• Journal of Dairy Science and Biotechnology
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• v.23 no.2
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• pp.115-123
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• 2005

Microcapsules consisting of natural, biodegradable polymers for controlled and/or sustained core release applications are needed. Physicochemical properties of whey proteins suggest that they may be suitable wall materials in developing such microcapsules. The objectives of the research were to develop water-insoluble, whey protein-based microcapsules containing a model water-soluble drug using a chemical cross-linking agent, glutaraldehyde, and to investigate core release from these capsules at simulated physiological conditions. A model water soluble drug, theophylline, was suspended in whey protein isolate (WPI) solution. The suspension was dispersed in a mixture of dichloromethane and hexane containing 1% biomedical polyurethane. Protein matrices were cross-linked with 7.5-30 ml of glutaraldehyde-saturated toluene (GAST) for 1-3 hr. Microcapsules were harvested, washed, dried and analyzed for core retention, microstructure, and core release in enzyme-free simulated gastric fluid (SGF) and simulated intestinal fluid(SIF) at $37^{\circ}C$. A method consisting of double emulsification and heat gelation was also developed to prepare water-insoluble, whey protein-based microcapsules containing anhydrous milkfat (AMF) as a model apolar core. AMF was emulsified into WPI solution (15${\sim}$30%, pH 4.5-7.2) at a proportion of 25${\sim}$50%(w/w, on dry basis). The oil-in-water emulsion was then added and dispersed into corn oil ($50^{\circ}C$) to form an O/W/O double emulsion and then heated at $85^{\circ}C$ for 20 min for gelation of whey protein wall matrix. Effects of emulsion composition and pH on core retention, microstructure, and water-solubility of microcapsules were determined. Overall results suggest that whey proteins can be used in developing microcapsules for controlled and sustained core release applications.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.513-520
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• 2012

This research was planned and executed to evaluate how the composition of Yack-sun tea can affect the health conditions of people who are suffering from diet-related such as being overweight, are obese and have hyperlipidemia, by taking Yack-sun tea in a form of a nutritional supplement with our daily meals. We produced Kangjieum with Lycium chinense Mill., Polygonum multflorum Thunb, Cassia tora L., Crataegus pinnatifida Bge and Salvia miltiorrhiza Bge. Thus, we approach of oriental diet therapy area research of Kangjieum and analysis proximate composition, water soluble antioxidant content. The content(%, dry basis) of total carbohydrate was 60.23%, crude protein was 18.18%, crude ash was 11.36% and crude fat was 10.23% in Kangjieum. Total water soluble antioxidant content was 1.027 ${\mu}g/m{\ell}$ of Kangjieum. We think that scientific and objective evaluation was done on the components of the Kangjieum prescription. This basic data could help guide the application of oriental medicinal resources into other foods and serve as a stepping-stone for use of Kangjieum in the burgeoning field of nutraceutical foods. Last, the scientific effects of oriental medicinal foods developed according to oriental medicinal theory. This theory is believed to be essential for government policy development concerning validation of medicinal effects and assessment, with the aim of fostering systematic development and providing guidance to food development in the interest of national health.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.5
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• pp.637-642
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• 1996

We isolated a marine bacterium, Pseudomonas sp,, which could produce the enzyme of alginate lyase, and cloned the alginate lyase gene in Escherichia coli. The cloned DNA was overexpressed with approximately $50\%$ amount of total proteins. In addition, the expressed proteins were not secreted into the medium, and most of them existed in the cytoplasm by the soluble form, but not observed any inclusion body by TIM. For the optimum enzyme activity, temperature was $20^{\circ}C$, pH was 7.0, and Km and Vmax values of the enzyme were $0.4\%$ and 625 units/mg, respectively.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.155-160
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• 2005

Abstract A direct approach was used to retrieve active lipases from Paenibacillus polymyxa genome databases. Twelve putative lipase genes were tested using a typical lipase sequence rule built on the basis of a consensus sequence of a catalytic triad and oxyanion hole. Among them, six genes satisfied the sequence rule and had similarity (about 25%) with known bacterial lipases. To obtain the six lipase proteins, lipase genes were expressed in E. coli cells and lipolytic activities were measured by using tributyrin plate and pnitrophenyl caproate. One of them, contig 160-26, was expressed as a soluble and active form in E. coli cell. After purifying on Ni-NTA column, its detailed biochemical properties were characterized. It had a maximum hydrolytic activity at $30^{\circ}C$ and pH 7- 8, and was stable up to $40^{\circ}C$ and in the range of pH 5- 8. It most rapidly hydrolyzed pNPC$_6$ among various PNPesters. The other contigs were expressed more or less as soluble forms, although no lipolytic activities were detected. As they have many conserved regions with lipase 160-26 as well as other bacterial lipases throughout their equence, they are suggested as true lipase genes.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.542-549
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• 2010

In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

• Journal of environmental and Sanitary engineering
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• v.13 no.1
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• pp.16-25
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• 1998

Laboratory-scale experiment using anaerobic fluidized bed reactor was carried out to investigate the prehydrolysis step with caustic soda on the treatment efficiency of anaerobic sludge treatment, since the overall rate-limiting step for the complete anaerobic digestion of sludge was the hydrolysis step by extracellular bacterial enzymes of insoluble polymeric molecules. Reactors received a sludge which had not been pretreated, a 50-50 mixture of pretreated and untreated sludge, and the fully pretreated sludge. Hydraulic retention time of 10, 5, 2.5 days and 1 day were applied with an respective equivalent organic loading rate of 1.17, 2.23, 4.17, 11.24 gCOD/L/d. Reactor with the untreated sludge did not archieve adequate digestion even at the HRT of 5 days, and reactor, which received the 50-50 mixture, operated well at the HRT of 5 days, but began to show signs of unstable digestion at the HRT of 2.5 days. While, reactor, which was fed the hydrolyzed sludge, operated reasonably well at the 2.5 days, but was showing somewhat decrease in removal efficiencies. Results, therefore, have substantiated that the limiting reaction in the anaerobic treatment process is hydrolysis. The soluble COD did not significantly accumulate in the reactor as organic acid form, even when they were highly stressed. It was believed that this resistance to a build-up of organic acids and soluble COD behavior was mainly due to the maintenance of the methane bacteria in the fixed-film system which prevents washout as the organic loading increased. The anaerobic fluidized bed reactor was therefore effective for the digestion of waste activated sludge at short HRT.

• Biomolecules & Therapeutics
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• v.19 no.1
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• pp.70-76
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• 2011

Bone remodeling is a dynamic process involving a constant balance between osteoclast-induced bone resorption and osteoblast-induced bone formation. Osteoclasts play a crucial homeostatic role in skeletal modeling and remodeling, and destroy bone in many pathological conditions. Previously, we reported that the hexane soluble fraction of Ficus carica inhibited osteoclast differentiation. Poly unsaturated fatty acids, such as ethyl docosahexaenoate (E-DHA), docosahexaenoic acid (DHA), cis-11,14-eicosadienoic acid (EDA) and eicosapentaenoic acid (EPA), were identified from the hexane soluble fraction of Ficus carica. Among them, E-DHA most potently inhibited osteoclastogenesis in RAW264.7 cells. E-DHA reduced the activities of JNK and NF-$\kappa}B$. E-DHA suppressed the expression of c-Fos and nuclear factor of activated T cells c1 (NFATc1). Interestingly, DHA increased the activity of alkaline phosphatase and expression of bone morphogenetic protein 2 (BMP2) more than E-DHA in MC3T3-E1 cells, suggesting that DHA may induce osteoblast differentiation. The data suggests that a combination of E-DHA and DHA has potential use in the treatment of diseases involving abnormal bone lysis, such as osteoporosis, rheumatoid arthritis and periodontal bone erosion.

• Journal of Pharmaceutical Investigation
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• v.6 no.3
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• pp.48-57
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• 1976

An enhancement in the dissolution rate of the drug should facilitate its GI absorption if the absorption process is dissolution rate limited. One of the need for the techniques that can potentially enhance the dissolution rate and extent of absorption of hydrophobic drugs is the formation of coprecipitates with pharmacologically inert, polymeric materials. The physicochemical modification offers the advantage of possibly enabling one to administer the drug orally in a form from which it is most available for GI absorption. Several $investigation^{1-15)}$ demonstrated that the formation of solid dispersions or coprecipitates of relatively water-insoluble drugs with various pharmacologically inert carriers can increase singnificantly their in vitro dissolution rates. However, little information is available in the literature related to the dissolution rate patterns of furosemide, a water-insoluble diurectices, with respect to the sort of copolymer and the ratio of coprecipitates as a function of time, respectively. The purpose of the present investigation was to ascertain, the general applicability of the copolymers to use fore more fast, enhanced dissolution techniques of furosemide. To accomplish the need for enhancement in the dissolution rate of furosemide, varying ratio coprecipitates with different water-soluble polymers, such as polyvinylpyrrolidone (PVP), polyethylene glycol 4000(PEG 4000), and polyethylene glycol 6000 (PEG 6000), were quantitatively studied by comparing their dissolution characteristics of furosemide. The dissolution patterns of pure furosemide, varying ratio furosemide-PVP coprecipitates, (1:2, 1:5, and 1:9(w/w)), furosemide-PEG 4000 coprecipitates (1:4, 1:9, and 1:19(w/w), furosemide-PEG 6000 coprecipitates(1:4, 1:9, and 1:19(w/w)), and the same ratio physical mixtures, respectively, were compared by the amount dissolved as a function of time.

• Applied Chemistry for Engineering
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• v.18 no.6
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• pp.607-611
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• 2007

To improve transfection efficiency, we prepared histidine-low molecular weight water-soluble chitosan (LMWSC) having the potential to form complex with DNA as a cationic polymer. Histidine-LMWSC was synthesized by the esterification reaction and removing phthaloyl group. The histidine-LMWSC was characterized using FT-IR, $^1H$ NMR spectra. Histidine-LMWSC was complexed with plasmid DNA (pDNA) in various polymer/DNA (N/P) weight ratios, and the complex was identified using gel retardation assay. The particle sizes of the hisitidine-LMWSC/DNA complexes were measured on a DLS instrument by fixing the histidine-LMWSC/DNA weight ratio of 10/1. Owing to the utilization of a large excess amount of cationic LMWSC against anionic DNA, the particle size of histidine-LMWSC/DNA complexes was in the range of 100~200 nm. Therefore, histidine-LMWSC will be useful in the development of gene carriers.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.2
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• pp.67-75
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• 2002

The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.