by the series of procedures including ammonium sulfate precipitation, and fractionations by column chromatographies on Bio-Gel P-150, DEAE-Sephadex A-50, and SP-Sephadex C-50. The final purification was performed by the band eluation after preparative polyacrylamide gel electrophoresis. The enzyme showed its molecular size of 78,000 through the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its isoelectric point of 5.80 through the analysis of analytical isoelectric focusing. The H-1 proton resonances were analyzed. After the reaction of the enzyme with cellobiose, the reaction products were separated by high performance liquid chromatography using refractive index detector. H-1 resonances of the products were consisted with those of gentiobiose [$\beta$-D-glucopyranosyl--(1,6)-D-glucopyranose], and cellotriose [$\beta$-D glucopyranosyl-(1,4)-$\beta$-D-glucopyranosyl]-(1,4)-D-glucopyranose] with minor resonances of sophorose [$\beta$-D-glucopyranosyl-(1,2)-D-glucopyranose], respectively.

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Isolation, Identification, and Characterization of Ornithine-Producing Enterococcus faecalis OA18 from Kefir Grain (케피어그레인으로 제조한 요쿠르트로부터 Enterococcus faecalis OA18 균주의 분리 및 특성규명)

  • Yu, Jin-Ju;Kim, Su-Gon;Seo, Kyoung-Won;Oh, Suk-Heung
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.218-224
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    • 2011
  • Lactic acid bacteria (LAB) OA18 was isolated from yogurt prepared by using Kefir Grain as a starter. The OA18 strain was a Gram-positive, cocci-type bacterium, and able to grow anaerobically with $CO_2$ production. The OA18 strain grew well on MRS broth supplemented with 50 mM arginine at $30-37^{\circ}C$ and pH of 7.0-9.0. The optimum temperature and pH for growth are $37^{\circ}C$ and pH 7.0. The isolate fermented ribose, D-glucose, cellobiose, D-trehalose, but not L-xylose, D-melibiose, and inositol. The 16S rRNA gene sequence of the isolate showed 99.8% homology with the Enterococcus faecalis 16S rRNA gene (Access no. AB012212). Based on the biochemical characteristics and 16S rRNA gene sequence analysis data, it was identified and named as E. faecalis OA18. The E. faecalis OA18 strain showed a high ornithine-producing capacity in the presence of arginine and also showed an antimicrobial activity against Streptomyces strains such as Streptomyces coelicolor subsp. Flavus, S. coeruleorubidus, S. coeruleoaurantiacus, S. coelicolor, S. coeruleoprunus. The cell growth of E. faecalis OA18 strain was maintained in MRS broth with a NaCl concentration of 0-7%.

Isolation and characterization of unrecorded yeast species from Korea in the families Debaryomycetaceae and Piskurozymaceae

  • Lee, Sang Eun;Oh, Hye Jin;Kim, Myung Kyum
    • Journal of Species Research
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    • v.10 no.4
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    • pp.344-349
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    • 2021
  • The purpose of this study was to isolate and identify wild yeasts from soil of Gyeongju city, and Haemadipsa rjukjuana of Gageodo Island, characterizing unrecorded yeast strains from Korea. The molecular analysis of the D1/D2 domain of 26S rDNA of yeast was performed using the Basic Local Alignment Search Tool (BLAST). No official report exists describing these three species: one species in the genus Candida, one species in the genus Debaryomyces, and one species in the genus Solicoccozyma. Candida saitoana YL9, Debaryomyces fabryi YL1, and Solicoccozyma terrea 20g9-1 are recorded for the first time from Korea. All three strains were oval shaped and polar binding, while positive for glucose, ᴅ-xylose, and ᴅ-cellobiose. Morphological, physiological, and biochemical properties are described in the species descriptions.

Hanseniaspora thailandica BC9 β-Glucosidase for the Production of β-ᴅ-Hexyl Glucoside

  • Phongprathet, Sujittra;Vichitphan, Kanit;Han, Jaehong;Vichitphan, Sukanda;Sawaengkaew, Jutaporn
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.579-587
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    • 2018
  • For biotechnological production of high-valued ${\beta}-{\text\tiny{D}}$-hexyl glucoside, the catalytic properties of Hanseniaspora thailandica BC9 ${\beta}$-glucosidase purified from the periplasmic fraction were studied, and the transglycosylation activity for the production of ${\beta}-{\text\tiny{D}}$-hexyl glucoside was optimized. The constitutive BC9 ${\beta}$-glucosidase exhibited maximum specific activity at pH 6.0 and $40^{\circ}C$, and the activity of BC9 ${\beta}$-glucosidase was not significantly inhibited by various metal ions. BC9 ${\beta}$-glucosidase did not show a significant activity of cellobiose hydrolysis, but the activity was rather enhanced in the presence of sucrose and medium-chain alcohols. BC9 ${\beta}$-glucosidase exhibited enhanced production of ${\beta}-{\text\tiny{D}}$-hexyl glucoside in the presence of DMSO, and 62% of ${\beta}-{\text\tiny{D}}$-hexyl glucoside conversion was recorded in 4 h in the presence of 5% 1-hexanol and 15% DMSO.

Enhancement of Biocontrol Activity of Antagonistic Chryseobacterium Strain KJ1R5 by Adding Carbon Sources against Phytophthora capsici

  • Kim, Yu-Seok;Jang, Bo-Ra;Chung, Ill-Min;Sang, Mee-Kyung;Ku, Han-Mo;Kim, Ki-Deok;Chun, Se-Chul
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.164-170
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    • 2008
  • Carbon utilization by Chryseobacterium strain KJ1R5 was studied to enhance its biocontrol activity against Phytophthora capsid. Chryseobacterium strain KJ1R5 has previously been shown to control Phytophthora blight of pepper (Capsicum annuum L.). Strain KJ1R5 could utilize carbon sources such as L-arabinose, D-cellobiose, ${\beta}-lactose$ and D-galactose well. P. capsici could utilize D-glucose well, showing the absorbencies ranged from 0.577 to 0.767 at 600nm. When 2% L-arabinose, which could only be utilized by the bio-control strain KJ1R5, was amended into the bacterial suspension, the efficacy of biological control increased. Among the amendments of various carbon sources into bacterial suspension, L-arabinose and D-(+)-glucose significantly enhanced biological control activity, resulting in a reduction of disease incidence to 6.9%, compared to 21.9% for the strain KJ1R5 alone and 81.3% for P. capsici inoculation alone, indicating that amendment with specific carbon sources could increase the biological control activity.

Purification and Characterization of Alkaline Invertase from the Hypocotyls of Mung Bean (Phaseolus raiatus L.) (녹두의 하배축에서 분양한 Alkaline lnvertase의 정제와 특성)

  • Young-Sang Kim
    • Journal of Plant Biology
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    • v.38 no.4
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    • pp.349-357
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    • 1995
  • The alkaline invertase ($\beta$-D-fructofuranoside fructohydrolase, EC 3.2.1.26) was isolated and characterized from the hypocotyls of mung bean (Phaseolus radiatus L.). The enzyme was purified by consecutive step using diethylaminoethyl (DEAE)-cellulose anion exchange, 1st Sephadex G-200, DEAE-Sephadex A50 and 2nd Sephadex G-200 chromatography. The overall purification was about 77-fold with a yield of about 6%. The finally purified enzyme exhibited a specific activity of about 48 $\mu$mol of glucose produced mg-1 protein min-1 at pH 7.0 and appeared to be a single protein by nondenaturing polyacrylamide gel electrophoresis (PAGE). The enzyme had the native molecular weight of 450 kD and subunits molecular weight of 63 kD and 38 kD as estimated by Sephadex G-200 chromatography and SDS-PAGE, respectively, suggesting that the enzyme is a heteromultimeric protein composed of two types of subunits. On the other hand, the enzyme appeared to be not a glycoprotein according to the results of Con A chromatography and glycoprotein staining. The enzyme had a Km for sucrose of 19.7 mM at pH 7.0 and maximum activity around pH 7.5. The enzyme was most active with sucrose as substrate, compared to raffinose, cellobiose, maltose and lactose. These results indicate the alkaline invertase is a $\beta$-fructofuranosidase.

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Studies on Isolation and Characterization of Anaerobic Bacteria from Gut of Holstein Cows and Korean Male Spotted Deer (꽃사슴과 Holstein 젖소의 장내 혐기성 박테리아의 분리 및 특성)

  • 박소현;이기영;안종호;장문백;김창현
    • Journal of Animal Science and Technology
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    • v.48 no.1
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    • pp.77-90
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    • 2006
  • The purpose of this study was to isolate cellulolytic and hemicellulolytic anaerobic bacteria inhabiting from gut of ruminants and investigate their hydrolytic enzyme activities. Extracellular CMCase activities of H-strains isolated from the rumen of a Holstein dairy cow were higher than those of D- and DC- strains from the rumen and large intestine of Korean spotted deer. Most isolated bacteria utilized more efficiently Dehority's artificial medium containing starch, glucose and cellobiose (DAS) than those in Dehority's artificial medium containing cellulose only (DAC). The results of biochemical reactions and sugar fermentation indicated that the isolated bacteria belong to one of bacterial strains of Peptostreptococcus spp., Bifidobacterium spp., Prevotela ruminicola/buccae, Clostridium beijer/butyricum and Streptococcus intermedis which are not highly cellulolytic. Activities of Avicelase, xylanase, β-D-glucosidase, α-L-arabinofuranosidase and β-xylosidase of the isolated anaerobic bacteria in DAS were higher than those in DAC. In conclusion, the results indicated the higher enzyme activities of the isolated strains cultured in DAS medium were mainly caused by their specific carbohydrate utilization for enzyme production and growth rate. The highly cellulolytic bacteria were not isolated in the present experiment. Thus further research is required to investigate characteristics of gut bacteria from Korean spotted deer.

Enzymatic Synthesis of β-Glucosylglycerol and Its Unnatural Glycosides Via β-Glycosidase and Amylosucrase

  • Jung, Dong-Hyun;Seo, Dong-Ho;Park, Ji-Hae;Kim, Myo-Jung;Baek, Nam-In;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.562-570
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    • 2019
  • ${\beta}$-Glucosylglycerol (${\beta}-GG$) and their derivatives have potential applications in food, cosmetics and the healthcare industry, including antitumor medications. In this study, ${\beta}-GG$ and its unnatural glycosides were synthesized through the transglycosylation of two enzymes, Sulfolobus shibatae ${\beta}$-glycosidase (SSG) and Deinococcus geothermalis amylosucrase (DGAS). SSG catalyzed a transglycosylation reaction with glycerol as an acceptor and cellobiose as a donor to produce 56% of ${\beta}-GGs$ [${\beta}$-$\text\tiny{D}$-glucopyranosyl-($1{\rightarrow}1/3$)-$\text\tiny{D}$-glycerol and ${\beta}$-$\text\tiny{D}$-glucopyranosyl-($1{\rightarrow}2$)-$\text\tiny{D}$-glycerol]. In the second transglycosylation reaction, ${\beta}$-$\text\tiny{D}$-glucopyranosyl-($1{\rightarrow}1/3$)-$\text\tiny{D}$-glycerol was used as acceptor molecules of the DGAS reaction. As a result, 61% of ${\alpha}$-$\text\tiny{D}$-glucopyranosyl-($1{\rightarrow}4$)-${\beta}$-$\text\tiny{D}$-glucopyranosyl-($1{\rightarrow}1/3$)-$\text\tiny{D}$-glycerol and 28% of ${\alpha}$-$\text\tiny{D}$-maltopyranosyl-($1{\rightarrow}4$)-${\beta}$-$\text\tiny{D}$-glucopyranosyl-($1{\rightarrow}1/3$)-$\text\tiny{D}$-glycerol were synthesized as unnatural glucosylglycerols. In conclusion, the combined enzymatic synthesis of the unnatural glycosides of ${\beta}-GG$ was established. The synthesis of these unnatural glycosides may provide an opportunity to discover new applications in the biotechnological industry.

Development of Detection Methods for Cellulolytic Activity of Auricularia auriculajudae

  • Jo, Woo-Sik;Bae, Soon-Hwa;Choi, Seung-Yong;Park, So-Deuk;Yoo, Young-Bok;Park, Seung-Chun
    • Mycobiology
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    • v.38 no.1
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    • pp.74-77
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    • 2010
  • To obtain basic information on the detection of cellulolytic activity in Auricularia auricula-judae, the influences of dye reagent, pH, and temperature were assessed. Chromogenic dye (congo red, phenol red, remazol brilliant blue, and trypan blue) was individually incorporated into a medium containing either carboxymethyl-cellulose, Avicel, or D-cellobiose as a polysaccharide carbon substrate. The other assessments utilized pHs ranging from 4.5 to 8.0 and temperatures from $15\sim35^{\circ}C$. Overall, when A. auricula-judae species were transferred onto media contained Congo red and adjusted pH 7.0 and then incubated at $25^{\circ}C$ for 5 days, the clear zone indicative of cellulolytic activity was more pronounced.