• Journal of Microbiology and Biotechnology
• /
• v.10 no.5
• /
• pp.663-669
• /
• 2000

The fur (ferric uptake regulation) expression vector pMON2064 was modified to produce a Fur-fusion expression vector. A kinker site, factor Xa cleavage site, and several restriction endonuclease sites were introduced to facilitate easy cloning and isolating of the fusion protein. The resulting fusion expression vector, pMONSTER, was then used to make fusion expression vector, pMONSTER, was then used to make fusion proteins with $\beta$-galactosidase and the protease of the human immunodeficiency virus type 1 (HIV-1 PR). Strain SW4020 harboring the Fur $\beta$-galactosidase fusion vector produced blue colonies on a 5-bromo-4-chloro-3-indolyl-$\beta$-D-galactoside plate and the resulting 133 kDa fusion protein reacted with an anti-Fur antibody. The strain harboring the Fur-HIV-1 PR fusion vector produced a 29 kDa fusion protein, which also reacted with an anti-Fur antibody. The Fur-HIV-1 PR fusion protein was purified by a single column application that was designed to isolate the Fur protein. The purified Fur-HIV-1 PR fusion protein digested with factor Xa cleaved a recombinant Gag protein to release smaller fragments, including a p24 capsid protein. The Fur-HIV-1 PR fusion protein itself did not exhibit any proteolytic activity.

• Journal of Pharmaceutical Investigation
• /
• v.36 no.2
• /
• pp.115-121
• /
• 2006

The aim of this study was to develop new protein/peptide depot system instead of poly(DL-lactic acid-coglycolic acid) (PLGA) microspheres. Pullulan was chemically modified by the addition of acetic anhydride (pullulan acetate; PA) and then investigated as new depot system for protein/peptide delivery. PA microspheres (PAM) with lysozyme as a model protein were prepared by w/o/w double emulsion method. The microspheres had a mean size of 10-50 mm with a spherical shape. The size distributions reduced with increasing the degree of acetylation. The loading efficiency of lysozyme was also increased. Lysozyme aggregation behavior in the microsphere was monitored to estimate the change of protein stability during preparation step. The ratios of protein aggregation in PAMs are lower than that of PLGA microsphere, in particular, PA 5 showed lowest as about 16%. The result indicated that the increase of acetylation suppressed the aggregation of protein. The release profiles of lysozyme from PAMs were significantly different. High acetylation effectively improved lysozyme release kinetics by reducing initial burst release and extending continuous release over a period of time. To check the effect of preservation for structural stability of lysozyme, the activity of lysozyme released from PA 5 was also observed. The activity of lysozyme was maintained almost 100% for 25 day. Therefore, PAM may become to a useful carrier for delivery of protein/peptide drugs, if it will be supported by biocompatibility and biodegradability results.

• Journal of Microbiology and Biotechnology
• /
• v.12 no.1
• /
• pp.65-70
• /
• 2002

A TGF-${\beta}1$/GFP monomeric fusion protein was cloned from pPK9A and pGFP-Cl plasmid by PCR amplification. The fusion protein was expressed in a $Bac-To-Bac^{TM}$ baculovirus expression system. A 45 kDa fusion protein was purified using an Ni-NTA column with 300 mM imidazol from a cell lysate infected with recombinant viruses for 72 h post-infection. The fusion protein cross-reacted with the commercial $TGF-{\beta}1$ polyclonal Ab as well as Ab raised against a precursor, monomeric $TGF-{\beta}1$, and GFP. The binding activity of the fusion protein with a $TGF-{\beta}1$ receptor was examined. Fluorescence was observed in Mv1Lu cells, yet not in insect cells treated with the fusion protein. No fluorescence was detected in Mv1Lu cells incubated with the fusion protein treated with Ab prior to the binding reaction, or with GFP alone, thereby indicating that the binding of the fusion protein was specific to $TGF-{\beta}1$ with a receptor.

• Journal of Nutrition and Health
• /
• v.29 no.8
• /
• pp.874-880
• /
• 1996

The effects of an anabolic steroid, nandrolone phenylpropionate(NPP), on body weight gain and body protein, and muscle protein metabolism were inestigated in adrenocorticotrophic hormone(ACTH)-treated male and female rats. Daily injections of 100ug/day of ACTH for 7-8 days caused a cessation of growth in females and a net loss of body weight in males which were associated with significant reductions in body protein content. However, food intake was not affected by ACTH in either sex. The weight, protein content and fractional rate of protein synthesis, measured in vivo, of gastrocnemius muscle were all significantly reduced in both sexes. NPP at a dose of 4mg/kg body weight prevented the reduction in body weight gain in ACTH-treate females but not in males. However, boy protein content was increased by NPP in both sexes which was associated with increases in the weight, protein content and fractional rate of protein synthesis of gastrocnemius muscle. ACTH treatment caused a marked increase in plasma concentrations of corticosterone in both sexes. NPP suppressed much of the increases in corticosterone concentrations in both sexes. The results of the present study suggest that NPP exerts at least part of its anabolic effect by reducing plasma concentrations of catabolic glucocorticoid hormones, through suppressing the response of the adrenals to ACTH.

• Journal of Nutrition and Health
• /
• v.19 no.1
• /
• pp.16-22
• /
• 1986

This study was designed to investigate the nutritional effect of mugowrt powder supple mentation to rice diets with different protein levels. Thirty female albino rats weighing 39-43g were adopted for the feeding trial for 4 weeks. The different 5 kind of experimental diets were performed . Control diet was commercially available forage for rats, experimental diet (I-C) highly milled rice, (Ⅰ) highly milled rice 95% and mugwort powder 5%, (Ⅱ-C)highly milled rice 95% and milk casein 5%, and (Ⅱ) highly milled rice 90%, milk casein 5 % and mugwort powder 5%. Growth rate was remarkably high in the dietary group fed on highly milled rice supplemented with 5% mugwort powder (protein 8%) (P<0.05), but it showed the tendency to be rather low in the group fed on highly milled rice supplemented with 5% casein and 5% mugwort powder (protein 12%). Food efficiency as well as protein efficiency appeared similar to the growth rate. Hematodcrit level demonstrated the same tendency as growth rate, but hemoglobin content was observed to increase by diets supplemented with increasing amount of mugwort regardless of protein level. Each nutrient intake was increased by adding mugwort powder to diets after a week's feeding, but it was increased by 8% protein diet, and decreased by 12% protein diet in 3 weeks as well as 4 weeks after feeding . The absorption rate of carbohydrate and protein decreased by feeding mugwort supplemented diets regardless of protein level and feeding period, and that of lipid increased with 12% protein diet.

• Korean Journal of Veterinary Research
• /
• v.48 no.3
• /
• pp.243-250
• /
• 2008

A calmodulin-dihydrofolate reductase (DHFR) sandwich fusion protein was generated by insertion of calmodulin into the $\beta$-bulge region of DHFR to observe the effects of structurally constraining the calmodulin structure. The calcium binding properties of the sandwich protein were almost identical to calmodulin. Similar to calmodulin ($10.7 {\mu}M$), the sandwich protein bound four equivalents of calcium, with half saturation ($K_{0.5}$) observed at a [$Ca^{2+}$] of $8{\mu}M$. However, nicotinamide adenine dinucleotide (NAD) kinase activation property of the sandwich protein was lower than that of calmodulin. The sandwich protein activated NAD kinase, but to only half of the level obtained with calmodulin. The K 0.5 for both calmodulin and the sandwich protein were approximately the same (1-2 nM). Methylation analyses of the sandwich protein show that insertion of calmodulin into DHFR results in a large decrease in methylation. The $V_{max}$ observed with the sandwich protein (95 nmole/min/ml) was only 22% of the value observed with calmodulin (436 nmol/min/ml) in the presence of calcium. Addition of trimethoprim to the reaction significantly inhibited the observed methylation rate. Overall, the data suggest that the insertion of calmodulin into the DHFR structure has little effect on calcium binding by the individual lobes of calmodulin, but may constrain the lobes in a manner that results in altered interaction with the calmodulin-dependent proteins, and severely perturbed the methyltransferase recognition site.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.37 no.1
• /
• pp.18-23
• /
• 2004

Effect of initial protein concentration on the protein removal rate was assessed for seawater aquarium using a foam separator. Protein removal rate was increased and removal efficiency was decreased with the increase of initial protein concentration. Enrichment ratio was decreased and foam generation rate was increased with the increase of initial protein concentration. Total suspended solids (TSS) removal rate was increased with the increase of initial protein concentration, and TSS removal efficiency was decreased with the increase of initial protein concentration. Turbidity removal rate and removal efficiency were increased with the increase of initial protein concentration.

• Journal of Aquaculture
• /
• v.12 no.2
• /
• pp.137-143
• /
• 1999

The effects of dietary protein level on growth, feed utlization and liver size were studied with juvenile haddock (Melanogrammus aeglefinus) of 6.88$\pm$0.54 g (mean$\pm$SD). Five isoenergetic diets containing 44.4 to 64.7% protein were fed to triplicate groups of 50 fish in a flow-through system for 6 weeks. All fish showed a relative increase of more than 300% in final weight. There were, however, no significant differences (P>0.05) in weight gain (16.1 to 17.3 g) and specific growth rate (2.9 to 3.0) among fish groups. A higher protein diets resulted in a lower feed intake. Feed: gain ratio linearly improved (P<0.05) when protein level increased from 44.4 to 59.7%. The lowest protein efficiency ratio was observed in fish fed the highest protein diet. Hepatosomatic index of fish significantly decreased from 10.7 to 7.3% with an increase in protein level. Results from this study suggest that haddock could grow well even when fed the diet with protein lower than 44.4%.

• Journal of Life Science
• /
• v.8 no.5
• /
• pp.504-508
• /
• 1998

A study was conducted to evaluate a protein removal characteristics by foam separation. The foam separator was operated in well-mixed tank which would be considered as a completely mixed condition. The feasibility of foam separation to remove protein from fresh and sea water was investigated. Protein removal characteristics of the foam separator were obtained by batch reactor operations. To find the effect of the operating parameter to protein removal rate, the foam separation was carried with variation of initial protein concentration and foam height. The results indicated that the protein removal efficiency was increased with increasing protein concentration and decreased with increasing foam height. The relationship between protein concentration and protein removal rate was evaluated by linear regression.

• Korean journal of food and cookery science
• /
• v.23 no.6
• /
• pp.810-817
• /
• 2007

The quality properties of Jeung-pyun made with soybean protein isolate(SPI) were investigated. The SPI Jeung-pyun was manufactured using 3% whole protein, 7S protein or 11S protein(w/w). The redness values (a-values) of the Jeung-pyun were negative and the yellowness(b-value) of the control group was significantly lower compared to the other samples. The textural characteristics of the Jeung-pyun were influenced by the additions of SPI. The Jeung-pyuns containing soybean flour, whole protein, 7S and 11S protein had increases in springiness, but decreases in cohesiveness. In the sensory evaluation, overall desirability, sweetness, and moistness were highest in the whole protein group. The control group showed the highest scores for hardness and toughness, and the scores for after-taste and adhesiveness were the highest in the samples made with soybean flour. The 7S and 11S additions showed high scores in terms of color and springiness. In the analyses for consumer acceptance and sensory quality intensity, the whole protein addition had significantly higher scores for general acceptability and the Makgeolli flavor. Hardness was highest for the control group and the Makgeolli flavor was strongest with the soybean flour addition.