Experiments were carried out to optimize the fermentation conditions for the production of erythritol by salt-tolerant mutant, Candida magnoliae M26. The optimum conditions of erythritol production showed a 1.0 vvm aeration and 500 rpm agitation at 28 with an initial medium pH of 7.0. The pH control during the fermentation did not improve the erythritol yield and productivity. The maximum erythritol concentration of 143.3 g/L was obtained with 57% yield and 0.70 g/L-h productivity from 250 g/L of glucose and 5 g/L of yeast extract under an optimum fermentation conditions. The medium containing 0.5 M KCl or 0.5 M NaCl enhanced the production of erythritol and glycerol. However, glycerol production increased and erythrtiol production decreased by increasing the concentration of NaCl or KCl.

The changes in the levels of total soluble protein and some free amino acids were investigated in germinating brown rice. Nongerminated (N) brown rice was germinated for 72 hrs by applying following solutions: 1) distilled water (W), 2) 50 ppm chitosan in 5 mM lactic acid (CL), and 4) 50 ppm chitosan in 5 mM glutamic acid (CG). The level of total soluble protein was higher in the N extract than those of W, CL and CG. Alanine levels were enhanced and aspartic acid levels were decreased significantly in the germinated brown rice, highest increases of alanine were found in the CG germinated brown rice. The levels of serine, decreased during germination in solutions W and CL, were increased significantly by germination in CG solution. The levels of essential amino acids, such as Iysine, isoleucine and methionine were also increased significantly by germination in CG solution. Our results show that the germination of brown rice with CG solution can significantly increase the levels of alanine and some other essential amino acids and can restore the serine level.

The present study was designed to investigate the effects of Stachys Sieboldii MIQ as a new natural antitumor agent or immunomodulator. To obtain the above objectives, Stachys sieboldii MIQ was extracted with ethanol. Stachys sieboldii MIQ accelerated mouse spleen cell growth, but inhibited FM3A/S°-cell growth. However, no significant difference was found for CD4+ / CD8+ cells. The growth rates of CD4+ and CD8+ T cells were accelerated more than those normal mouse group. Stachys sieboldii MIQ fed mice showed a significant enhancement of IL-2 receptor expression, increased numbers of CD4+ T cells, and CD8+ T cells. Stachys sieboldii MIQ also stimulated the production of NO by peritoneal macrophages and the production of NO by and the growth of mouse spleen cells. On the other hand, lung localization of Bl6Fl0 melanoma cells was inhibited by ethanol extract of Stachys sieboldii MIQ. These results show that Stachys sieboldii MIQ is a useful new functional antitumor agent or immunomodulator.

The optimum culture conditions for tropane alkaloid production in hairy root cultures of Korea native Scopolia paviflora Nak. were investigated. Hairy root was induced from the rhizome of the mother plant on B5 medium containing 1.0 mg/L IBA. Among the culture media examined, 1/2 B5 medium was the best for tropane alkaloid production, whereas the growth of hairy root increased in SH medium. The best result on the growth of hairy root was obtained in 1.0 mg/L NAA, and tropane alkaloid production was obtained in plant growth regulator-free medium. Of the carbone sources tested, 3% sucrose promoted the growth of hairy root, whereas 5% sucrose increased tropane alkaloid production. Optimum inoculum densities for root growth and tropane alkaloid production were 0.5 g and 1 g, respectively. The addition of XAD resins (1 % w/v) to hairy root cultures led to increases in tropans alkaloid production, and the release of alkaloid into the medium and its adsorption by the resin accounted for about 50 to 80％ of total production. It is concluded that optimized culture conditions and the addition of XAD resins could be used in the development of a bioprocess for tropane alkaloid production in hairy root cultures of S. paviflora Nak.

In this study, we attempted to increase the survivability of bifidobacteria in simulated gastric juices and bile salts after cell entrapment with alginate and various food additives, such as erythritol, isomalt, palatinose, skim milk, xanthan gum, isomalto-oligosaccharide, fructo-oligosaccharide, galacto-oligosaccharide, pectin, and mono-sodium glutamate. Additionaly, the stability of bifidobacteria during storage was investigated by measuring survival rate at different temperatures, i.e. at 4, 25 and -20. Bifidobacteria were immobilized in alginate beads and the survival rate was monitored. It was found that bifidobacieria entrapped with 2.5％, alginate showed the highest survival rate at 12%. After addition of the various protective agents, erythritol(1％) showed the best protective efficiency with a survival rate of 56.0% among the additives tested when exposed to simulated gastric juices for 3 h. Immobilized cells suspended in 5% skim milk and stored at 4 survived significantly more than cells stored at 25 and -20. Consequently, the study shows that the survival rate of bifidobacteria immobilized in combination with 2.5% alginate beads and 1% erythritol may be signifcantly increased in simulated gastric juices and bile salts.

To screening of antimicrobial activity, 95% ethanol and hot water extracts of roots, fruits, leaves, radix and stems of 50 species of traditional herbal medicines were examined. For their growth inhibitory effects on two food-born microorganisms, S. aureus and E. coli O157:H7, by the paper disc diffusion method and the minimum inhibitory concentration(MIC) test. Moutan radicis Cortex and Achyranthis Radix showed the highest inhibitory activities on both S. aureus and E. coli O157:H7. The Inhibition zones of Moutan radicis Cortex on S. aureus and E. coii O157:H7 were 22 mm and 24 mm respectively, and the corresponding inhibition zone of Achyranthis Radix were 23 mm and 22 mm. The MIC or Achyranthis Radix on S. aureus was 156.25 g/mL, and the MIC or Achyranthis Radix and Moutan radicis Cortexas on E. coli O157:H7 were 625 g/mL and 312.5 g/mL, respectively. Their antimicrobial activities in ethanolic extracts were significantly higher than in hot water extracts. In the various solvent fractions prepared from ethanol extract, the ethyl acetate fraction of Achyranthis Radix and the CHCl fraction of Moutan radicis Cortexas showed strongest activity.

The enantioselective esterification of racemic ibuprofen catalyzed by a Candida rugosa lipase was studied according to reaction conditions such as a lipase concentration, reaction temperature, alcohol chain length and alcohol concentration. The S-(+)-ibuprofen alkyl esters prepared were converted to S-(+)-ibuprofen by hydrolysis with sulfuric acid as a catalyst. High conversions in the esterifications were obtained at 60 and an equimolar ratio of octanol to ibuprofen. The initial reaction rate of the esterification decreased with increasing octanol concentration. Conversion and initial reaction rate increased with increasing alcohol chain length. Values of enantiomeric excess(ee) according to esterification reaction conditions did not change below 60. On the other hand, values of conversion and ee for the chemical hydrolysis of S-(+)-ibuprofen alkyl esters were independent of alcohol alkyl chain length. Optical resolution of racemic ibuprofen was achieved by lipase catalyzed esterification and chemical hydrolysis. The separation method provided a high yield and enantioselectivity for the production of S-(+)-ibuprofen from racemic ibuprofen.

The hydrolysis of old book(Hanji) was performed using endoglucanase Ⅰ(endo Ⅰ), and exoglucanase II(exe II) and their mixtures purified from Trichoderma viride cellulase. The optimum degradation of old book(Hanji) with endo Ⅰ, exo II and endo-exo mixture(Ⅰ：Ⅰ) were exhibited at pH 4.5, 5.5, 5.0, respectively. Maximum degradations using endo Ⅰ, exo II and endo-exo mixture(Ⅰ：Ⅰ) occurred at 50. The yield decreased an increasing the enzyme concentration. Especially, the yield was lowest for treatment with the endo Ⅰ-exo II mixture(Ⅰ：Ⅰ), which may be regarded as being due to a synergistic action of the cellulase components. Physical strength increased with increasing exo II concentration, and decreased with increasing concentration of endoglucanase Ⅰ. These results indicated that the degradation of old book(Hanji) depends largely upon the action of endoglucanase. Therefore, the most effective method of conserving paper cultural properties is to repress the action of endoglucanase.

Three different strains of Aureobasidium pullulans were grown in batch cultures to compare their abilities of enzyme production. It was found that specific enzyme activity was the highest with strain ATCC 9348 and the enzyme production was closely coupled to growth. Studies on morphology during the growth of A. pullulans revealed that mycelia cells were dominant at the initial stages of growth. However, yeast-like cells and chlamydospores were dominant in the latter stages of batch culture. The pattern of morphological changes during the growth period was not affected by pH. However, it appears that the ratio of intra- to extracellular enzyme activity tended to increase with fermentation time irrespective of the pH employed, suggesting that the secretion efficiency of intracellular enzyme to broth likely depends on cell morphology Using molasses as a cheap source of sucrose, enzymatic production of fructo-oligosaccharides as a feed additive with A. pullulans cells could be achieved successfully at 55 and pH 5.5.

A COG(clusters of orthologous groups of proteins) algorithm was used to detect conserved genes within Proteobacteria and to figure out their relationships. Restricting comparison to the sequences of 42 procaryotes, 33 eubacteria and 16 Proteobacteria, the number of conserved genes was increased. All analyzed procaryotes shared 75 COGs. COG0195, COG0358 and COG0528 were only represented by the 42 procaryotes. Sixtyfour COGs were added as conserved genes in 33 eubacteria. Each Proteobacteria group has a unique repertoire of COGs. Metabolic COGs were more diverse in the beta Proteobacteria group than in the other groups. These results could be used to determine the origins and the evolutionary relationships of Proteobacteria. The possibilities of detecting new biological molecules is high in phylogenetically related organisms, hence the identification of useful proteins by using this algorithm is possible.

In lactic acid fermentation, the end product inhibition by lactic acid causes several problems. The most important of which are low lactate formation rate and its recovery from fermentation broth. To overcome these problems, extractive lactic acid fermentation was carried out in a bioreactor, which was connected to a column packed with anion exchange resin (Amberlite IRA-400, 250 g). The system was started as a batch process, and then the separation process was started when the lactic acid concentration reached 10 g/L, 20 g/L or 30 g/L. In each case, total lactic acid concentration was reached to 48.6, 53.6, 52.6 g/L with its productivity of 1.2 g/L h, 1.6 g/L h, and 1.3 g/L h, respectively Especially, in the case of the 20 g/L recycling-initiation process, extractive fermentation reduced tie fermentation time (17 hrs) by 34% in comparison with the conventional batch process. The direct consequence of this time reduction was shown by a 1.8 fold increase in overall lactic acid productivity.

Recombinant E. coli DH5 /pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD value of 0.6-0.7. ADout 10-10 cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

In an attempt to produce recombinant proteins using the pollen enriched in some plant species, a 1.7 kb DNA encoding urease subunit B (UreB) amplified by PCR from Helicobacter pylori urease gene cluster in pH808 plasmid was cloned to be expressed under CaMV35S promoter in lily (Lilium longiflorum) pollen tubes elongated in vitro. Lily pollen at early germinating stage was transformed with the ureB DNA using Agrobacterium via vacuum infiltration and, incubated for a full pollen tube growth 16 - 24 h in the dark in the presence of kanamycin. DNA integration and expression in the transgenic pollen were analyzed by the standard molecular techniques and the results suggest that the pollen in vitro may be employed as a protein factory in a disposable fashion.

In an effort to use plant biotechnology for the production of biopharmaceuticals, pollens collected from lily (Lilium longiflorum) were grown in vitro and transformed with a PCR-amplified 1.7 kb cDNA encoding human tissue plasminogen activator (tPA) using Agrobacterium via a vacuum infiltration process. Western blotting showed that transgenic lily pollen tubes selected on kanamycin for 16 hrs expressed a tPA protein with a size similar to the human standard, suggesting their possible use as a disposable host for rapid foreign protein production.

Lipomyces starkeyi KSM 22 produces dextranase and amylase (DXAMase). The addition of 0.02％ (w/v) 2-deoxy-D-glucose or 0.5% (w/v) glycerol into a 1% (w/v) starch medium increased the final activity of DXAMase produced 2.5 fold or 2.4 fold, respectively, compared to that of a 1% (w/v) starch medium. This activity was similar to that produced with 1% (w/v) dextran. The stability of purified OXAMase at 40 for 3 weeks was tested using the various enzyme stabilizers. With the addition of 25% (v/v) glycerol, 90.9% of initial activity was left after 3 weeks. For practical use, the addition of 1% (v/v) glycerol with 50 mM of CaCl or KHPOwas adequate and maintained 73.4% of the initial activity under the test conditions used.