• Microbiology and Biotechnology Letters
• /
• v.20 no.1
• /
• pp.61-67
• /
• 1992

The synthesis of N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester(ZAPM), a precursor of aspartame, from N-benzyloxycarbonyl-L-aspartic acid(Z-Asp) and L-phenylalanine methyl ester hydrochloride(L-PM-HCl) was investigated in ethylacetate-MES buffer two-phase system using thermolysin. In organic two-phase system, the degree of spontaneous hydrolysis of L-PM. HCl was significantly reduced with increasing the volume ratio of organic to aqueous phase. Stability of thermolysin in organic two-phase system was found to be higher than that in MES buffer solution. More than 90% of initial enzyme activity was maintained after 10 days of incubation in case that the volume of organic phase was equal to that of buffer phase, while the half life of thermolysin was about 2 days in aqueous buffer solution. The results of partitioning of substrates and product in organic two-phase system showed that the difference in partition coefficients between substrates and product was maximum at pH 5.5. The optimal pH for 2-APM synthesis in organic two-phase system was found to be 5.5-5.8, which is consistent with the value expected from the partition experiments. As the concentration of substrates was increased the conversion yield of Z-APM was increased with concomitant reduction of L-PMqHC1 hydrolysis. In case that the concentration of L-PM-HCl and Z-Asp were 160 mM and 80 mM respectively, the conversion yield of Z-APM reached 90% after 28 hrs of reaction. The yield obtained at different volume ratio of organic phase compares well with the predicted equilibrium constant in biphasic system.

• Journal of Animal Reproduction and Biotechnology
• /
• v.38 no.2
• /
• pp.70-76
• /
• 2023

Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs. Methods: In vitro-matured oocytes were co-incubated with sperm according to a modified two-step culture system. Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 µL microdroplets (10 gametes/drop) or 500 µL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume. Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence.

• Journal of environmental and Sanitary engineering
• /
• v.14 no.3
• /
• pp.107-115
• /
• 1999

The effectiveness of resting cells of a photosynthetic bacterium, Rhodospirillum rubrum N-1, was investigated on the production of extracellular ${\delta}-aminolevulinic$ acid(ALA). The ALA generating system required 3hr-incubation in the presence of 10mg of resting cells per ml to obtain the maximal yield of extracellular ALA. and also, under this condition the effect of ALA inducers, i.e., 30mM levulinic acid (LA) and L-glutamic acid($C_5$ pathway precursor) was relatively higher than that of produced extracellular ALA($83{\mu}M$). The volume of system and proper cell density appeared to be important factors for the effective production of extracellular ALA.

• Journal of Korean Society for Atmospheric Environment
• /
• v.10 no.E
• /
• pp.325-331
• /
• 1994

Organic extracts of diesel- exhaust particulates were analyzed for mutagenicity using Ames Salmonella typhimurium assay system. An experimental diesel microbus used was drived on the chassis dynamometer according to CVS-75 mode. The samples were taken from the mixed gases in a dilution tunnel. With a high-volume air sampler equipped with double filter holders, particulate matters were collected on a teflon-coated glass fiber filter placed behind a activated carbon filter. After ultrasonic extraction with benzene- ethanol and evaporation to dryness, the residue was dissolved in dimethylsulfoxide. Each sample was tested toward 2 strains, TA100 and TA98 by the pre-incubation method in the absence and presence of S-9mix. Average concentration of diesel- exhaust particulates was about 116.6mg/m$^3$, and 44.1~62.2 ％ to the total weight of particulates, consisted of organic matters. The mutagenicities of diesel- exhaust particulate organic matters were 4,512 and 2,205 revertants/m$^3$ toward TA100 without and with S-9mix, respectively. Those toward TA98 were 13,367 and 3,715 revertants/m$^3$, respectively.

• Archives of Pharmacal Research
• /
• v.10 no.2
• /
• pp.75-79
• /
• 1987

Liposomes were studied as a drug delivery system. Multilamellar vesicles, small unilamellar vesicles and large unilamellar vesicles containing cytarabine were prepared using egg yolk lecithin and cholesterol. Large unilamellar vesicles showed the highest encapsulation efficiency of all and their encapsulation efficiency increased as the buffer volume decreased. Cholesterol increased the stability of liposomal drug products as drug carriers and reduced the permeability of drug across the liposomal membrane. The release rate of cytarabine increased with incubation temperature and decreased with cholesterol incorporation in liposomal membrane. The release mechanism of cytarabine from large unilamellar vesicles in vitro was chiefly due to simple diffusion across the liposomal membrane rather than liposomal rupture.

• Microbiology and Biotechnology Letters
• /
• v.2 no.3
• /
• pp.141-147
• /
• 1974

The cultural conditions for L-glutamate production were investigated using Brevibacterium flavum nov. sp. D2209B, the most productive strain among 5 strains reported in preceeding paper. A temperature of 3$0^{\circ}C$ and a medium volume of 30 ml per 500-flask were selected as standard culture conditions. And the following results were obtained. 1. When the concentration of acetate in the medium was below 30 g per litre, the maximum amount of L-glutamate was accumulated. 2. KH$_2$PO$_4$, MgSO$_4$, FeCI$_3$ and MnCI$_2$ were required for the L-glutamate poduction, but the concentration of those inorganic salts little effected. 3. Signifcant amount of L-glutamate was accutnulated in the limited biotin concentration less than 0.3 ug per litre. 4. The addition of malic acid or succinic acid enhanced the accumulation. 5. The L-glutamate accumulation was related to the incubation time of seed; the amount of L-glutamate accumulated was maximum by inoculating 16-20 hour incubated seed. 6. In the medium containing sufficient amount of biotin for growth, L-glutamate accumulation was stimulated by the addition of penicillin at appropreate time during incubation.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.6
• /
• pp.823-832
• /
• 2021

Mushroom cultivation along with the palm oil industry in Malaysia have contributed to large volumes of accumulated lignocellulosic residues that cause serious environmental pollution when these agroresidues are burned. In this study, we illustrated the utilization of lignocellulolytic enzymes from the spent mushroom substrate of Pleurotus pulmonarius for the hydrolysis of palm oil mill effluent (POME). The hydrolysate was used for the production of biohydrogen gas and enzyme assays were carried out to determine the productivities/activities of lignin peroxidase, laccase, xylanase, endoglucanase and β-glucosidase in spent mushroom substrate. Further, the enzyme cocktails were concentrated for the hydrolysis of POME. Central composite design of response surface methodology was performed to examine the effects of enzyme loading, incubation time and pH on the reducing sugar yield. Productivities of the enzymes for xylanase, laccase, endoglucanase, lignin peroxidase and β-glucosidase were 2.3, 4.1, 14.6, 214.1, and 915.4 U g^-1, respectively. A maximum of 3.75 g/lof reducing sugar was obtained under optimized conditions of 15 h incubation time with 10% enzyme loading (v/v) at a pH of 4.8, which was consistent with the predicted reducing sugar concentration (3.76 g/l). The biohydrogen cumulative volume (302.78 ml H₂.L^-1 POME) and 83.52% biohydrogen gas were recorded using batch fermentation which indicated that the enzymes of spent mushroom substrate can be utilized for hydrolysis of POME.

• Korean Journal of Soil Science and Fertilizer
• /
• v.13 no.2
• /
• pp.57-63
• /
• 1980

The effect of particle size of silicate fertilizer, crushed slag from the steel industry, on the behavior of silicate in soil was investigated through laboratory experiments. The silicate fertilizer was sieved to obtain three fractions of particles, coarser than 10 mesh 20-35 mesh, and finer than 100 mesh. Silicate concentration of the extract obtained by shaking 20 mg of particles, coarser than 10 mesh, 20-35 mesh, and finer than 100 mesh, in 50 ml of distilled water for 4 hours was 0.3, 1.0, and 3.2 ppm respectively. As shaking the mixture of the silicate fertilizer and soil proceeded, silicate concentration of the extract increased, and this increase after 4 hour shaking was attributed mainly to dissolution of soil silicate. When the mixture of soil and the silicate fertilizer was incubated under submerged condition, silicate concentration of the solution decreased for the first 2-4 weeks, thereafter increased with incubation time. During this incubation period, silicate concentration of the solution changed inversely with pH of the solution. After 6-10 weeks, however, both silicate concentration and pH of the solution increased with incubation time. Silicate concentration of the effluent from the 14.5 cm soil column of which top 4.5 cm was packed with the mixture of 30 g of soil and 30 mg of the silicate fertilizer reached maximum at 0.94 pore volumes for the particles of 20-35 mesh and 1.03 pore volumes for the particles finer than 100 mesh, whereas the effluent concentration reached maximum at 0.88 pore volumes for the soil column without the silicate fertilizer treatment. Soil analysis made after water percolation revealed that 1.5 pore volumes of water could leach down large amount of the water soluble silicate but not the sodium acetate extractable silicate, from top 3-6 cm soil layer.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.9
• /
• pp.1278-1285
• /
• 2008

Measurement of gas produced during in vitro fermentation was used to investigate the fermentation characteristics and interactions of rice straw, wheat straw or maize stover mixed with alfalfa or clover at proportions of 100:0, 75:25, 50:50, 25:75 and 0:100, respectively. Cumulative gas production was recorded at 2, 4, 8, 12, 16, 24 and 48 h of incubation, and the Gompertz function was used to describe the kinetics of gas production. In vitro dry matter and organic matter disappearances (IVDMD and IVOMD) were determined after 48 h incubation. The rate of gas production of clover was higher (p<0.05) than that of rice straw, wheat straw, maize stover and alfalfa when straws and hays were incubated separately. Increasing the proportion of alfalfa in the straw-alfalfa mixtures increased (p<0.05) the rates, but not the maximum volume of gas production. However, both rate and the maximum volume of gas production were increased (p<0.01) as the proportions of clover increased in the straw-clover mixtures. The cumulative gas production at 48 h, IVDMD and IVOMD showed no consistent interaction effects between different mixtures of cereal straws and hays. The extent of interactive effects was affected by the types of cereal straw, legume hay and their proportions in the mixture. The appropriate combination for the mixture of rice straw or maize stover with leguminous hays was 75:25 and 25:75, respectively. The better combination occurred at a proportion of 50:50 for the mixture of wheat straw and alfalfa. We conclude that the suitable proportion of low-quality straw and high quality legume hay combination should be considered in the ration formulation system of ruminants according to the extent of positive interactive effects.

• The Korean Journal of Food And Nutrition
• /
• v.9 no.4
• /
• pp.509-515
• /
• 1996

The effects of sour liquid ferments with lactic acid bacteria on the baking properties and qualities of White Pan Bread were studied. The mixed culture of Lactobacillus brevis and Lactobacillus plantarum had higher acid equivalents and lower pH-values than single or mixed culture of other lactic acid bacteria which had been used for traditional sour dough bread. Optimum conditions of the incubation of lactic acid bacteria, which are incubation temperature time and culture medium compositions for lactic fermentation, were also investigated to find out optimum activity for good bread making. The mixed culture of L. brevis and l. plantarum incubated for 24 hours at 3$0^{\circ}C$ had the most optimum activity for bread manufacturing process and the qualities of the products. The addition of sour liquid ferments to the sponge dough effected on fermentation activity of the sponge dough to lower the level of pH to 4.64 and to produce more total titratable acidity(TTA) of 0.545, whereas conventional sponge dough bread had 0.46% of TTA. On comparison with control bread, the bread made with sour liquid ferments was found to have better specific volume, taste, symmetry, especially, organoleptic characteristics due to lactic acid, acetic acid and amino acid produced by lactic acid bacteria. Sour dough bread with liquid ferment was considered to be more effective to the inhibition of staling during storage for 6 days at $25^{\circ}C$ and to have longer shelf-than control.