• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.34-38
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• 2003

For the production of $B^{30}-homoserine$ insulin analog as a novel anti-diabetic drug, the fermentative study was attempted for the maximal gene expression of HTS-fused $B^{30}-homoserine$ insulin precursor in the recombinant Escherichia coli cells. In a batch fermentation, the maximal production of insulin precursor as much as 38.95 mg/L-h, which occupied more than 12.8% of total cell protein. was achieved when the gene expression was induced by 0.5 mM IPTG at the middle logarithmic growth phase. The HTS-fused $B^{30}-homoserine$ insulin precursor was recovered from a batch culture through the processes of cell harvest, collection of insoluble fraction after sonication and purification by nickel affinity column chromatography. The isolated insulin precursor was 14 mg/L with a recovery yield of 35.9% of expressed gene product. The insulin A and B chain mixture was recovered after the insulin precursor was subjected to CNBr cleavage and purified by nickel affinity column chromatography. The isolated insulin chains were then sulfitolyzed with sodium thiosulfat and sodium tetrathionate, and reconstituted to insulin analog with ${\beta}-mercaptoethanol$, followed by purification with CM-Sepharose C-25 column chromatography.

• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1382-1388
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• 2014

Proteus vulgaris K80 lipase was expressed in Escherichia coli BL21 (DE3) cells and immobilized on amine-terminated magnetic microparticles (Mag-MPs). The immobilization yield and activity retention were 84.15% and 7.87%, respectively. A homology model of lipase K80 was constructed using P. mirabilis lipase as the template. Many lysine residues were located on the protein surface, remote from active sites. The biochemical characteristics of immobilized lipase K80 were compared with the soluble free form of lipase K80. The optimum temperature of K80-Mag-MPs was $60^{\circ}C$, which was $20^{\circ}C$ higher than that of the soluble form. K80-Mag-MPs also tended to be more stable than the soluble form at elevated temperatures and a broad range of pH. K80-Mag-MP maintained its stable form at up to $40^{\circ}C$ and in a pH range of 5.0-10.0, whereas soluble K80 maintained its activity up to $35^{\circ}C$ and pH 6.0-10.0. K80-Mag-MPs had broader substrate specificity compared with that of soluble K80. K80-Mag-MPs showed about 80% residual relative activity after five recovery trials. These results indicate the potential benefit of K80-Mag-MPs as a biocatalyst in various industries.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.9
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• pp.1369-1373
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• 2003

ACE (Angiotensin-I converting enzyme) inhibitory peptides derived from foods are thought to suppress high blood pressure by inhibiting ACE. We tried to make efficient production of the ACE inhibitory hydrolysate from egg albumen. A hydrolysate digested by neutrase presented the highest ACE inhibitory activity ($IC_50\;value=256.35{\mu}g/ml$) and the proper proteolysis was occurred by 1.0% enzyme addition and 4 h incubation at $47^{\circ}C$. Antihypertensive effect of neutrase hydrolysate was investigated in spontaneously hypertensive rats (SHR, n=5). Systolic blood pressure (SBP) was decrease by 6.88% (-14.14 mmHg, p<0.05) at 3 h after oral administration of 300 mg/kg body weight, and by 13.33% (-27.72 mmHg, p<0.05) by emulsified hydrolysate. These results showed that it is very effective to utilize egg albumen as a protein source for the production of ACE inhibitory peptides. However, further studies are required to investigate the methods to increase recovery yield and the isolation of active peptide is necessary for determining its sequence responsible for ACE inhibitory activity.

• Environmental Engineering Research
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• v.19 no.3
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• pp.185-195
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• 2014

Despite the enormous technical and economic efforts to improve environmental conditions, currently about 40% of the global population (or 2 billion people) are still lack access to safe water supply and adequate sanitation facilities. Pollution problems and transmission of water- related diseases will continue to proliferate. The rapid population growth and industrialization will lead to a reduction of arable land, thus exacerbating the food shortage problems and threatening environmental sustainability. Natural systems in this context are a transdisciplinary approach which employs the activities of microbes, soil and/or plants in waste stabilisation and resource recovery without the aid of mechanical or energy-intensive equipments. Examples of these natural systems are: waste stabilisation ponds, aquatic weed ponds, constructed wetlands and land treatment processes. Although they require relatively large land areas, the natural systems could achieve a high degree of waste stabilisation and at the same time, yield potentials for waste recycling through the production of algal protein, fish, crops, and plant biomass. Because of the complex interactions occurring in the natural systems, the existing design procedures are based mainly on empirical or field experience approaches. An integrated kinetic model encompassing the activities of both suspended and biofilm bacteria and some important engineering parameters has been developed which could predict the organic matter degradation in the natural systems satisfactorily.

• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.658-665
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• 1995

Low yield of a thermostable pectinesterase(TSPE) from citrus fruits has made its detailed study extremely tedious and difficult; therefore, maximizing TSPE extraction is desirable. It is assumed that TSPE is bound to the cell components via ionic linkage and covalent bonds. Therefore, in this study, variations in extraction time, pH, NaCl concentration and commercial enzyme preparations were used to increase the yield of TSPE from Valencia orange. The largest recovery of TSPE, obtained by heating extracted pectinesterase(PE) at $70^{\circ}C\;for\;5{\sim}10$ minute, was achieved using actate buffer(pH 4.14) with 1 M NaCl and 0.2% $Cytolase^{TM}$ 104(a mixture of cellulase, hemicellulase and pectinase; Genecor, Inc). The two aquous phase partitioning with 5.0% Triton X-114 could be used as a tool for separation of thermolabile pectinesterase(TLPE) and TSPE from crude PE. Also, water extraction and $0{\sim}0.3$ ammonium sulfate fractionation could be used for removing non-pectinesterase protein.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.196-203
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• 2002

A novel process for urokinase purification was studied using p-aminobenzamidine as the ligand and sepharose 4B as the matrix. The adsorption, washing, and elution conditions were optimized by an unusual method. An adsorption buffer containing 2.5 M NaCl and $1\%$ Tween 80 facilitated the adsorption of urokinase on the affinity media and prevented contaminants from binding to the p-aminobenzamidine affinity gel. It was found that $5\%$ Tween 80 removed most of the contaminants from the affinity column. A 0.2 M glycine elution buffer containing 0.5 M NaCl (pH 3.0) was found to have a strong elution ability with a high recovery and purity of urokinase. A crude urokinase material of231 IU/mg protein from human urine was purified to 124,300 IU/mg protein with a purification factor of 538 and yield of $86.7\%$. As a result, a high purity urokinase was obtained with only a single affinity chromatography step. The purification process was successfully scaled-up to a 2-1 chromatography column. The resulting urokinase eluate could be directly lyophilized, thereby complying with Chinese pharmacopoeia (1995 version) standards.

• Journal of The Korean Society of Grassland and Forage Science
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• v.5 no.3
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• pp.187-194
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• 1985

This field experiment was carried out to determine the effects of thirteen different fertilizer levels of nitrogen(N), phosphorus($P_2O_5$) and potassium($K_2O$) on the content of crude protein, crude fiber, mineral constituents of product and tree growth forest pasture with 40-50% shading. The experiment was arranged as a randomized block design and performed in the suburban forest of Suweon in 1984. The results obtained are summarized as follows: 1. Crude protein content and total protein yield were higher in the plot of 28 and 42kg $N/_{10a}$, regardless of $P_2O_5\;and\;K_2O$ level, while those were the lowest in zero fertilizer and N-zero fertilizer plots. 2. The contents of lignin and silica were significantly low in the high N fertilizer level, and the contents of NDF, ADF, cellulose and hemicellulose were not affected by different fertilizer levels. However, the content of crude fiber tended to be low with high N, regardless of $P_2O_5\;and\;K_2O$. 3. The contents of N,K and $SiO_2$ of grasses were influenced by different fertilizer levels. However, those of P,Ca,Ma and Na showed little differences. 4. The recovery percentage of NPK was higher in the plot of standard fertilizer level with 28-20-24 kg/10a, and higher recovery percentage was observed in $K_2O$, followed by N and $P_2O_5$ in that order. 5. The growth of tree was increased as the level of N fertilizer was increased, but no such trend was noted by $P_2O_5\;and\;K_2O$ levels. 6. Crude protein, crude fiber, some mineral contituents of grasses, and growth of tree were influenced by N level, regardless of $P_2O_5\;and\;K_2O$. And the optimum fertilizer level of $N-P_2O_5-K_2O$ seemed to be 28-20-24 kg/10a for the production of grasses with higher quality and more yield in the forest.

• Korean Chemical Engineering Research
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• v.43 no.2
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• pp.187-201
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• 2005

Bioprocessing technologies utilizing 'biorecognition' between a solid matrix and a protein is being widely experimented as a means to replacing the conventional, solution-based technology. Frequently the matrices are chromatographic resins with specific functional groups exposed outside. Since the reactions of and interactions with the proteins occur as they are attached to the solid matrix, this 'solid-phase' processing has distinct advantages over the solution-phase technology. Solid-phase refolding of inclusion body proteins uses ion exchange resins to adsorb denaturant-dissolved inclusion body. As the denaturant is slowly removed from the micromoiety around the protein, it is refolded into a native, three-dimensional structure. Once the refolding is complete, the folded protein can be eluted by a conventional elution technique such as the salt-gradient. This concept was successfully extended to 'EBA (expanded bed adsorption)-mediated refolding,' in which the denaturant-dissolved inclusion body in whole cell homogenate is adsorbed to a Streamline resin while cell debris and other impurity proteins are removed by the EBA action. The adsorbed protein follows the same refolding steps. This solid-phase refolding process shows the potential to improve the refolding yield, reduce the number of processing steps and the processing volume and time, and thus improve the overall process economics significantly. In this paper, the experimental results of the solid-phase refolding technology applied to several biopharmaceutical proteins of various types are presented.

• Korean Journal of Food Science and Technology
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• v.22 no.3
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• pp.363-368
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• 1990

As a work on the investigation of the processing aptitude of the Korean soybean cultivars, the soybean curd was made from 19 Korean soybean cultivars on a laboratory scale. The yield, proximate composition and rheological and organoleptic property of the resultant soygean curd were analyzed. The yields of soybean curd were ranged from 262.33% to 311.63%, being positively correlated with the soluble protein contents of the recommended soybean cultivars (r=0.9621). The recovery rate of protein in Baekun and Yeunha to the soybean curd was 72.94%, 70.62%, respectively. More than 80% of lipid in soybeans of Yeunha, Dankyung, Danyeob was transferred to the soybean curd. Lightness of the soybean curd made from Baekun, Kwanggyo, Saeal, Hill was higher than that of the others. The mechanical hardness of the soybean curd was strongly correlated with the final moisture content of the soybean curd (r =-0.8312), but not with the protein content of the soybean (r=0.1848). In the organoleptic test for color, texture, flavor and overall preference of the resultant soybean curd, the quality of Dankyung, Namcheon, S-138, Baekun was evaluated highly.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.49-57
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• 1999

Enzymatic hydrolysis with tuna pyloric caeca crude enzyme(TPCCE) was performed to recover a protein hydrolysate from hoki frame, fish processing by-product. Optimum hydrolytic conditions were pH 10.0, temperature $50^{\circ}C$, and incubation time 12 hrs, and then the degree of hydrolysis was about 60%. The yield of the hydrolysate from hoki frame by enzymatic hydrolysis was approximately 77% on a dry weight basis. The prepared protein hydrolysates were also fractionated through a series of 30, 10, 5 and 1 kDa molecular weight cut-off (MWCO) membranes in order to investigate the effect of their functionalities according to the difference of their molecular size. As the result of studying functionalities of the hydrolysates, 1 K hydrolysate showed the highest solubility over all pHs, and 30 and 10 K hydrolysate showed more excellent emulsifying property and whippability than the other hydrolysates.