• Title/Summary/Keyword: cellobiose

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Effect of Xylan on Production of Xylanolytic Activity from Penicillium verruculosum (Penicillium verruculosum의 Xylan분해활성도의 생성에 대한 Xylan의 영향)

  • 조남철;정두례;유영균
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.4
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    • pp.423-427
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    • 1992
  • During the cultivation of Penicillium verruculosum in the medium containing xylan as a sole carbon source for 26 days, xylanolytic activity and some changes were investigated. Protein content and xylanolytic activity, p-Nitrophenyl-$\beta$-D-xylopyranoside (PNPX), p-Nitrophenyl-$\beta$ -D-glucopyranoside (PNPG) hydrolytic activities were increased until 8 days but reducing sugar content was not correlated to protein content. When crude proteins from the culture broth were separated on SDS-PAGE, distribution of proteins was different from the culture broth of cellobiose octaacetate (COA) medium. The culture broth of xylan medium had high hydrolytic activity on xylan but not on cellulose. Furthermore, xylanolytic products were showed xylose, xylobiose and oligosaccharides on thin layer chromatography, and xylobiose was major product. Those result suggested that xylanolytic activity of culture broth was endo-type hydrolysis. Optimum temperatures of xylanolytic activity and PNPX hydrolytic activity of culture broth were 50~6$0^{\circ}C$ and 60~7$0^{\circ}C$, respectively and optimum pHs were 3.0~4.0 and 4.0~5.0, respectively.

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Characterization of Fusarium oxysporum Isolated from Paprika in Korea

  • Cha, Sang-Do;Jeon, Young-Jae;Ahn, Geum-Ran;Han, Jae-In;Han, Kap-Hoon;Kim, Seong-Hwan
    • Mycobiology
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    • v.35 no.2
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    • pp.91-96
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    • 2007
  • In the present study we first report in Korea the identification and characterization of Fusarium oxysporum isolated from rotten stems and roots of paprika (Capsicum annuum var. grossum) at Masan, Kyungsangnamdo in 2006. The fungal species produced white aerial mycelia accompanying with dark violet pigment on PDA. The optimal temperature and pH for the growth of the species was $25^{\circ}C$ and pH 7, respectively. Microscopic observation of one of isolates of the species shows that its conidiophores are unbranched and monophialides, its microconidia have oval-ellipsoidal shape with no septate and are of $3.0{\sim}11{\times}1.5{\sim}3.5\;{\mu}m$ sizes, its macroconidia are of $15{\sim}20{\times}2.0{\sim}3.5\;{\mu}m$ sizes and have slightly curved or slender shape with $2{\sim}3$ septate. The results of molecular analysis show that the ITS rDNA of F. oxysporum from paprika shares 100% sequence identity with that of known F. oxysporum isolates. The identified species proved it's pathogenicity by causing rotting symptom when it was inoculated on paprika fruits. The growth of F. oxysporum from paprika was suppressed on PDA by agrochemicals such as benomyl, tebuconazole and azoxystrobin. The identified species has the ability of producing extracelluar enzymes that degrade cellobiose and pectin.

Crystal Structure of Hypothetical Fructose-Specific EIIB from Escherichia coli

  • Park, Jimin;Kim, Mi-Sun;Joo, Keehyung;Jhon, Gil-Ja;Berry, Edward A.;Lee, Jooyoung;Shin, Dong Hae
    • Molecules and Cells
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    • v.39 no.6
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    • pp.495-500
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    • 2016
  • We have solved the crystal structure of a predicted fructose-specific enzyme $IIB^{fruc}$ from Escherichia coli ($EcEIIB^{fruc}$) involved in the phosphoenolpyruvate-carbohydrate phosphotransferase system transferring carbohydrates across the cytoplasmic membrane. $EcEIIB^{fruc}$ belongs to a sequence family with more than 5,000 sequence homologues with 25-99% amino-acid sequence identity. It reveals a conventional Rossmann-like ${\alpha}-{\beta}-{\alpha}$ sandwich fold with a unique ${\beta}$-sheet topology. Its C-terminus is longer than its closest relatives and forms an additional ${\beta}$-strand whereas the shorter C-terminus is random coil in the relatives. Interestingly, its core structure is similar to that of enzyme $IIB^{cellobiose}$ from E. coli ($EcIIB^{cel}$) transferring a phosphate moiety. In the active site of the closest $EcEIIB^{fruc}$ homologues, a unique motif CXXGXAHT comprising a P-loop like architecture including a histidine residue is found. The conserved cysteine on this loop may be deprotonated to act as a nucleophile similar to that of $EcIIB^{cel}$. The conserved histidine residue is presumed to bind the negatively charged phosphate. Therefore, we propose that the catalytic mechanism of $EcEIIB^{fruc}$ is similar to that of $EcIIB^{cel}$ transferring phosphoryl moiety to a specific carbohydrate.

Design of Recycle Bubble Column Reactor for Continuous Enzymatic Hydrolysis of Cellulose (섬유소의 연속 효소 가수분해를 위한 순환식 기포탑 반응기의 설계)

  • 김춘영;홍석표정봉우이태원
    • KSBB Journal
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    • v.5 no.1
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    • pp.59-67
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    • 1990
  • Enzymatic hydrolysis of insoluble cellulose was performed in a bubble column with tangential flow ulrafiltration membrane unit. The reactor was operated in a batch mode as well as semi-continuous and continuous with continuous removal of products through the tangential flow ultrafiltration membrane. The optimum superficial gas velocity was 1-3cm / sec so as to avoid bubble coalescence and enzyme denaturation. In continuous and selni-cotinuous process, the conversion was gradually increased but the total reduced sugar concentration was drcastically dereased with the dilution rate. It was concluded that the bubble column attaching tangential flow ultrafiltration membrane unit was effective on continuous hydrolysis of cellulose and recovery of enzyme.

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Characterization of Carboxymethylcellulase(CMCase) Produced by Recombinant E. coli Containing CMCase Gene for Cellulomonas sp. YE-5

  • Park, Sung-Won;Her, Nam-Yun;Kim, Dong-Seob;Park, Sun-Jin;Lee, Han-Seung;Park, Hak-Jong;Yu, Ju-Hyun
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.174-179
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    • 1997
  • CMCase produced by recombinant E. coli JM109 (pCEH#4) containing CMCase gene from Cellulomonas sp. YE-5 was purified to 24.3 fold and 2.6% yield by ammoniumsulfate precipitation, DEAE-cellulose column chromatography and gel filtration on Sephadex G-100. The optimum pH and temperature for CMCase activity were pH 7.0 and 5$0^{\circ}C$. The enzyme was stable between pH 5.0 and 10.0, and up to 6$0^{\circ}C$. The molecular weight of he enzyme was estimated to be approximately 40,000 daltons by SDS-PAGE. Analysis of the amino acid composition showed that the enzyme contained many glycines and acidic amino acids. The enzyme was an endo-type CMCase and the final enzyme reaction product from hydrolysis of Cm-cellulose by the enzyme was cellobiose. {TEX}$K_{M}${/TEX} value determined with CM-cellulose was 1.28mM.

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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.

Isolation and Characterization of a New Cellulase-producing Marine Bacterium, Seonamhaeicola sp. S2-3 (셀룰로스분해 신규 해양미생물 Seonamhaeicola sp. S2-3의 분리 및 동정)

  • Kim, Da Som;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.539-546
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    • 2020
  • A cellulolytic bacterial strain, S2-3, was isolated from sea water collected in Jeju island, Republic of Korea. The strain was aerobic and gram negative, and formed yellow colored colonies on marine agar medium. S2-3 cells were long rod-shaped, 0.5 × 0.25 ㎛ (width x length) in size, and did not have flagella. The optimal growth conditions for S2-3 were 30-35℃ and pH 6.5-7.0. Analysis of the 16S rRNA gene sequence of S2-3 revealed that it had the highest identity with those of Seonamhaeicola algicola Gy8 (97.08%), Hyunsoonleella udonensis JG48 (95.01%), and Aestuariibaculum scopimerae I-15 (94.86%). In phylogenetic analysis, S2-3 formed the same clade as S. algicola Gy8, implying that S2-3 belongs to the genus Seonamhaeicola. The major fatty acids (>10%) comprised C15:1 iso G (22.29%), C15:0 iso (17.71%), C17:0 iso 3OH (16.06%), and C15:0 iso 3OH (10.7%), resulting in quite different ratio of the component from those of S. algicola Gy8. Moreover, its biochemical characteristics, including acid production and enzyme activities, were different from those of S. algicola Gy8. Therefore, putting all these results together, we concluded S2-3 is distinct species from S. algicola Gy8, and thus named it Seonamhaeicola sp. S2-3. In liquid culture, S2-3 produced extracellular cellulases that can hydrolyze cellulose or cellooligosaccharides into cellobiose, which is a good enzyme resource that deserves further research.

Effect of the Level of Carbohydrates on Bio-hydrogenation and CLA Production by Rumen Bacteria When Incubated with Soybean Oil or Flaxseed Oil In vitro (Soybean Oil 및 Flaxseed Oil 첨가 배양시 탄수화물 첨가수준에 의한 반추미생물의 Bio-hydrogenation과 CLA 생성에 미치는 효과)

  • 최성호;임근우;김광림;송만강
    • Journal of Animal Science and Technology
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    • v.48 no.4
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    • pp.521-532
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    • 2006
  • An in vitro study was conducted to examine the effect of addition level of carbohydrates on fermentation characteristics, and bio-hydrogenation of unsaturated fatty acids by mixed rumen bacteria when incubated with soybean oil or flaxseed oil. Four levels(0%, 0.3%, 0.6% and 0.9%, w/v) of the mixed carbohydrates(glucose, cellobiose, soluble starch, 1:1:1, in weight basis) and oil sources(soybean or flaxseed oil, 60mg in 150ml culture solution) were added to the mixed solution of strained rumen fluid with artificial saliva(1:4, v/v), and incubated anaerobically for 12 hours at 39℃. pH and ammonia-N concentration were lower by increasing the substrate levels at all incubation periods(P<0.05~P<0.001). The propionate proportion increased(P<0.001), but acetic acid and butyric acid decreased(P<0.001) with the substrate level at 6 and 12 h incubations. Oil sources did not influence the proportions of individual VFA. At the end of incubation, the proportions of C18:0(P<0.01), C18:1(P<0.001) and trans-11C-18:1(P<0.01) were reduced but those of C18:2(P<0.001) and C18:3(P<0.01) were enhanced by the addition of flaxseed oil compared to addition of soybean oil. The proportions of C18:0 and total CLA were reduced(P<0.01) but those of trans-11-C18: (P<0.05) and C18:2(P<0.01) were increased with the substrate level when incubated with soybean oil or flaxseed oil. There were interactions(P<0.05) in the proportions of C18:1, C18:2 and C18:3(P<0.01) between oil source and substrate level. The proportions of cis-9, trans-11-CLA and trans-10, cis-12-CLA tended to reduce with substrate level, although there was no significant difference between treatments.

Red-Colored Phenomena and Morphochemical Characteristics of Red-Colored Substances in Ginseng Roots (Panax ginseng C.A. Meyer) (인삼 적변현상과 적변물질의 형태-화학적 특성)

  • 윤길영;양덕조
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.107-112
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    • 2000
  • One of the physiologically important ginseng diseases is red-colored phenomena (RCP) that is caused by accumulation of red-colored substances on the epidermis of ginseng roots. Although RCP severely deteriorates the quality of ginseng products, there has been little information on what red-colored substance is and how RCP occurs. Therefore, the heavy losses of cultivators and ginseng industry are suffering by RCP, For this reason, we have investigated with the morphochernical characteristics of RCP to find out main cause of it. The red-colored substances (RS) on the epidermis of red-colored ginseng (RCG) were examined using inverted light microscope, confocal laser scanning microscope (CLSM)and furier transform infrared (FT/IR) spectrometer. Red brown substances were accumulated in the cell wall of the epidermis from early stage to late stage of RCC. Especially, cell wall of the late stage of RCG was covered with the sub-stances with 80~ 130 fm thick. Therefore, the cell wall of RCG cannot protect the ginseng root cells from the mechanical damages, bacteria and fungi. To analyse red substances of roots, RS were isolated from epidermis of RCG and extracted using various solvents. RS is strongly insoluble but it was bleached by oxidizing agents including 12% (v/v) NaOCl. Therefore, RS was Presumed to make up of high chelation power. The proriles of FT/IR spectra or both healthy ginseng (HEG) and RCG showed a significant difference at two wavelength,2857 cm$\^$-1/(C-H) and 1032 cm$\^$-1/(S=O), respectively. Furthermore, absorption peak of 2857cm$\^$-l/ appears on the only epidermis of RCG. The other peak is shown lower absorption rate on the epidermis of RCG than that of healthy ginseng. Also, FT/IR spectra of the mixture of carboxym-ethylcellulose (CMC) and iron (Fe$\^$3+/) were very similar to RCG spectrum profiles. One of a interesting fact is that the contents of phenolic compounds at the epidermis of healthy ginseng were highest. The results of these experiments sup-port the RCP was closely related with the chemical interaction between inorganic elements (Fe) of rhizosphere and organic matters (cellulose, cellobiose, cell sap, etc.) of ginseng roots.

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Cultural Characteristics of Fomitella fraxinea (Fr.) Imaz. (장수버섯의 배양적(培養的) 특성(特性))

  • Chang, Hyun-You;Cha, Dong-Yeul;Kang, An-Seok;Hong, In-Pyo;Kim, Kwang-Po;Seok, Soon-Ja;Ryu, Young-Jin;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.23 no.3 s.74
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    • pp.238-245
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    • 1995
  • A good mycelial growth of F. fraxinea was observed on CDA medium. The optimum temperature and pH for the mycelial growth of F. fraxinea was at $30^{\circ}C$ and pH 6.0, respectively. Glucose was found to be the best carbon source and arginine was favored as nitrogen source. When the basal medium was supplemented with organic acids, the best growth was shown in succinic acid and the poor growth was shown in oxalic acid. Thiamine.HCl showed the best results on the growth of this fungus on basal medium. Mycelial growth of F. fraxinea was quite good when oak tree sawdust was used to cultural substrates. The best mycelial growth was observed when 20% of rice bran was added as a supplement on sawdust substrates. Higher yield of F. fraxinea was observed on the medium with oak tree and acacia sawdust.

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