• Title, Summary, Keyword: recombinant yeast

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Repair of Iron Deficiency in Rats by the Intake of Recombinant Yeast Producing Human H-ferritin (훼리틴 생산 재조합 효모의 철분 결핍성 빈혈 개선 효과)

  • Hwang Eun-Hee
    • Journal of the East Asian Society of Dietary Life
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    • v.16 no.1
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    • pp.93-98
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    • 2006
  • This study examined whether or not the iron that is accumulated in the recombinant microbes that produce ferritin is bioavailable to rats with iron deficiency. Rats induced with iron deficiency were treated with iron preparations of $Fe(NH_4)_2(SO_4)_2$, horse spleen ferritin, control yeast, and ferritin-producing recombinant yeast for 14 days. The bioavailability of iron was examined by measuring hemoglobin concentration, hematocrit value, and tissue iron stores. Differences between dietary groups were determined by one-way ANOVA, at the level of significance p<0.05. Based on hemoglobin concentration and hematocrit value, iron in $Fe(NH_4)_2(SO_4)_2$, horse spleen ferritin, and ferritin-producing yeast were bioavailable in rats and cured iron deficiency. The efficacy of ferritin and ferritin-producing yeast was confirmed in establishing tissue iron stores after the induction of iron deficiency. The iron sources of ferritin and the ferritin-producing yeast seemed to be as effective for the recovery from iron deficiency as the iron compounds of ferric citrate and ferrous ammonium sulfate. The results suggest that the iron stored in ferritin of the recombinant yeast is bioavailable, and that the recombinant yeast may contribute widely as a source of iron to resolve the global problem of iron deficiency.

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Applications of Yeast Flocculation in Biotechnological Processes

  • Domingues, Lucilia;Vicente, Antonio A.;Lima, Nelson;Teixeira, Jose A.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.288-305
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    • 2000
  • A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects-the basics of yeast flocculation, the development of "new" flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous $\beta$-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculating bioreactors and discussing potential new uses of these systems.e systems.

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Screening of Sterol Biosynthesis Inhibitors from Natural Products Using Recombinant Yeast Carrying Human Lanosterol Synthase

  • Sung, Chung-Ki;Kim, Eun-A;Chu, Yun-Ho;Shibuya, Masaaki;Ebizuka, Yutaka
    • Natural Product Sciences
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    • v.9 no.4
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    • pp.299-303
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    • 2003
  • For the screening of inhibitors of sterol biosynthesis from natural products, a simple and rapid assay method was developed using recombinant yeast carrying human lanosterol synthase, main target of this assay method. Sterol biosynthesis inhibition activity was monitored only by the inhibition of growth of the recombinant yeast. By changing the substrate, this assay method can figure out which step is inhibited in the sterol biosynthesis by the test material. With this assay method total 102 plant samples were screened for their inhibitory activity of sterol biosynthesis. Among plant water extracts screened, 11 plant samples showed inhibitory activity on sterol biosynthesis in ergosterol (-) medium. For selection of the specific inhibitory materials, 11 plant samples were reassayed in ergosterol (+) medium. After all 5 plant samples, Abutilon avicennae Gaertn. (stem), Alnus japonica Steud. (stem), Amaranthus mangostanus L. (aerial part), Philadelphus schrenckii Pupr. (leaf) and Pimpinelia brachycarpa Nakai (aerial part), showed specific inhibitory activity.

Hormonal Effects of Several Chemicals in Recombinant Yeast, MCF-7 Cells and Uterotrophic Assays in Mice

  • Park, Jin-Sung;Lee, Beom-Jun;Kang, Kyung-Sun;Tai, Joo-Ho;Cho, Jae-Jin;Cho, Myung-Haing;Inoue, Tohru;Lee, Yong-Soon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.293-299
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    • 2000
  • Many methods have been developed for screening chemicals with hormonal activity. Using recombinant yeasts expressing either human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127 (YER)] or androgen receptor [S. cerevisiae AR + 8320 (YAR)], we evaluated the hormonal activities of several chemicals by induction of ${\beta}-galactosidase$ activity. The chemicals were $17{\beta}-estradiol$ (E2), testosterone (T), ${\rho}-nonylphenol$ (NP), bisphenol A (BPA), genistein (GEN), 2-bromopropane (2-BP), dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and butylparaben (BP). To assess the estrogenicity of NP, the result of the in vitro recombinant yeast assay was compared with an E-screen assay using MCF-7 human breast cancer cells and an uterotrophid assay using ovariectomized mice. In the YER yeast cells, E2, NP, BPA, GEN, and BP exhibited estrogenicity in a doseresponse manner, while TCDD did not. All the chemicals tested, except T, did not show androgenicity in the YAR yeast cell. The sensitivity of the yeast (YER) assay system to the estrogenic effect of NP was similar to that of the E-screen assay. NP was also estrogenic in the uterotrophic assay. However, in terms of convenience and costs, the yeast assay was superior to the E-screen assay or uterotrophic assay. These results suggest that the recombinant yeast assay can be used as a rapid tool for detecting chemicals with hormonal activities.

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Recombinant Human L-ferritin from Saccharomyces cerevisiae: Molecular Characterization and Synthesis of Iron Oxide Nanoparticles (효모에서 생산한 재조합 human L-ferritin의 생화학적 특성 및 나노입자의 철산화물 합성)

  • Kim, Kyung-Suk
    • KSBB Journal
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    • v.26 no.2
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    • pp.119-125
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    • 2011
  • In the synthesis of nanoparticles, much attention has been paid to regulating the particle size. There has been a possible evident that using the central cavity (core) of the protein ferritin has a greatly significant influence on it because the core can generate the nanometer-sized mineral particles of variable metal ions. In this report, recombinant human L-ferritins produced from Saccharomyces cerevisiae were purified and their molecular properties were characterized. The cDNA for human ferritin L chain was also expressed in another host such as Escherichia coli, and the properties of recombinant L-ferritins were compared. From isoelectric focusing experiment, the L-ferritin from the recombinant yeast showed no indication of N-glycosylation. Some post-translational modifications other than N-glycosylation were speculated in the L-ferritins from yeast. A difference was made in the L-ferritins in their iron uptake rates and the initial rate of the L-ferritin from yeast was slightly increased. The reconstitution yield and size distribution of the core minerals were analyzed in the L-ferritins by transmission electron microscopy. The L-ferritin from yeast with higher reconstitution yield (54.5%) showed slightly larger sizes (mean 6.92 nm) with narrower size distribution than the L-ferritin from E. coli. It is, in conclusion, speculated that L-ferritin from yeast is relatively superior to the other, in view of the size of nanoparticle and its relative homogeneity.

Development of Cellobiose-utilizing Recombinant Yeast for Ethanol Production from Cellulose Hydrolyzate

  • Pack, Seung-Pil;Cho, Kwang-Myung;Kang, Hyen-Sam;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.441-448
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    • 1998
  • A cellobiose-utilizing recombinant yeast having $\beta$-glucosidase activity was developed for ethanol production from a mixture of glucose and cellobiose. Using $\delta$-sequences of Tyl transposon of yeast as target sites for homologous recombination, a heterologous gene of $\beta$-glucosidase was integrated into the chromosome of Saccharomyces cerevisiae. The $\delta$-integrated recombinant yeast, Saccharomyces cerevisiae L2612 (Pb-BGL), showed perfect mitotic stability even in nonselective media and showed ca. 1.5 fold higher $\beta$-glucosidase activity than the recombinant yeast harboring the $2\mu$-based plasmid vector system. A mathematical model was developed to describe the $\beta$-glucosidase formation and ethanol production from the Saccharomyces cerevisiae L2612 ($p\delta-BGL$). The model newly described that the heterologous $\beta$-glucosidase production mediated by ADH1 promoter is regulated by glucose and repressed by ethanol.

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A Study of Immunogenicity and Reactogenicity of Hepatitis B Vaccine Made by Recombinant DNA Techniques in Yeast (효모재조합 DNA B형 간염백신의 면역효과에 관한 연구)

  • Min, Chang-Hong;Kim, Kyo-Myung;Lee, Kyu-Man
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.2
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    • pp.243-249
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    • 1986
  • A study of the immunogenicity and reactogenicity of two doses of lot H(10, 20 mcg), two doses of lot L (20, 40 mcg) of the Smith Kline-RIT recombinant DNA yeast-derived hepatitis B vaccine and a 20-mcg dose of the Merck Sharp and Dohme plasma-derived hepatitis B vaccine was conducted in young adults under randomized, double-blind conditions. Immunization was carried out according to a 0-, 1-, and 6-month vaccination schedule. Results indicated that the yeast-derived hepatitis B vaccine was well tolerated and immunogenic. Reactogenicity to both yeast- and plasma-derived vaccines was mild in severity and low in incidence with no significant differences appearing between the study groups. One month after the third dose, the yeast-derived vaccines induced a high degree of soroconversion ranging between 95.0% and 100%. The response was not lot or dose-dependent. The administration of the plasma-derived vaccine resulted in anti-HBs geometric mean titres statistically signifirantly higher than those elicited by the different yeast-derived hepatitis B vaccines one month after the third dose of vaccine but the difference was not large enough to be of great clinical significance.

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Production of ${\gamma}$-Glutamylcysteine by Immobilized Mixed Microbial System of Recombinant E. coli and Yeast (재조합 대장균과 효모의 고정화 혼합세포계에 의한 ${\gamma}$-Glutamylcysteine 생산)

  • 김원근;구윤모
    • KSBB Journal
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    • v.10 no.3
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    • pp.249-256
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    • 1995
  • ${\gamma}$-Glutamylcysteine production by the immobilized microbial system of recombinant Escherichia coli and yeast was investigated. ${\gamma}$-Glutamylcysteine was synthesized from L-glutamic acid and L-cysteine in the presence of ATP by the reaction catalyzed by ${\gamma}$-glutamylcysteine synthetase. An immobilized microbial cell system was developed for the efficient ${\gamma}$-glutamylcysteine production. Recombinant Escherichia coli and yeast were immobilized by alginate. Production of ${\gamma}$-glutamylcysteine was better with the recombinant Escherichia coli for both the synthesis of ${\gamma}$-glutamylcysteine and the ATP regeneration than the mixed system of recombinant Escherichia coli and yeast. The proper radio of recombinant Escherichia coli to yeast was experimentary observed to be 1:4 in the mixed system. Although the immobi1ized system had the slower reaction rate, its reaction stability was increased by 10%.

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Fermentation Strategies for Recombinant Protein Expression in the Methylotrophic Yeast Pichia pastoris

  • Zhang, Senhui;Inan, Mehmet;Meagher, Michael M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.275-287
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    • 2000
  • Fermentation strategies for recombinant protein production in Pichia pastoris have been investigated and are reviewed here. Characteristics of the expression system, such as phenotypes and carbon utilization, are summarized. Recently reported results such as growth model establishment, app58lication of a methanol sensor, optimization of substrate feeding strategy, DOstat controller design, mixed feed technology, and perfusion and continuous culture are discussed in detail.

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Iron-fortified recombinant Saccharomyces cerevisiae producing Sus scrofa ferritin heavy-chain recovers iron deficiency in mice

  • Lim, Hwan;Kim, Jong-Taek;Kim, Myoung-Dong;Rhee, Ki-Jong;Jung, Bae Dong
    • Korean Journal of Veterinary Research
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    • v.52 no.4
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    • pp.263-268
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    • 2012
  • In this study, we produced iron-fortified yeast (Saccharomyces cerevisiae) producing Sus scrofa ferritin heavy-chain to provide iron supplementation in anemic piglets. We determined whether iron-ferritin accumulated in recombinant yeasts could improve iron deficiency in mice. C57BL/6 male mice exposed to Fe-deficient diet for 2 weeks were given a single dose of ferrous ammonium sulfate (FAS), ferritin-producing recombinant yeast (APO), or APO reacted with iron ($Fe^{2+}$) (FER). The bioavailability of recombinant yeasts was examined by measuring body weight gain, hemoglobin concentration and hematocrit value 1 week later. In addition, ferritin protein levels were evaluated by western blot analysis and iron stores in tissues were measured by inductively coupled plasma spectrometer. We found that anemic mice treated with FER exhibited increased levels of ferritin heavy-chain in spleen and liver. Consistently, this treatment restored the iron concentration in these tissues. In addition, this treatment significantly increased hemoglobin value and the hematocrit ratio. Furthermore, FER treatment significantly enhanced body weight gain. These results suggest that the iron-fortified recombinant yeast strain is bioavailable.