• Title/Summary/Keyword: Rhizopus oryzae

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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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In vitro Antifungal Activity of 4-Hydroxyderricin and Acetylshikonin against Ascosphaera apis

  • Park, Sangchul;Shin, Yu-Kyong;Cho, MyoungLae;Kwon, Hyun Sook;Kwon, Yun Ju;Kim, Ki-Young
    • Journal of Apiculture
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    • v.34 no.2
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    • pp.125-129
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    • 2019
  • Honey bees are important pollinators in agriculture, but are threatened by the pathogen Ascosphaera apis, which causes chalkbrood. Despite attempts to control this fungus using synthetic fungicides, none of them have been proven to be completely effective. Among 640 natural compounds that we tested, 4-hydroxyderricin (MIC=3.125, 6.25 mg/L after 24 h and 48 h growth, respectively) exhibited the strongest anti- Ascosphaera apis activity, followed by acetylshikonin (MIC=12.5 mg/L for 24 h and 48 h growth). 4-Hydroxyderricin showed selective growth inhibition of Ascosphaera apis and Rhizopus oryzae among tested fungus strains. Treatment 4-hydroxyderricin with miconazole revealed a synergistic effect (FICI=0.65±0.13 at 48 h incubation). These findings suggest that 4-hydroxyderricin, which has antifungal activity against Ascosphaera apis but few other fungal species, can effectively control infectious fungal diseases. Combined treatment of bees with 4-hydroxyderricin and miconazole could reduce cytotoxicity and improve the cost effectiveness of treatment.

Chemical Characteristics in Fermented Cordycepin-enriched Cordyceps militaris (발효 동충하초의 이화학적 특성)

  • Ahn, Hee-Young;Park, Kyu-Rim;Kim, Yu-Ra;Cha, Jae-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1032-1040
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    • 2013
  • The comparative effects of fibrinolytic, antioxidative activity and electrophoretical protein patterns with Cordyceps militaris powder (CM) and cordycepin-enriched Cordyceps militaris JLM0636 powder (CCM) and fermented cordycepin-enriched Cordyceps militaris JLM0636 powder by several microscopic organisms were investigated. In addition, nutritional materials such as mineral, protein, and fatty acids were also measured. The protein concentration was higher in CCM than that in CM. The protein concentration in fermented CCM was the highest in CCM treated by Aspergillus kawachii among the various samples. When total protein patterns of CM, CCM, and both fermented CCMs were analyzed by native- and SDS-PAGE, there were slightly varietal differences in electrophoretical protein patterns. Major minerals were K, Ca, Mg, and Zn. Major fatty acids were palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Fibrinolytic activity was the highest in the fermented CCM by Bs treatment among the various samples. The ${\alpha},{\alpha}^{\prime}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radical scavenging activity was slightly stronger in the CCM treated with Aspergillus kawachii among the various samples; however, these samples all exhibited relatively low levels of activity compared with the butylated hydroxytoluene (BHT). These results may provide the basic data necessary to understand the biological activities and chemical characteristics of Cordyceps militaris JLM0636 powder fermented by several microscopic organisms to develop functional foods.

Production of Gamma-Linolenic Acid in Pichia pastoris by Expression of a Delta-6 Desaturase Gene from Cunninghamella echinulata

  • Wan, Xia;Zhang, Yinbo;Wang, Ping;Huang, Fenghong;Chen, Hong;Jiang, Mulan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1098-1102
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    • 2009
  • Gamma-linolenic acid (GLA, C18:3 ${\Delta}^{6,9,12}$) is synthesized by a delta-6 fatty acid desaturase using linoleic acid (LA, C18:2 ${\Delta}^{9,12}$) as a substrate. To enable the production of GLA in the conventional yeast Pichia pastoris, we have isolated a cDNA encoding the delta-6 fatty acid desaturase from Cunninghamella echinulata MIAN6 and confirmed its function by heterogeneous expression in P. pastoris. Sequence analysis indicated that this cDNA sequence has an open reading frame of 1,404 bp, which encodes a 52 kDa peptide of 468 amino acids. This sequence has 64% identity to the previously reported delta-6 fatty acid desaturase from Rhizopus oryzae. The polypeptide has a cytochrome b5 domain at the N-terminus including the HPGG motif in the heme-binding region, as reported for other delta-6 fatty acid desaturases. In addition, this enzyme differs from other desaturases by the presence of three possible N-linked glycosylation sites. Analysis of the fatty acid composition demonstrated the accumulation of GLA to the level of 3.1% of the total fatty acids. Notably, the amounts of ginkgolic acid (C17:1) and palmitic acid (C16:0) were increased from 1.3% to 29.6% and from 15% to 33%, respectively. These results reveal that the modification of the fatty acid biosynthetic pathway by genetic manipulation in order to produce specific polyunsaturated fatty acids in P. pastoris is a promising technique.

Bioproduction of trans-10,cis-12-Conjugated Linoleic Acid by a Highly Soluble and Conveniently Extracted Linoleic Acid Isomerase and an Extracellularly Expressed Lipase from Recombinant Escherichia coli Strains

  • Huang, Mengnan;Lu, Xinyao;Zong, Hong;Zhuge, Bin;Shen, Wei
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.739-747
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    • 2018
  • The low solubility and high-cost recovery of Propionibacterium acnes polyunsaturated fatty acid isomerase (PAI) are key problems in the bioproduction of high value-added conjugated linoleic acid (CLA). To improve the solubility of recombinant PAI, six chaperone proteins were coexpressed with PAI. Introduction of GroELS proteins dramatically improved the PAI solubility from 29% to 97%, with increased activity by 57.8%. Combined expression of DnaKJ-GrpE and GroELS proteins increased the activity by 11.9%. In contrast, coexpression of DnaKJ-GrpE proteins significantly reduced the activity by 57.4%. Plasmids pTf16 harboring the tig gene and pG-Tf2 containing the tig and groEL-groES genes had no visible impact on PAI expression. The lytic protein E was then introduced into the recombinant Escherichia coli to develop a cell autolysis system. A 35% activity of total intracellular PAI was released from the cytoplasm by suspending the lysed cells in distilled water. The PAI recovery was further improved to 81% by optimizing the release conditions. The lipase from Rhizopus oryzae was also expressed in E. coli, with an extracellular activity of 110.9 U/ml. By using the free PAI and lipase as catalysts, a joint system was established for producing CLA from sunflower oil. Under the optimized conditions, the maximum titer of t-10,c-12-CLA reached 9.4 g/l. This work provides an effective and low-cost strategy to improve the solubility and recovery of the recombinant intracellular PAI for further large-scale production of CLA.

Inhibition of Bacillus cereus in Doenjang Fermented with Multiple Starters Showing Inhibitory Activity against Pathogens (유해균 억제능을 지닌 복합종균을 접종하여 제조한 된장에서 Bacillus cereus 억제)

  • Lee, Jae Yong;Shim, Jae Min;Lee, Kang Wook;Cho, Kye Man;Kim, Gyoung Min;Shin, Jung-Hye;Kim, Jong-Sang;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.254-260
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    • 2016
  • Doenjang samples were prepared by inoculation of multiple starters consisting of two Bacillus spp., one yeast, and one fungus. Doenjang A was fermented with Bacillus amyloliquefaciens EMD17, B. amyloliquefaciens MJ1-4, Pichia farinosa SY80, and Rhizopus oryzae. Doenjang B and C were fermented with the same yeast and fungus but different Bacillus strains; namely, B. amyloliquefaciens EMD17 and B. subtilis CH3-5 for doenjang B, and B. amyloliquefaciens MJ1-4 and B. subtilis CH3-5 for doenjang C. Doenjang D was fermented with microorganisms present in rice straw (control). The doenjang samples were spiked with B. cereus ATCC14579 at two different levels, 104 CFU/g doenjang (I) and 107 CFU/g doenjang (II). All eight doenjang samples were fermented for 70 days at 25℃. Growth of B. cereus was inhibited in doenjang A, B, and C, with the bacterial cell count after 70 days being less than the initial 104 CFU/g added, whereas B. cereus was not inhibited in doenjang D. Doenjang B showed the strongest inhibitory activity against B. cereus, with a cell count of less than 103 CFU/g after 42 days, even when B. cereus was initially added at 107 CFU/g. Some properties of the doenjang samples, such as pH, TA, and amino-type nitrogen content, were similar to those of doenjang fermented with starters only. The results indicate that carefully selected starters can effectively prevent the growth of B. cereus during doenjang fermentation.

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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