• Title/Summary/Keyword: immobilized yeast

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Application of a Compatible Xylose Isomerase in Simultaneous Bioconversion of Glucose and Xylose to Ethanol

  • Chandrakant Priya;Bisaria Virendra S.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.32-39
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    • 2000
  • Simultaneous isomerisation and fermentation (SIF) of xylose and simultaneous isomerisation and cofermentation (SICF) of glucose-xylose mixture was carried out by the yeast Saccharomyces cerevisiae in the presence of a compatible xylose isomerase. The enzyme converted xylose to xylulose and S. cerevisiae fermented xylulose, along with glucose, to ethanol at pH 5.0 and $30^{\circ}C$. This compatible xylose isomerase from Candida boidinii, having an optimum pH and temperature range of 4.5-5.0 and $30-35^{\circ}C$ respectively, was partially purified and immobilized on an inexpensive, inert and easily available support, hen egg shell. An immobilized xylose isomerase loading of 4.5 IU/(g initial xylose) was optimum for SIF of xylose as well as SICF of glucose-xylose mixture to ethanol by S. cerevisiae. The SICF of 30 g/L glucose and 70 g xylose/L gave an ethanol concentration of 22.3 g/L with yield of 0.36 g/(g sugar consumed) and xylose conversion efficiency of $42.8\%$.

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Purification of YPTP1 with Immobilized Phosphonomethylphenylalanine-Containing Peptide as an Affinity Ligand

  • Han, Jun-Pil;Kwon, Mi-Yun;Cho, Hyeong-Jin
    • BMB Reports
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    • v.31 no.2
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    • pp.135-138
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    • 1998
  • A previous study on a yeast protein tyrosine phosphatase, YPTP1, using synthetic phosphotyrosine-containing peptides with various sequences as substrates revealed that DADEpYDA exhibits high affinity ($K_m=4{\mu}M$) toward the enzyme. A modified version of this peptide, GDADEpmFDA, immobilized on a resin, was used in this study as an affinity ligand for the purification of YPTP1. Phosphonomethyl-phenylalanine (pmF) was used as a nonhydrolyzable analog of the phosphotyrosine (pY) residue, with properties similar to pY. A protected form of pmF, $Fmoc-pmF(^{t}Bu)_{2}-OH$, was chemically synthesized and introduced during solid-phase peptide sythesis. YPTP1 was onrexpressed in an E. coli strain carrying a plasmid pT7-7-ptpl. Affinity chromatography of the crude lysate afforded PTPI (39 kDa) of about 50% purity.

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Characteristics of Sulfur oxidation and the Removal of Hydrogen sulfide by Burkholdera[Pseudomonas] cepacia (Burkholderia[Pseudomonas] cepacia의 황 산화 특성 및 황화 수소 제거)

  • 정성제;이은관;전억한;윤인길;박창호
    • KSBB Journal
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    • v.16 no.5
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    • pp.466-473
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    • 2001
  • A bacterium was isolated from soils in Suwon, Korea for the purpose of H$_2$S removal using a biofilter system. The isolate was gram-negative, rod-shaped, catalase-positive, motile, and the isolated bacterium showed a positve in utilizing energy sources including citrate, mannitol, sucrose, fructors, and trehalsoe. Based on its biochemical characteristics it was identified as Burkholderia(Pseudomonas) cepacia. The growth rate of the bacterium in thiosulfate medium with yeast extract was 0.15 hr$\^$-1/ and generation time was 4.6 hr. The cell productivity was 8.05 mg/L$.$h and the isolate grew logarithmically up to 12 hr. The maximum rate of sulfur oxidation was 0.18 g-S/L$.$h. The optimum pH and temperature for the growth of the bacterium were 7.0 and 30$\^{C}$, respectively. The pH range for the growth of B. cepacia was 5.0-8.0. The oxidation rate of thiosulfate was lowered by a substrate thiosulfate when the concentration was higher than 0.12 M. both growth rate and sulfur oxidation rate of Burkholderia(Pseudomonas) cepacia was enhanced about 1.5 times with the addition of 0.2% yeast extract. The removal of hydrogen sulfide was investigated by immobilized B. cepacia with Ca-alginate. The maximum rate removal for H$_2$S was 6.25 g$.$$.$h$\^$-1/ when 12 L/h of flow rate was supplied. From this study suggest the immobilized B. cepacia could have a potential for H$_2$S removal.

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Evaluating Carriers for Immobilizing Saccharomyces cerevisiae for Ethanol Production in a Continuous Column Reactor

  • Cha, Hye-Geun;Kim, Yi-Ok;Choi, Woon Yong;Kang, Do-Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • Mycobiology
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    • v.42 no.3
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    • pp.249-255
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    • 2014
  • We evaluated a more practical and cost-effective immobilization carriers for ethanol production using the yeast Saccharomyces cerevisiae. Three candidate materials-rice hull, rice straw, and sawdust-were tested for their cell-adsorption capacity and operational durability. Derivatizations of rice hull, rice straw, and sawdust with the optimal concentration of 0.5 M of 2-(diethylamino)ethyl chloride hydrochloride (DEAE HCl) resulted in > 95% adsorption of the initial yeast cells at 2 hr for DEAE-rice hull and DEAE-sawdust and in only approximately 80% adsorption for DEAE-rice straw. In addition, DEAE-sawdust was found to be a more practical carrier for immobilizing yeast cells in terms of operational durability in shaking flask cultures with two different speeds of 60 and 150 rpm. Furthermore, the biosorption isotherms of DEAE-rice hull, -rice straw, and -sawdust for yeast cells revealed that the $Q_{max}$ of DEAE-sawdust (82.6 mg/g) was greater than that of DEAE-rice hull and DEAE-rice straw. During the 404-hr of continuous column reactor operation using yeast cells immobilized on DEAE-sawdust, no serious detachment of the yeast cells from the DEAE-sawdust was recorded. Ethanol yield of approximately 3.04 g/L was produced steadily, and glucose was completely converted to ethanol at a yield of 0.375 g-ethanol/g-glucose (73.4% of the theoretical value). Thus, sawdust is a promising practical immobilization carrier for ethanol production, with significance in the production of bioethanol as a biofuel.

Development of a Novel Yeast Strain Which Ferments Soy Sauce by Protoplast Fusion

  • Lee, Eun-Ju;Kim, Jong-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.24-30
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    • 1993
  • In order to develop a novel yeast which produces the charateristic aroma of soy sauce, a protoplast fusion between Zygosaccharomyces rouxii WFS4 and Torulopsis versatilis IAM 4993 was carried out. Auxotrophic mutants as selective markers were obtained from Zygosaccharomyces rouxii and Torulopsis versatilis by treatment of N-methyl-N -nitro-N-nitrosoguanidine. The conditions of the protoplast formation and the regeneration for fusion were examined. The protoplast fusion using polyethylene glycol 4000 led to the fusion frequency of $4~5{\times}10^{-7}\;cells/ml$. Among fusants, a fusant ST723-F31 presented the best results in terms of the aromaticity of fragrance, the growth pattern, the resistance against salt and the degree of growth according to pH. It makes easy to control the production and the balance of aroma components so that it gives a good flavor, shortens the fermentation period and, simplifies the preparation process when using a bioreactor into which fusant is immobilized.

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Immobilization of Microbial Cells and Or-ganelles by Entrapment with Urethane Prepolymers

  • Jin, Ing-Nyol
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1979.04a
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    • pp.115.4-116
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    • 1979
  • Acetone-dried cells of Arthrobacter simplex were entrapped in several preparations of hydrophilic urethane prepolymers and their steroid converting ability was examined. SeVeral solvents, such as methanol and propylone glycol, wereeffective for the conversion of hydrocortisone to prednisolone. The stability of the immobilized cells during storage and on repeated reactions was also examined. Thisconvenient entrapping method was also applicable for the immobilization of cellular organelles. yeast peroxisomes. The entrapped peroxi-somessh owed the activities of alcohol oxidase and catalase.

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Interaction of Cytochrome c and $Mn^{2+}$ -Cytochrome c Peroxidase

  • Kim, Mun-kyoung;M. Kwon;Kim, K.;Sanghwa Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.44-44
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    • 1999
  • Yeast cytochrome c peroxidase (CcP) was cloned and overexpressed in E. coli, and purified by a Ni$^{2+}$-affinity column. HoloCcP was obtained by reconstituting apoCcP with Mn$^{3+}$-protoporphyrin IX (MnPP). Electron paramagnetic resonance (EPR) spectra of spin-labeled holoCcP showed a slightly more immobilized signal than spin-labeled apoCcP.(omitted)

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Continuous Rapid Fermentation of Sardine Soy Sauce by Using Column Type Reactor Packed Immobilized Yeast Cells (고정화 효모를 충진한 column형 reactor에 의한 정어리 어간장의 속성 연속발효)

  • Kim, Seong-Joon;Shin, Dong-Bun;Ryu, Beung-Ho
    • Korean Journal of Food Science and Technology
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    • v.25 no.2
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    • pp.154-159
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    • 1993
  • This present study was carried out particulary focusing on rapid fermentation of soy sauce by using column type reactor $(30\;cm{\times}5\;cm)$ packed each immobilized cells of Pediococcus halophilus R-22, Saccharomyces rouxii R-60 and Candida etchellsii H-50. When immobilized P. halophilus R-22 by column type reactor was performed continuously fermentation, lactic acid was produced $0.62{\sim}0.64%$ during 25 days and then decreased gradually after 30 days. S. rouxii R-60 was Produced the $2.1{\sim}2.5%$ ethylalcohol constantly for 35 days and also C. etchellsii H-50 was produced $14{\sim}16\;mg/l$ 4-ethylguaiacol for 35 days and then this products were decreased gradually after fermentation of 40 days. Final Products of fish sauce contained 1,721.6 mg% total nitrogen, 1,584.1 mg% amino-nitrogen, 0.75% lactic acid, 2.7% ethylalcohol and 18.2 mg/l 4-ethylguaiacol.

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Ethanol Production by a New Method of Alginate-Immobilization (새로운 Alginate 고정화 방법에 의한 에탄올 생산)

  • Kim, Eun-Young;Kim, Seung-Wook;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.373-380
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    • 1993
  • When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

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Ethanol Production by Repeated Batch and Continuous Fermentations by Saccharomyces cerevisiae Immobilized in a Fibrous Bed Bioreactor

  • Chen, Yong;Liu, Qingguo;Zhou, Tao;Li, Bingbing;Yao, Shiwei;Li, An;Wu, Jinglan;Ying, Hanjie
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.511-517
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    • 2013
  • In this work, a fibrous bed bioreactor with high specific surface area and good adsorption efficacy for S. cerevisiae cells was used as the immobilization matrix in the production of ethanol. In batch fermentation, an optimal ethanol concentration of 91.36 g/l and productivity of 4.57 g $l^{-1}\;h^{-1}$ were obtained at an initial sugar concentration of 200 g/l. The ethanol productivity achieved by the immobilized cells was 41.93% higher than that obtained from free cells. Ethanol production in a 22-cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in packed-bed reactors, a maximum ethanol concentration of 108.14 g/l and a productivity of 14.71 g $l^{-1}\;h^{-1}$ were attained at $35^{\circ}C$, and a dilution rate of 0.136 $h^{-1}$ with 250 g/l glucose.