• Title/Summary/Keyword: hansenula sp.

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Dicyma sp. YCH-37이 생산하는 효모세포벽 용해효소 I. 생산균주의 분리 및 효소의 정제

  • Chung, Hee-Chul;Hahm, Byoung-Kwon;Bai, Dong-Hoon;Hasegawa, Toru;Yu, Ju-Hyun
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.445-451
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    • 1996
  • The strain YCH-37, which produces yeast cell wall lytlc enzyme, was isolated from soil. From the microscopic observation, morphological and cultural characteristics, this strain was identified to fungus, Dicyma sp. So, we named this strain as Dicyma sp. YCH-37. The lytic enzyme effectively lysed Salmonella typhimurium among intact living bacteria and Torulopsis, Hansenula, Zygosaccharomyces among intact living yeast, as well as autoclaved yeast strains. The yeast cell wall lytic enzyme was succesively purified to 204 folds with 13% yields through yeast glucan affinity adsorption and DEAE-cellulose column chromatography. The enzyme was identified to monomeric protein with molecular weight of 25,000 daltons from the results of SDS-PAGE and gel filtration. The optimum pH and temperature for the yeast lytic activity were 8.0 and 50$\circ$C, respectively. The enzyme was stable up to 40$\circ$C, and between pH 4.0-pH 10.0.

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Studies on the Yeast-Like Fungi Associated with Bovine Mastitis 2. Sensitivity of Yeast-Like Fungi to Antifungal Agents (유우유방염(乳牛乳房炎)에 관여(關與)하는 효모양진균(酵母樣眞菌)에 관한 연구(硏究) 2. 효모양진균(酵母樣眞菌)의 항진균성물질(抗眞菌性物質)에 대한 감수성(感受性))

  • Yeo, Sang-Geon;Choi, Won-Pil
    • Korean Journal of Veterinary Research
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    • v.22 no.2
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    • pp.139-147
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    • 1982
  • A total of 133 isolates of yeast-like fungi was tested for sensitivity to seven different antifungal agents. The yeast-like fungi tested were isolated from the milk from normal or mastitic bovine quaters or from bovine feces. They were 5 Candida albicans (C. albicans) isolates, 63 C. krusei, 27 C. tropicalis, 5 C. parapsilosis, 10 Torulopsis glabrata, 6 Rhodotorula sp., 6 Hansenula sp. and 1 Pichia sp. isolate. The antifungal agents tested were nystatin, griseofulvin, cycloheximide, 5-fluorocytosine, miconazol, clotrimazole and tolnaftate. In general, clotrimazole, miconazol and 5-fluorocytosine were more effective in antifungal activity in vitro against the test organisms than the rest of the agents tested. However, some of the isolates showed higher resistance to certain antifungal agents compared to the other isolates of the some species. They were: 1 C. albicans isolate to 5-fluorocytosine; 1 C. albicans to 5-fluorocytosine, miconazol and clotrimazole; 1 C. krusei to 5-fluorocytosine and cycloheximide; and 11 C. tropicalis isolates to cycloheximide. The minimum inhibitory concentrations(MIC) of clotrimazole were $12.5{\mu}g/ml$ or lower for all isolates tested except one C. albicans isolate, for which MIC of the drug was $100{\mu}g/ml$. On the other hand, the MIC's of cycloheximide were $6.5{\mu}g/ml$ or lower for all isolates except the following; all isolates of C. albicans ($100{\mu}g/ml$), C. pseudotropicalis ($200{\mu}g/ml$) and Rhodotorula sp. ($25-50{\mu}g/ml$), 11 C. tropicalis isolates ($100{\mu}g/ml$) and 1 C. krusei isolate ($200{\mu}g/ml$).

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Expression of heterologous genes using the slpA promoter and signal sequence in Lactobacilli

  • Gang, Seung-Ha;Bok, Jin-Deok;Jo, Gwang-Geun;Jo, Jae-Sun;Choe, Yun-Jae
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.202-205
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    • 2000
  • A gene coding $endo-{\beta}$,-1, 4 glucanase from Actinomyces sp. KNG40 and phytase from Hansenula polymorpha were cloned into Esherichia coli JM101 by using E. coli/Lactobacillus shuttle vector pNZ3004 and pNZ123. The plasmid p3PS(1-4) and p123(1-4) have slpA promoter and slpA signal sequence. So, I constructed expression vectors, p3PS(1-4)Endo, phy and p123(1-4)Endo, phy. These constructed vector was transformed in target host Lactobacillus gasseri and reutri. These transformed host expressed endoglucanase and phytase as extracellular fraction. In the enzyme activity of the same vector, host L, gasseri was higher activity than L. reuteri. This indicates that L. gasseri recongnize promoter and signal sequence very well.

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Changes in microorganisms and Min Components during Takju Brewing by a Modified nuruk (개량누룩에 의한 탁주양조중 미생물과 중요성분 변화)

  • 이명숙
    • The Korean Journal of Food And Nutrition
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    • v.12 no.3
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    • pp.226-232
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    • 1999
  • To evaluate the characteristics of modified nuruk made by inoculation and cultivation of Rhizopus jap-onicus T2, Aspergillus oryzae L2 and hansenula sp. BC26 three different Takju mashes were made with modified Nuruk commercial Nuruk and rice koji and the changes in microorganisms and major compon-ents of mashes were investigated uring brewing. The numbers of yeast kept higher in the mash of mod-ified Nuruk or rice koji. The numbers of lactic acid bacteria were high in mash of commercial Nuruk but those were not in mash of modified Nuruk or rice koji. all mashes showed stable pH in a day and the pH kept higher in mash of modified Nuruk than in that of commer-cial Nuruk or rice koji. Increase in the content of total acid was moderate in mash of modified Nuruk or rice koji but it was too much in that of commercial Nuruk. increase in the content of amino acid was much highr in mash of modified Nuruk or commercial Nuruk than in that of rice koji. Increase in the content of alcohol was more rapid in mash of modified nuruk or rice koji than in that of commercial Nur-uk. The content of reducing sugar kept highter in mash of commercial Nuruk or modified Nuruk than in that of rice koji. A small amount of modified Nuruk accomplished favorable fermentation showing nor-mal patterns in microbiological and physicochemical changes during brewing.

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Characteristics of a Modified Nuruk Made by Inoculation of Traditional Nuruk Microorganisms (전통 누룩미생물들로 제조한 개량누룩의 특성)

  • 소명환
    • The Korean Journal of Food And Nutrition
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    • v.12 no.3
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    • pp.219-225
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    • 1999
  • In order to improve the quality of nuruk a Korean traditional fermenting agent for brewing nuruk was modified by a new method inoculating Rhizopus japonicus T2 Aspergillus oryzae L2 and hansenula sp. BC26 which had been isolated from traditional Nuruk. And the characteristics of modified Nuruk were investimgated as compared with current fermenting agents such as commercial Nuruk and rice koji of As-pergillus kawachii. The odor of modified Nuruk was savory but those of commercial Nuruk and rice koji were fetid andinodorous repectively. The extracted waters of modified Nuruk commercial Nuruk and rice koji were yellow pale yellow and colorless and showed 6.15, 6.01 and 3.30 of pH respectively. Mod-ified Nuruk had 7.6${\times}$106CFU/g of yeast but commercial uruk and rice koji had no yeast. Commercial Nuruk had 1.0${\times}$102CFU/g of lactic acid bacteria but modified Nuruk and rice koji had no lactic acid bacteria. The amylolytic and proteolytic activities of modified Nuruk were much higher than those of commercial Nuruk or rice koji. Seed mash of modified Nuruk had ester aroma but that of commercial Nuruk rather offensive odor and that of rice koji neither ester aroma or offensive odor. It seemed that if the modified Nuruk is used in seed mashing the supplement of acidulant is need to lower pH. The quality of modified Nuruk was thought to be much better than that of commercial Nuruk or rice koji.

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Analysis of Mahjor Flavor Compounds in Takju Mash Brewed with a Modified Nuruk (개량누룩으로 제조한 탁주의 중요 향미성분 분석)

  • 이영숙
    • The Korean Journal of Food And Nutrition
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    • v.12 no.4
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    • pp.421-426
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    • 1999
  • The major flavor compounds of Takju mash which was brewed with a modified Nuruk made by inocu-lation and cultivation of Rhizopus japonicus T2, Aspergillus oryzae L2 and Hansenula sp. BC26 isolated from Nuruk, were analyzed, as compared with those with current fermenting agents such as commerical Nur-uk and rice koji of Aspergillus kawachii. The contents of isoamyl alcohol isobutyl alcohol and ethyl acet-ate which were known as aroma compounds in Takju were much higher in mash of modified Nuruk than in that of commercial Nuruk or ricd koji. The major organic acids were lactic fumalic and succinic acid in mash of modified and lactic and acetic acid in mash of commercial Nuruk and citric lactic and suc-cinic acid in mash of rice koji. The contents of total organic acids were 5,146mg/L, 1,706mg/L and 1, 388 mg/L in mash of commercial Nuruk rice koji and modified Nuruk respectively. The major free amino acids were glutamic acid alanine proline and histidine in mash of modified Nuruk and glutamic acid proline leucine and histidine in mash of commercial Nuruk and arginine proline and glutamic acid in mash of rice koji. The contents of total free amino acids were 14,090mg/L 12,202mg/L and 7,152 mg/L in mash of modified Nuruk commercial Nurcuk and rice koji respectively. Therefore it seemed that the Takju mash of modified Nuruk was better than that of commercial Nuruk or rice koji.

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Improvement in the Quality of Takju by a Modified Nuruk (개량누룩의 사용에 의한 탁주의 품질개선)

  • 이영숙
    • The Korean Journal of Food And Nutrition
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    • v.12 no.4
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    • pp.427-432
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    • 1999
  • Physicochemical and sensory properties of Takju which was brewed with a modified Nuruk made by inoculation and cultivation of Rhizopus japonicus T2. Aspergillus oryzae L2. and Hansenula sp. BC26 isolated from Nuruk were investigated as compared with those with current fermenting agents such as com-mercial Nuruk and rice koji of Aspergillus kawachii. The contents of total acid were 0.88% 0.47% and 0.39% in mash of commercial Nuruk rice koji and modified Nuruk respectively. The contents of alcohol were 15.5% 15.1% and 9.0% in mash of modified Nuruk rice koji and commercial Nuruk respectively. The contents of reducing sugar were 2.80% 1.24% and 0.80% in mash of commercial Nuruk modified Nuruk and rice koji respectively. The contents of amino acid were 0.55% 0.47% and 0.23% in mash of modified Nuruk commercial Nuruk and rice koji respectively. Suspended solids were sedimented much more slowly in Takju of modified Nuruk or commercial Nuruk than in that of rice koji Color was pre-ferred in the order of Takju of modified Nuruk commercial Nuruk and rice koji. Odor and taste were preferred in the order of Takju of modified Nuruk rice koji and commercial Nuruk. Therefore it seem-ed that the Takju of modified Nuruk was better than that of commercial Nuruk or rice koji in quality.

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Microbial and Enzymatic Properties Related to Brewing of Traditional Ewhaju (전통 이화주의 양조와 관련된 미생물 및 효소적 특성)

  • 김정옥;김종군
    • Korean journal of food and cookery science
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    • v.9 no.4
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    • pp.266-271
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    • 1993
  • In order to investigate the traditional value of Ewhaju (traditional wine) and to establish the brewing condition, studies on of traditional background and field inquiry were carried out. Scientific evaluation and possibility of revealation of Ewhaju were searched by the experiments of microbial and enzymatic properties of brewed Ewhaju and Nuruk by traditional method. In flora of microorganisms in Nuruk of Ewhaju, Aspergillus oryzae and Hansenula sP. were isolated, and, showed a level of 1.2$\times$$10^6$ CHU/g, respectiveln but other microorganisms were not grown in diluted cultivation test. The a-and f-amylase activity of Nuruk were 30.74 and 34.4, respectively and their activities of two amylases were 19.28 and 18.8 at first stage of brewing, 21.21 and 19.80 at 100 day after brewing, and 20.25 and 19.90 at one year aged Ewhaju, respectively. The brewed Ewhaju could be remained with high quality long period without teat treatment or addition of preservatives, also, stored Ewhaju contains remarka-bly high activity of amylases which might contribute to digestion.

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Microbiological and Enzymological Studies on Takju Brewing (탁주(濁酒) 양조(釀造)에 관(關)한 미생물학적(微生物學的) 및 효소학적(酵素學的) 연구(硏究))

  • Kim, Chan-Jo
    • Applied Biological Chemistry
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    • v.10
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    • pp.69-100
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    • 1968
  • 1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.

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