• Applied Biological Chemistry
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• v.39 no.1
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• pp.1-8
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• 1996

A one-stage, continuous-flow bioreactor with both immobilized and suspended cells was used to investigate the roles of lipid supplements in ethanol production by Saccharomyces cerevisiae. The reactor performance and the level of alcohol dehydrogenase(ADH) activities of the suspended cells, grown under various conditions, were measured. When ergosterol and/or oleic acid were added with surfactants to the yeast culture grown under non-aerated conditions, remarkable increases in ethanol production and cell growth was achieved, but specific ADH activities were not affected. Especially, no difference of specific ADH activities of the suspended cells grown under aerated and non-aerated condition was observed. The addition of the surfactant as a supplement also resulted in significant increases in ethanol production, cell growth, and specific ADH activity. When ergosterol and oleic acid were added to the yeast culture exposed to higher ethanol concentration($>40\;g/{\ell}$) level, ethanol production, cell growth, and specific ADH activity were increased, but the addition of surfactant was as effective as at lower ethanol concentration level. The results indicated that lipid supplements were more effective at higher ethanol concentration level than at lower ethanol concentration level during ethanol production. ADH isozyme patterns of the yeast cultures grown under various conditions on starch gel electrophoresis showed only one major band, probably ADH I. The migrating distance of the major isozyme, however, varied slightly according to the culture conditions of the cells. No apparent correlation was found between specific ADH activity and amount of ethanol produced. Cell mass was more important factor for ethanol production than specific ADH activity of the cells.

• Bulletin of the Korean Chemical Society
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• v.32 no.12
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• pp.4337-4340
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• 2011

The vitamin D receptor binding peptide, VDRBP, was overexpressed as a fused form with the ubiquitin molecule in Rosetta(DE3)pLysS, a protein production strain of Escherichia coli harboring an induction controller plasmid. The fusion protein was bound to the immobilized metal ions, and the denaturation and renaturation of the fusion protein were performed as a part of the purification procedure. After the elution of the fusion protein, the peptide hormone was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The purity of the resulting peptide fragment was checked by MALDI-TOF mass and NMR spectroscopy. The final yields of the target peptide were around 5 and 2 mg per liter of LB and minimal media, respectively. The recombinant expression and purification of this peptide will enable structural and functional studies using multidimensional NMR spectroscopy and X-ray crystallography.

• Bulletin of the Korean Chemical Society
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• v.28 no.9
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• pp.1549-1552
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• 2007

The cytoplasmic domain of syndecan-4, a type I transmembrane heparan sulfate proteoglycan, was overexpressed as a fused form with the ubiquitin molecule in Escherichia coli, and the fusion protein was purified using immobilized metal affinity chromatography (IMAC). The cytoplasmic domain was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The integrity of the resulting peptide fragment was checked by MALDI-TOF and NMR spectroscopy. The yield of the peptide was 3.0-1.5 mg per liter in LB or minimal medium, respectively. The recombinant expression and purification of this domain will enable us its structural and functional studies using multidimensional NMR spectroscopy.

• Journal of the Korean Ceramic Society
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• v.39 no.9
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• pp.857-862
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• 2002

Porous ceramic supports with immobilized microorganisms for the water purifier were synthesized by firing green compacts of mixed powder comprising of fly ash, bentonite and an additive of yeast powder at 800∼1,000$^{\circ}C$ for 1h and the pore and mechanical properties of specimens were investigated. The compressive strength was increased in FB (Fly Ash + Bentonite) specimens while pore properties was decreased with increasing the bentonite content and sintering temperature. The compressive strength, bulk density, apparent density, porosity, mean pore size, pore volume and specific surface area of FB specimens at 800∼1,000$^{\circ}C$ were 89.6∼128.9 kgf/$cm^2$, 1.25∼1.43, 1.61∼1.78, 27.2∼62.2%, 7.9∼25.6 ${\mu}m$, 8.9∼$22.2{\times}10^{-5}\;cm^3/g$ and 35.2∼134.3 $m^2/g$, respectively. The pore properties of FBY (FB+yeast powder) specimens were superior to that of FB specimens, however compressive strength was decreased with increasing yeast powder content. The overall properties of 9F1B1Y (9F1B+10% of yeast powder) specimens at 900$^{\circ}C$ for 1 h were 98.7 kgf/$cm^2$, 1.20, 1.67, 68.1%, 48.9 ${\mu}m$, $29.5{\times}10^{-5}\;cm^3/g$ and 152.2 $m^2/g$, respectively. In this study, it was revealed that 9F1B1Y specimen demonstrated better S. saprophyticus adherence properties n their surface pores. Consequently, the microorganisms immobilized on porous ceramic supports showed better water purifying performance with many pores and adequate strength.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.142-149
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• 1991

Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.10
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• pp.1417-1422
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• 2012

Alkyl glucosides were synthesized using the transglycosylation reaction of Celluclast, the cellulase from Trichoderma reesei, with cellobiose and various alcohols. Glucose as a by-product of the reaction was removed using the immobilized yeast system. Among the alkyl glucoside products, the acceptor products of methanol and ethanol were confirmed as methyl ${\beta}$-D-glucopyranoside and ethyl ${\beta}$-D-glucopyranoside via MALDI-TOF MS and enzymatic analysis. Optimal yields of methyl ${\beta}$-glucoside and ethyl ${\beta}$-glucoside were 65.3% (mol/mol) and 59.0% (mol/mol), respectively, based on cellobiose consumed.

• KSBB Journal
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• v.4 no.1
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• pp.21-30
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• 1989

Lauryl alcohol was used as extracting solvent of ethanol, and its toxicity on the free cells or immobilized cells was tested. To increase ethanol productivity, extractive fermentation method combined with ethanol fermentation and ethanol recovery was applied to the immobilized batch and continuous fermenter. As the concentration of LaOH was increased, the lag phase became longer, but specific growth rate did not change greatly. And a cell entrapment technique could protect the yeast cells against both substrate inhibition and solvent toxicity. When the glucose concentration was 400 g/l and the LaOH/fermentation medium ratio was 4, total ethanol productivity increased with the enhancement of LaOH volume, and maximum productivity was 2.75 g/l.hr in the immobilized batch fermentation.

• BMB Reports
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• v.29 no.2
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• pp.169-174
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• 1996

Yeast cytochrome c (cyt c) was modified at cysteine-102 with a thiol-specific spin label and its interaction with liposomes containing acidic phospholipids was studied by electron paramagnetic resonance (EPR) spectroscopy. Association of cyt c with liposomes resulted in a significant reduction in the mobility of the spin label and a fraction of cyt c even seemed to be immobilized. Based on a large spectral change upon binding and the proximity of the spin-label to lysine-86 and -87, we propose these two residues to be the potential binding site at neutral pH. The interaction is electrostatic in nature because the spectral changes were reversed by addition of anions. Dissociation of the bound cyt c by anions, however, became less effective as the lipid/protein ratio increased. This suggests a repulsive lateral interaction among the bound cyt c. Unlabeled cyt c molecules added to preformed cyt c-liposome complex displaced the bound (spin labeled) cyt c and the process was competitive and reversible.

• Journal of Microbiology and Biotechnology
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• v.16 no.3
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• pp.414-422
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• 2006

Before applying PVA bio-beads to practical biological treatment of nitrate-containing wastewater, their characteristics were examined. PVA bio-beads could steadily produce nitrogen gas from nitrate for 28 batches with 0.04 ml/l/h of the maximum gas production rate; however, the maximum gas production rate dropped remarkably thereafter with apparent deformation of beads. Addition of 2.2% solution containing 1% casamino acid, 1% yeast extract, 0.1% mineral solution, and 0.1% vitamin solution to the culture medium resulted in not only recovery of activity of deactivated beads, but also a higher rate of gas production. Calculation of economic benefit for the use of bio-beads in a long-run operation indicated that reactivation of bio-beads by chemicals had economical advantages over packing new bio-beads in the system. The continuously stirred bioreactor exhibited a satisfactory performance at HRT of 20.0 h. With a 9.5 mg $NO_{3}^{-}N/l/h$ nitrate removal rate, nitrate could completely be removed without nitrite accumulation. The use of PVA bio-beads in nitrate removal appears very promising.

• YAKHAK HOEJI
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• v.33 no.6
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• pp.365-371
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• 1989

Microbiological conversion of sterols to 17-ketosteroids has been recognized as a source for commercial preparation of steroidal drugs. In order to develop bacterial strains and process with Brevibacterium lipolyticum IAM 1398 capable of converting cholesterol to 1,4-Androstadiene-3,17-dione (ADD) at about 27% yield, we studied on strain improvement, fermentation condition and whole cell immobilization. By using UV and/or NTG as mutagens, a mutant to convert cholesterol to ADD with higher yield than 60% was selected. Better production of ADD was manifested in the case of maltose used as a supplemental carbon source, and yeast extract or soytone as a nitrogen source. Addition of tween 80 (0.05%) as a surfactant beneficial for increasing the productivity. The optimal initial pH of the medium was 6.5 and optimal culture temperature was $30^{\circ}C$. Whole cell immobilization by using carrageenan, agar, alginate and acrylamide was carried out and the activity of conversion was tested. In the case of carrageenan and agar, immobilized cells were active for at least two cycles of fermentation.