• Title/Summary/Keyword: enzymatic digest

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Modification of Acetobacter xylinum Bacterial Cellulose Using Dextransucrase and Alternansucrase

  • Kim, Do-Man;Kim, Young-Min;Park, Mi-Ran;Park, Don-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.704-708
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    • 1999
  • In addition to catalyzing the synthesis of glucan from sucrose as a primary reaction, glucansucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. Using dextransucrase and altemansucrase, prepared from Leuconostoc mesenteroides B-742CBM and B-1355C, respectively, we modified the bacterial cellulose in Acetobacter xylinum ATCC10821 culture, and then produced a characteristic cellulose that is soluble and has a new structure. There were also some partially modified insoluble cellulose and oligosaccharides in the modification culture. After methylation and following acid hydrolysis of both the soluble and insoluble glucans, there were ($1{\rightarrow}4$) as well as ($1{\rightarrow}6$) and ($1{\rightarrow}3$) glycosidic linkages in the soluble glucan.

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Five New Wood Decay Fungi (Polyporales and Hymenochaetales) in Korea

  • Kim, Nam Kyu;Park, Jae Young;Park, Myung Soo;Lee, Hyun;Cho, Hae Jin;Eimes, John A.;Kim, Changmu;Lim, Young Woon
    • Mycobiology
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    • v.44 no.3
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    • pp.146-154
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    • 2016
  • The wood decay fungi are a diverse taxonomic group that plays a pivotal role in forest carbon cycling. Wood decay fungi use various enzymatic pathways to digest dead or living wood in order to obtain carbon and other nutrients and these enzymatic systems have been exploited for both industrial and medical applications. Over 600 wood decay fungi species have been described in Korea; however, the recent application of molecular markers has dramatically altered the taxonomy of many of these wood decay fungi at both the genus and species levels. By combining molecular methods, specifically sequences of the internal transcribed spacer region, with traditional morphological characters, this study identified five new species records for Korea in five genera: Aurantiporus, Favolus, Neofavolus, Loweomyces, and Hymenochaetopsis. Three of these genera (Aurantiporus, Favolus, and Loweomyces) were previously unknown in Korea. The relatively simple morphology of the wood decay fungi often leads to ambiguous taxonomic assignment. Therefore, molecular markers are a necessary component of any taxonomic or evolutionary study of wood decay fungi. Our study highlights the need for a more robust and multifaceted approach in investigating new wood decay fungi in Korea.

Studies on Enzymes of the Higher Fungi of Korea( II ) - Identification of Cellulolytic Enzyme in Lenzites betulina- (한국산(韓國産) 고등(高等) 균류(菌類)의 효소(酵素)에 관한 연구(硏究)( II ) - 목재부후균(木材腐朽菌)인 조개껍질버섯의 섬유소 분해효소의 확인 -)

  • Park, Wan-Hee;Kim, Tae-Hee;Ro, Ihl-Hyeob
    • The Korean Journal of Mycology
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    • v.14 no.3
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    • pp.225-229
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    • 1986
  • Cellulosic substance which plays an important role in carbon cycle is most abundant in the nature world. Some higher fungi are able to digest cellulose directly to satisfy their carbohydrate requirement. Then, in order to investigate the enzymatic components of Lenzites betulina (Fr.) being wood rot, that fungus was collected in Kwangneung area. The carpophore of the fungus was smashed with cool distilled water, extracted and salted out by ammonium sulfate. And then the precipitate was purified by dialysing with visking tube and dissolved with pH 7.8 ammonia water, and the extract was filtrated. The fraction of filtrate was obtained as light brown powder after lyophilization, and determined cellulolytic activity. Cellulolytic potency of Lenzites betulina (Fr.) was 1. 65 unit/ml. The cellulase of Lenzites betulina (Fr.) was stable at below $45^{\circ}C$ and range of pH $4.5{\sim}6.0$ and is completely inactivated at $60^{\circ}C$ for 15 minutes. The optimum condition for the enzymatic reaction was $40^{\circ}C$ and pH 4.0. The enzyme activity was not influenced by the presence of $Ca^{2+}$ and $Fe^{2+}$.

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Enzymatic Modification of Cellulose Using Leuconostoc mesenteroides B-742CBM Dextransucrase

  • Kim, Do-Man;Kim, Young-Min;Park, Mi-Ran;Ryu, Hwa-Ja;Park, Don-Hee;Robyt, John F.
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.529-533
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    • 1999
  • In addition to catalyzing the synthesis of dextran from sucrose as a primary reaction, dextransucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. We have synthesized new glucans having new structures and new characteristics, by transferring D-glucose of sucrose to $\alpha$-cellulose and by using the constitutive dextransucrase obtained from Leuconostoc mesenteroides B-742CBM. The final reaction products were composed of soluble- and insoluble-glucans. The yields of soluble- and insoluble-glucans were theoretically 21% $\pm$ 2.2 and 68% $\pm$ 5.1, respectively. The remainder of the reaction products was recovered as a mixture of olgiosaccharides that could not be precipitated by 67%(v/v) ethanol. Treating the modified glucans with endo-dextranase and/or cellulase, oligosaccharides were produced that were not formed from the hydrolysis of native cellulose or B-742CBM dextran. The modification of the cellulose was confirmed by methylation and acid hydrolysis of the soluble-and insoluble-glucan. Both (1->4) and(1->6) glycosidic linkages were found in both of the glucans.

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Molecular Cloning and Expression of Bacillus pasteurii Urease Gene in Escherichia coli (B. pasteurii Urease 유전인자의 E. coli의 복제와 발현)

  • Kim, Sang-Dal;John Spizizen
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.297-302
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    • 1985
  • The 7.1 Mdal Xbaf fragment of Bacillus pasteurii ATCC 11859 containing gene for urease was inserted into the Xbal site of bifunctional plasmid pGR71, and its urease gene was cloned and expressed in E. coil RRI. But the cloned gene was not expressed in Bacillus subtilis BR151 in consequence of deletion of inserted DNA fragment. The recombinant plasmid thus formed was named pGU66. The restriction map of the plasmid pGU66 was determined, and the size of the plasmid was estimated to be 12.6 Mdal by double digestion of restriction enzymes of the plasmid. The urease of the cloned strain was accumulated in periplasmic space and very similiar to that of donor strains in their enzymatic properties.

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Anti-inflammatory activity of a sulfated polysaccharide isolated from an enzymatic digest of brown seaweed Sargassum horneri in RAW 264.7 cells

  • Sanjeewa, Kalu Kapuge Asanka;Fernando, Ilekkuttige Priyan Shanura;Kim, Eun-A;Ahn, Ginnae;Jee, Youngheun;Jeon, You-Jin
    • Nutrition Research and Practice
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    • v.11 no.1
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    • pp.3-10
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    • 2017
  • BACKGROUND/OBJECTIVES: Sargassum horneri is an edible brown alga that grows in the subtidal zone as an annual species along the coasts of South Korea, China, and Japan. Recently, an extreme amount of S. horneri moved into the coasts of Jeju Island from the east coast of China, which made huge economic and environmental loss to the Jeju Island. Thus, utilization of this biomass becomes a big issue with the local authorities. Therefore, the present study was performed to evaluate the anti-inflammatory potential of crude polysaccharides (CPs) extracted from S. horneri China strain in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. MATERIALS/METHODS: CPs were precipitated from S. horneri digests prepared by enzyme assistant extraction using four food-grade enzymes (AMG, Celluclast, Viscozyme, and Alcalase). The production levels of nitric oxide (NO) and pro-inflammatory cytokines, including tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-$1{\beta}$ were measured by Griess assay and enzyme-linked immunosorbent assay, respectively. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nuclear factor (NF)-${\kappa}B$, and mitogen-activated protein kinases (MAPKs) were measured by using western blot. The IR spectrums of the CPs were recorded using a fourier transform infrared spectroscopy (FT-IR) spectrometer. RESULTS: The polysaccharides from the Celluclast enzyme digest (CCP) showed the highest inhibition of NO production in LPS-stimulated RAW 264.7 cells ($IC_{50}$ value: $95.7{\mu}g/mL$). Also, CCP dose-dependently down-regulated the protein expression levels of iNOS and COX-2 as well as the production of inflammatory cytokines, including TNF-${\alpha}$ and IL-$1{\beta}$, compared to the only LPS-treated cells. In addition, CCP inhibited the activation of NF-${\kappa}B$ p50 and p65 and the phosphorylation of MAPKs, including p38 and extracellular signal-regulated kinase, in LPS-stimulated RAW 264.7 cells. Furthermore, FT-IR analysis showed that the FT-IR spectrum of CCP is similar to that of commercial fucoidan. CONCLUSIONS: Our results suggest that CCP has anti-inflammatory activities and is a potential candidate for the formulation of a functional food ingredient or/and drug to treat inflammatory diseases.

Optimization of Culture Conditions for the Production of Diphtheria Toxin (디프테리아 toxin 생산을 위한 발효조건 최적화)

  • Cho, Min;Ryu, Yeon-Woo
    • KSBB Journal
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    • v.14 no.2
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    • pp.241-247
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    • 1999
  • Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

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ISOLATION AND IDENTIFICATION OF ANAEROBIC RUMEN BACTERIUM, ACTINOMYCES SP. 40 AND ENZYMATIC PROPERTIES OF β-1, 4-ENDOGLUCANASE

  • Min, H.K.;Choi, Y.J.;Ha, J.K.;Cho, K.K.;Kwon, Y.M.;Chang, Y.H.;Lee, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.373-382
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    • 1994
  • A bacterial strain No. 40, which produced extracellular endoglucanase, was isolated from the rumen of Korean native goals and identified to be a genus of Actinomyces sp. The optimum conditions for endoglucanase production in PY-CMC medium were initial pH of 7.0 and 4 days of cultivation at $39^{\circ}C$. When localization of endoglucanase activity of Actinomyces sp. was determined, 68% of the enzyme activity was found in the extracellular fraction, 11% of the activity was detected in the periplasmic space and the remaining activity was in the intracellular and cell-bound fractions. The maximal endoglucanase activity was observed at pH 5.0 and it was most s table at pH 5.0. The optimum temperature of this enzyme activity was $55^{\circ}C$, but enzyme activity was gradually lost at temperature above $60^{\circ}C$. The crude enzyme was activated by addition of 10 mM cysteine and 10 mM DTT. But it was inhibited by addition of 10 mM $Cu^{{+}{+}}$ and $Fe^{{+}{+}}$. This crude enzyme could digest carboxymethylcellulose (CMC), and degrade xylan, avicel, pNPG, and pNPC to a less extent.

BIOACTIVE PEPTIDES DERIVED FROM FOOD PROTEINS AND PREVENTION OF LIFE-STYLE RELATED DISEASES

  • Yoshikawa Masaaki
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2001.12a
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    • pp.69-73
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    • 2001
  • Two opioid peptides, YPLDL and YPLDLF, were isolated from enzymatic digests of spinach ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO) and named rubiscolin-5 and -6, respectively. These peptides were selective for delta-receptor and the latter was about 3 times more potent than the former. After oral administration in mice at the dose of 100 mg/kg, rubiscolin-6 showed analgesic activity in tail pinch test. It also stimutated learning performance at the same dose in passive avoidance experiment using step-through apparatus. An immunostimulating peptide, MITLAIPVNKPGR, was isolated from a trypsin digest of soybean protein and named soymetide. Immunostimulating activy of soymetide was mediated by fMLP receptor. Interestingly, after oral administration in rats at a dose of 300 mg/kg (po.), soymetide-4 (MITL) protected alopecia (hair-loss) induced by etoposide, a cancer chemotherapy agent. Stimulation of IL-1 release by the peptide was involved in the mechanism. Ovokinin(2-7), RADHPF, is a vasorelaxing peptide released from ovalbumin by the action of chymotrypsin. It lowered blood pressure of spontaneously hypersensive rats (SHR) after oral administration at a dose of 10 mg/kg. RPLKPW, which was designed by replacing 4 amino acid residues in ovokinin(2-7), exhibited hypotensive activity at a dose of 0.1 mg/kg (po.). This peptides was introduced into 3 homologous sites in soybean beta-conglycinin alpha' subunit by site-directed mutagenesis of the cDNA and expressed in E. coli. The minimum effective dose for hypotensive activity of the genetically modified beta-conglycinin alpha' subunit was 10 mg/kg (po.), which is about 1/200 that of ovalbumin.

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Enzymatic Synthesis of New Oligosaccharides Using Glucansucrases. (Glucansucrases를 이용한 새로운 올리고당의 합성)

  • ;;;;;John F. Robyt
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.179-186
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    • 1998
  • Dextransucrase hyper-producing Leuconostoc mesenteroides B-512FMCM and dextransucrase constitutive mutants B-742CB and B-1355C catalyzed the transfer of glucose from sucrose to other carbohydrates which were present or were added to the reaction digests. When the acceptor was a maltose, gentiobiose, lactose or raffinose, there was produced a series of oligosaccharide acceptor products or single product based on the kinds of enzymes and reaction conditions. To obtain the quantitative information about the yield and the distribution of acceptor products and dextran two experimental parameters were studied: a) the ratio of acceptor to sucrose and b) the amount of enzyme at constant carbohydrate concentration (100 mM). As the amount of enzyme increased, the synthesis of acceptor products (of maltose or gentiobiose) increased, and the formation of dextran decreased. As the ratio of acceptor to sucrose increased, the amount of dextran and the number of acceptor-products decreased and the amount of acceptor-products increased. When maltose or gentiobiose was an acceptor, the glucose from sucrose was transferred to the C-6 hydroxyl group of the nonreducing-end glucose residue of accepters to give a homologous series of isomaltosyl dextrins. In case of lactose or raffinose, there was produced only one acceptor product from B-512FMCM dextransucrase reaction. In the lactose acceptor reaction, the glucose from sucrose was transferred to the C-2 hydroxyl of the reducing end glucose residue of lactose. To get a series of oligosaccharides from lactose or raffinose acceptor reaction we used B-742CB dextransucrase or B-1355C alternansucrase with 500 mM sucrose in reaction digest.

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