• Title/Summary/Keyword: formation of maltose

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Current Studies on Browning Reaction Products and Acidic Polysaccharide in Korean Red Ginseng (홍삼에 함유된 갈변물질 및 산성다당체에 대한 연구현황)

  • Lee, Jong-Won;Do, Jae-Ho
    • Journal of Ginseng Research
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    • v.30 no.1
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    • pp.41-48
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    • 2006
  • In the browning reaction of Korean ginseng, it appears that enzymatic and non-enzymatic browning reaction occurred In initial stage of steaming fresh ginseng at low temperature, and then non-enzymatic browning reaction followed in the drying period after steaming. Browning reaction of red ginseng occurred between $60{\sim}90$ min of steaming at $100^{\circ}C$, and browning pigments of red ginseng were mostly water soluble substances. The structural characteristics of water soluble browning reaction products(WS-BRPs) isolated from Korean red ginseng were showed the presence of hydroxyl, amide carbonyl and aliphatic methane groups. From sugar analysis it was identified that L and S-1, melanoidins isolated from red ginseng, contained two kinds of sugars, glucose and xylose, and the other melanoidin S-2 contained the previous and fructose. In order to find out pertinent methods for the acceleration of browning during ginseng processing, various treatment were made on fresh ginseng with sugars, amino acids and inorganic nitrogenous compounds and the extent of browning was measured. Among sugar tested, maltose resulted in the greatest acceleration of browning followed in decreasing order by glucose and lactose, whereas pentoses, fructose, sucrose and raffinose had negligible effect. A marked browning occurred in ginseng treated with basic amino acids, while the extent of browning was not greatly increased when ginseng was treated with aliphatic amino acids, hydroxyl amino acids, or acidic amino acids. The brown color intensity gradually increased with an increase of glucose concentration far up to 0.5M. L, S-1, and S-2 were found to have an ability to donate hydrogen to DPPH, and also they had anti-oxidative activity in the experiments of hydrogen peroxide scavenging, inhibitory activity in the formation of MDA from linoleic acid, auto oxidation of ok-brain homogenates, lipid peroxidation by the enzymatic and non-enzymatic system in liver microsome fraction, and mitochondrial fraction etc. The amounts of acidic polysaccharide(AP) in red ginseng were higher than those of wild and cultured Panax quinquefolius, Panax notoginseng as well as white ginseng (Panax ginseng). In white ginseng, the AP amount is no difference in root ages or sizes, also, the AP amount of ginseng body was similar to that of rhizome, but was higher than that of leaf and epidermis. Addition of red ginseng acidic polysaccharide(RGAP) increased production of nitric oxide(NO) and tumor necrosis factor (TNF)-$\alpha$ in the rodent macrophage cultures, and treatment of RGAP in vivo stimulated tumoricidal activities of natural killer (NK) cells.

Improvement of Transglycosylation Efficiency using a Glycosynthase Mutant derived from Thermoplasma acidophilum ${\alpha}$-Glucosidase (Thermoplasma acidophilum 유래 ${\alpha}$-glucosidase로 부터 생산된 glycosynthase 돌연변이 단백질의 개선된 당전이 효율)

  • Hwang, Sung-Min;Seo, Seong-Hwa;Park, In-Myoung;Choi, Kyoung-Hwa;Kim, Do-Man;Cha, Jae-Ho
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.104-110
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    • 2012
  • Glycosynthase is an active site nucleophile mutant enzyme, prepared from glycosidase, which is capable of synthesizing oligosaccharide derivatives without the hydrolysis of the product. Thermoacidophilic ${\alpha}$-glucosidase of Thermoplasma acidophilum (AglA) exhibits a transglycosylating activity yielding various glycosides. AglA was converted to glycosynthase by the substitution of the catalytic nucleophile Asp-408 residue into non-nucleophile glycine in order to increase its ability to synthesize various glycosides by transglycosylation. The glycosynthase mutant was purified by Ni-NTA chromatography and its glycoside-synthesizing activity was measured by using an external nucleophile, sodium formate buffer, providing maltose as a donor and p-nitrophenyl-${\alpha}$-D-glucopyranoside ($pNP{\alpha}G$) as an acceptor, respectively. In addition, $pNP{\alpha}G$ was examined for its feasibility to act as both a donor and an acceptor, and products were compared with those of the wildtype enzyme. The mutant enzyme was found to catalyze the formation of a specific product from $pNP{\alpha}G$ with a yield of 42.5% without further hydrolysis, while the wild-type enzyme produced two $pNP{\alpha}G$ products at low yields. The results demonstrate the possibility of satisfactory yields for the reactions in the presence of small amounts of acceptor, and demonstrate that the high activity of the mutant, at pHs below neutrality, was applicable in the transfer of glucose from the natural donor.

Plant Regeneration from Mature Seed-Derived Callus in Bermudagrass (Cynodon dactylon) (난지형 목초 버뮤다그라스의 종자유래 캘러스로부터 식물체 재분화)

  • Lee, Ki-Won;Park, Hyung-Soo;Choi, Gi-Jun;Kim, Ki-Yong;Ji, Hee-Chung;Kim, Kyung-Hee;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.3
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    • pp.223-228
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    • 2011
  • The present study was conducted to determine the optimum in vitro culture condition for callus induction and plant regeneration from mature seeds of bermudagrass (Cynodon dactylon cv. Common). It was revealed that mature seeds cultured on MS medium supplemented with 2 mg/L 2,4-D, 0.5 g/L proline, 0.5 g/L casamino acid and 3 g/L Gelrite under light condition produced the highest percentage of callus formation (39.2%). The most suitable medium for plant regeneration from dehydrated calli was MS agar medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L BA, 0.5 g/L proline, 0.5 g/L casamino acid 3 g/L Gelrite which induced the highest percentage of calli forming shoots (57.7%). The frequency of callus induction and plant regeneration were the highest on sucrose, followed by maltose. The shoots were rooted at the highest rate (100%) when transferred onto 1/2 MS medium. Regenerated plants were morphologically uniform with normal growth pattern.

Isolation and Characterization of Cryptococcus sp. CS-2 Secreting Polygalacturonase from Soil (토양으로부터 Cryptococcus sp. CS-2의 분리 및 균주가 분비하는 Polygalacturonase의 특성에 관한 연구)

  • 강희경;문명님;임채영;양영기
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.158-163
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    • 1999
  • A ploygalacturonase-produchg yeast was isolated from Cheju soil by selective eivichment media. One strain which has the highesl activity of polygalacturonase was selected. The characle~ishcs of the strain CS-2 were as follows: CS-2 utilized xylose. sucrose, maltose, u.ehalose, cellobiose. melibiose, lactose, raffinose, inosiiol, dulicilol, and dextrose, but did not utilized galactose, nitrate. nit~te, and lysine. Growth of CS-2 was inhibited by cyclohexamide, 1% acetic acid, and high concenaation (over 50%) of glucose. It grew at $30^{\circ}C$ but did 'IIOL $35^{\circ}C$. The cell size ofthe strain CS-2 was 2.9 p ~ n in length and 1.3 $\mu$ in diameter. Vegetable reproductmn was multiple budding and ascospre was present I to 4. Pseudomycelia or true myceliua formation were not observed In any of the cullureq. These results suggest that strain CS-2 is most likely a strain related Cryptococcus spp. (Cryptococcu spp. CS-2). When polygalacturonase or ihe yeast was induced by addition of polygalactoronic acid, polygalacturonase activity was detected in culture supernatent. There was a peak of specific activity a1 he mid-stationary phase(3 days culture) of growth. Polygalacturonase specific activity of Crylmcoccus sp. CS-2 was 2.96 unitsling. The molecular weighl ol'polygalacturonase was showed to be 46 KDa by both SDS-PAGE and activity stailling.

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Studies on the Ecology of Occurrence and Identification of Typhula Snow Mold of Graminous Plants -II. Several Factors Affecting Growth of Typhula incarnata- (화본과식물에 발생하는 설부소입균핵병균(雪腐小粒菌核病菌)의 동정 및 발생상태에 관한 연구 -II. Typhula incarnata의 생육에 미치는 몇 가지 요인-)

  • Kim, Jin-Won;Lee, Du-Hyung;Shim, Gyu-Yul
    • The Korean Journal of Mycology
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    • v.20 no.1
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    • pp.37-43
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    • 1992
  • Typhula incarnata grew over a temperature range of -5 to $20^{\circ}C$ with maximum growth at 10 to $15^{\circ}C$. Sclerotial production for T. incarnata was greatest at the higher temperature. Maximum mycelial growth of this pathogen occurred from pH 5.4 to 6.2. When carbon sources were added to a basal salt medium (Czapek's dox agar) at 5 g carbon sources/l, inulin, soluble starch, galactose, glucose, mannose, manitol, sucrose, maltose, cellobirose, trehalose, raffinose, and dextrin supported growth better than other carbon sources did. Of the twenty-three nitrogen sources tested, glycine, serine, ammonium sulfate, asparagine, asparatic acid, and ${\beta}-alanine$ were the most favorable for mycelial growth of T. incarnata. Cystine and cysteine were poor nitrogen sources. Ammonium salt of nitrogen sources supported growth better than nitrate salt of nitrogen sources. Potato dextrose agar, oat meal agar, and V-8 juice agar were the most favorable for mycelial growth and sclerotial formation. Appropriate addition of pepton to PDA decreased mycelial dry weight, but sucrose supported good growth of T. incarnata. Percent viable sclerotia of T. incarnate buried in bentgrass soil decreased from 2 months after treatment remarkably. Trichoderma riride and bacteria were isolated from non-germinated sclerotia. Live orchard grass leaf pieces within the soil were colonized by T. incarnata better than sterile and unsterile dead leaf pieces at $0^{\circ}C$. Saprophytic ability of T. incarnate on sterile leaf sheath occurred better at $0^{\circ}C$ than at $10^{\circ}C$. Saprophytic microflora consisting of Cladosporium sp., Fusarium sp., Mucor sp., Pythium sp., and unidentified fungi were the competitors for the sterilized and unsterilized substrate, but their colonization was not find on live leaf sheath buried in the soil at $0^{\circ}C$. In the effect of fungicides to Typhula snow mold disease of creeping bentgrass, mixture of polyoxin and thiram was the most effective, followed by iprodione, mixture of iprodione and oxine copper, thiophanate-methyl, myclobutanil, and tolclofos-methyl.

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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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