• Title/Summary/Keyword: High-content RNA yeast

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Selection of Yeast Mutant Strain with High RNA Content and Its High Cell-Density Fed-Batch Culture. (고함량 RNA 효모 변이주의 선별 및 고농도세포 유가배양)

  • 김재범;권미정;남희섭;김재훈;남수완
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.68-72
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    • 2002
  • To obtain a yeast mutant with high RNA content and high growth rate, Saccharomyces cerevisiae MTY62 was mutated with ethylmethane sulfonate. Among the selected mutants that were sensitive to the high concentration of KCl, M40-10 strain was finally selected due to its rapid cell growth and high RNA content in the tube and baffled-flask cultures. In the batch culture of M40-10 mutant, the maximum specific growth rate ($\mu_{max}$) of $0.38 h^{-1}$ , RNA concentration of 3210 mg-RNA/1, and RNA content of 183 mg-RNA/g-DCW were obtained, which were 23%, 15%, and 12% increased levels, respectively, compared to those of MTY62 parent strain. The intermittent fed-batch culture of M40-10 strain resulted in the maximum cell concentration of 35.6 g-DCW/1, RNA concentration of 5677 mg/1, and RNA content of 160 mg-RNA/g-DCW. Through the constant fed-batch culture, the maximum cell concentration of 46.4 g-DCW/1, RNA concentration of 6270 mg-RNA/1, and RNA content of 135 mg-RNA/g-DCW were obtained. At the 20 h culture time in the fed-batch cultures of M40-10 strain, the cell and RNA concentrations were increased by 30% and 10%, respectively, over the parent strain MTY62. In addition, it was also found that the accumulated RNA within the mutant cell was not degraded until the end of fed-batch cultivation, indicating that the M40-10 cell is a mutant with weak acidic RNase activity.y.

Fed-batch Production of High-Content RNA Yeast by Using Industrial Medium

  • Kim, Jae-Beom;Nam, Su-Wan
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.295-298
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    • 2000
  • In order to maximize the RNA accumulation and biomass production in Saccharomyces cerevisiae MTY62, a high-content RNA yeast strain, fed-bach cultures were performed with optimized industrial medium including molasses and corn steep liquor. Among the feeding modes examined, the constant feeding mode resulted in the cell concentration of 35.7 g-DCW/L and the RNA concentration of 5434 ${\mu}g-RNA/mL$, which were about 2-fold increased levels, compared to the results of bach culture. However, the RNA content (153 mg-RNA/g-DCW) in the fed-batch cultures was lower than that in the batch culture (171 mg-RNA/g-DCW).

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Fed-Batch Fermentation of High-Content RNA Yeast by Using Molasses Medium. (당밀 배지를 이용한 고함량 RNA효모의 유가배양)

  • 김재범;권미정;남희섭;김재훈;남수완
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.234-239
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    • 2001
  • In order to maximize the RNA accumulation and biomass production is Saccharomyces cerevisiae MTY62, a high-content RNA yeast strain, batch and fed-batch cultures were performed. Among the feeding modes of fed-batch cultures examined, the intermittent feeding mode R\`(IFB-lV), in which 50 ml of 40% molasses and 20% com steep liquor (CSL) solution was intermittently fed for 5 times, resulted in the cell concentration of 33.8 g- dry cell weight/1 and the RNA concentration of 5221 mg-/l, and RNA content of 153 mg-RNA/g-dry cell weight. The constant fed-batch with feeding mode III (CFB-III), in which the feeding rate of 40% molasses and 20% CSL solution was stepwisely decreased from 48 mph (9-13 h), to 24 mph (13-21 h), and to 18 ml/h (21∼ 48 h), gave the highest cell concentration of 42.7 g-dry ceil weigh71 and R7IA concentration of 5536 mg-RNA/1, which were about 2.4-fold and 1.9-fold increased levels, respectively, compared to the results of batch culture. However, the RNA con- tent of 130 mg-RNA/g-dry cell weight of the fed-batch was lower than that of the batch culture (171 mg-RNA/g-dry cell weight) and other fed-batch cultures. When the specific growth rates in the fed-batch cultures were increased, the RNA contents increased. This result indicates that the RNA content is adversely proportional to the cell concen- tration. However, at the same specific growth rate, the RNA content was maintained at higher level in the intermit- tent fed-batch than in the constant fed-batch culture.

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RNA Production from the Hydrolyzate of Sliced and Dried Sweet Potatoes. (절간고구마를 이용한 RNA 생산에 관한 연구)

  • 이종임;송재철;조원대;양한철
    • Microbiology and Biotechnology Letters
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    • v.8 no.1
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    • pp.9-18
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    • 1980
  • During an extensive screening tests of yeasts for their RNA formation, it was found that Cryptococcus laurentii had especially high RNA content and high dry cell weight, when hydrolyzate of sliced and dried sweet potatoes was used as a carbon source. Growth conditions of this strain were examined, and the most desirable results were obtained at 48 hours of cultivation on a reciprocal shaker at 3$0^{\circ}C$ with initial pH 6.0. Under the above conditions, the RNA content and yield of dry cells were investigated using various media compositions. Ammonium sulfate 0.40%, peptone 0.6 %, and yeast extract 0.4% were appeared to be favorable as a nitrogen sources. The optimum concentrations of K $H_2$P $O_4$, M $n^{++}$, C $O^{++}$ were 0.05 %, 0.1 %, and 0.001 %, respectively. Ca-pantothenate, 400$\mu\textrm{g}$/$m\ell$, showed relatively favorable effects as a growth factor. The maximum RNA content obtained in this study was 16.8 % of the total dry cell weight.t.t.

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Preparation of Flavor-enhancing Yeast Extract Using a Saccharomyces cerevisiae Strain with High RNA Content (고핵산 함유 Saccharomyces cerevisiae 균주를 이용한 정미성 효모 추출물의 제조)

  • Kim, Jae-Sik;Kim, Jin-Wook;Shim, Won;Kim, Jung-Wan;Park, Kwan-Hwa;Pek, Un-Hua
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.475-481
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    • 1999
  • Yeast extracts were prepared using either autolysis or enzymatic digestion methods for industrial application of the Saccharomyces cerevisiae B24 strain developed previously to have high RNA content. Extraction ratio of yeast extract from yeast cell reached 65% when autolysis of yeast slurry having 10% solid content was induced at $50^{\circ}C$ and pH 5.0 by agitating with 100 rpm. However, neither 5'-IMP nor 5'-GMP was detected from the autolyzate. In another attempt to prepare a yeast extract S. cerevisiae B24 culture was treated at $90^{\circ}C$ and then treated by various enzymes including ${\beta}-1,3-glucanase$, phosphodiesterase (nuclease P1), adenylic deaminase, and a protease. The yeast extract prepared by the enzymatic digestion method contained 3.2g of 5'-IMP and 5'-GMP/100g dry yeast extract.

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Effect of Ca^{++} and Cu^{++} Removal from Molasses on Yest cell Growth and RNA Accumulation. (당밀로부터 Ca^{++} 및 Cu^{++} 이온 제거가 효모 생육 및 RNA축적에 미치는 영향)

  • 김재범;허선연;김중균;남희섭;남수완
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.211-215
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    • 2003
  • When Saccharomyces cerevisiae MTY62, a high-RNA content yeast, was cultivated by fed-batch mode feeding molasses and com steep liquor, the cell density less than 45g-DCW/L and the RNA content less than 140mg/g-cell were obtained, indicating that unknown compounds inhibiting the cell growth and RNA accumulation are contained in the molasses. Therefore, in order to obtain higher levels of cell density and RNA content, $Ca^{++}$, $Cu^{++}$and $K^{+}$ ions in molasses were removed by pretreatments of molasses with various agents such as IonClear BigBead, $Na_2$$HPO_4$, $H_2$$SO_4$, citric acid, $K_2$$HPO_4$, and EDT A. Among them, IonClear BigBead, $Na_2$$HPO_4$, and EDTA gave the highest $Ca^{++}$ removal efficiency of about 60-90%. In the batch culture with pretreated molasses, the cell concentration of 18.6g-DCW/I and RNA concentration of 3127 mg/I, maximum specific growth rate of 0.459$h^{-1}$ , and specific consumption rate of reducing sugar of 1.28g-sugar/g-cell-h were obtained, which are about 10%, 17%,47%, and 36% higher levels, respectively, over the batch culture with untreated molasses.

Identification of Lactic Acid Bacteria Involved in Traditional Korean Rice Wine Fermentation

  • Seo, Dong-Ho;Jung, Jong-Hyun;Kim, Hyun-You;Kim, Young-Rok;Ha, Suk-Jin;Kim, Young-Cheul;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • v.16 no.6
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    • pp.994-998
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    • 2007
  • Changes in microflora, pH, reducing sugar content, lactic acid content, and ethanol content during Korean rice wine fermentation were investigated. Typical quality characteristics of Korean rice wine fermentation including pH, reducing sugar content, lactic acid content, and ethanol content were evaluated. While a fungus was not detected in our Korean rice wine mash, yeast was found to be present at fairly high quantities (1.44-4.76\;{$\times}\;10^8\;CFU/mL$) throughout the fermentation period. It is assumed that lactic acid bacteria (LAB) had effects on the variations of fragrance and flavor for traditional Korean rice wine. The main LAB during the Korean rice wine fermentation was determined and identified as a Gram-positive, straight rod-shaped cell. Genotypic identification of the isolated strain by amplification of its 16S rRNA sequence revealed that the isolated strain was most closely related to Lactobacillus plantarum (99%) strains without any other comparable Lactobacillus strains. Therefore, we designated the major LAB identified from traditional Korean rice wine fermentation as L. plantarum RW.

Development of Saccharomyces cerevisiae Strains with High RNA Content (리보핵산을 다량으로 함유하는 Saccharomyces cerevisiae 균주의 개발)

  • Kim, Jae-Sik;Kim, Jin-Wook;Shim, Won;Min, Byoung-Cheol;Kim, Jung-Wan;Park, Kwan-Hwa;Pek, Un-Hua
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.465-474
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    • 1999
  • RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

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Identification and Characterization of an Oil-degrading Yeast, Yarrowia lipolytica 180

  • Kim, Tae-Hyun;Lee, Jung-Hyun;Oh, Young-Sook;Bae, Kyung-Sook;Kim, Sang-Jin
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.128-135
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    • 1999
  • Among oil-degrading microorganisms isolated from oil-polluted industrial areas, one yeast strain showed high degradation activity of aliphatic hydrocarbons. From the analyses of 18S rRNA sequences, fatty acid, coenzyme Q system, G+C content of DNA, and biochemical characteristics, the strain was identified as Yarrowia lipolytica 180. Y. lipolytica 180 degraded 94% of aliphatic hydrocarbons in minimal salts medium containing 0.2% (v/v) of Arabian light crude oil within 3 days at 25$^{\circ}C$. Optimal growth conditions for temperature, pH, NaCl concentration, and crude oil concentration were 30$^{\circ}C$, pH 5-7, 1%, and 2% (v/v), respectively. Y. lipolytica 180 reduced surface tension when cultured on hydrocarbon substrates (1%, v/v), and the measured values of the surface tension were in the range of 51 to 57 dynes/cm. Both the cell free culture broth and cell debris of Y. lipolytica 180 were capable of emulsifying 2% (v/v) crude oil by itself. They were also capable of degrading crude oil (2%). The strain showed a cell surface hydrophobicity higher than 90%, which did not require hydrocarbon substrates for its induction. These results suggest that Y. lipolytica has high oil-degrading activity through its high emulsifying activity and cell hydrophobicity, and further indicate that the cell surface is responsible for the metabolism of aliphatic hydrocarbons.

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Intestinal microbial composition changes induced by Lactobacillus plantarum GBL 16, 17 fermented feed and intestinal immune homeostasis regulation in pigs

  • Da Yoon, Yu;Sang-Hyon, Oh;In Sung, Kim;Gwang Il, Kim;Jeong A, Kim;Yang Soo, Moon;Jae Cheol, Jang;Sang Suk, Lee;Jong Hyun, Jung;Jun, Park;Kwang Keun, Cho
    • Journal of Animal Science and Technology
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    • v.64 no.6
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    • pp.1184-1198
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    • 2022
  • In this study, Rubus coreanus (R. coreanus) byproducts with high polyphenol content were fermented with R. coreanus-derived lactic acid bacteria (Lactobacillus plantarum GBL 16 and 17). Then the effect of R. coreanus-derived lactic acid bacteria fermented feed (RC-LAB fermented feed) with probiotics (Bacillus subtills, Aspergillus oryzae, Yeast) as a feed additive for pigs on the composition of intestinal microbes and the regulation of intestinal immune homeostasis was investigated. Seventy-two finishing Berkshire pigs were randomly allotted to four different treatment groups and 18 replicates. RC-LAB fermented feed with probiotics increased the genera Lactobacillus, Streptococcus, Mitsuokella, Prevotella, Bacteroides spp., Roseburia spp., and Faecalibacterium prausnitzii, which are beneficial bacteria of the digestive tract of pigs. Also, RC-LAB fermented feed with probiotics decreased the genera Clostridium, Terrisporobacter, Romboutsia, Kandleria, Megasphaera and Escherichia, which are harmful bacteria. In particular, the relative abundance of the genera Lactobacillus and Streptococcus increased by an average of 8.51% and 4.68% in the treatment groups and the classes Clostridia and genera Escherichia decreased by an average of 27.05% and 2.85% in the treatment groups. In mesenteric lymph nodes (MLN) and spleens, the mRNA expression of transcription factors and cytokines in Th1 and Treg cells increased and the mRNA expression of Th2 and Th17 transcription factors and cytokines decreased, indicating a regulatory effect on intestinal immune homeostasis. RC-LAB fermented feed regulates gut immune homeostasis by influencing the composition of beneficial and detrimental microorganisms in the gut and regulating the balance of Th1/Th2 and Th17/Treg cells.