• Journal of Microbiology and Biotechnology
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• v.30 no.12
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• pp.1810-1818
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• 2020

Inhibitor K562 (IK) protein was first isolated from the culture medium of K562 cells, a leukemia cell line, and is an inhibitory regulator of interferon-γ-induced major histocompatibility complex class II expression. Recently, exogenous truncated IK (tIK) protein showed potential as a therapeutic agent for inflammation-related diseases. In this study, we designed a novel putative anti-inflammatory peptide derived from tIK protein based on homology modeling of the human interleukin-10 (hIL-10) structure, and investigated whether the peptide exerted inhibitory effects against pro-inflammatory cytokines such as IL-17 and tumor necrosis factor-α (TNF-α). The peptide contains key residues involved in binding hIL-10 to the IL-10 receptor, and exerted strong inhibitory effects on IL-17 (43.8%) and TNF-α (50.7%). In addition, we used circular dichroism spectroscopy to confirm that the peptide is usually present in a random coil configuration in aqueous solution. In terms of toxicity, the peptide was found to be biologically safe. The mechanisms by which the short peptide derived from human tIK protein exerts inhibitory effects against IL-17 and TNF-α should be explored further. We also evaluated the feasibility of using this novel peptide in skincare products.

• Applied Biological Chemistry
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• v.28 no.3
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• pp.209-217
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• 1985

Seventeen extracellular protein producing bacteria were isolated from soil samples, among which T219 strain having a strong capability of producing the protein was selected and identified for investigation of biological characteristics. The factors which affect the protein production were investigated and the results are summarized as follows. T219 strain which produces the most extracellular protein was identified as Bacillus sp. Optimum temperature and pH for production of the extracellular protein by T219 strain were $25^{\circ}C$ and 7.5 respectively. Almost no activities of protease and amylase were observed in the protein produced by the protein producing bacteria. In the medium containing yeast extract, the cell growth was moderately high, but almost no accumulation of protein was observed. However, polypeptone had significant effects on both the cell growth and the protein accumulation. The addition of glycine and L-isoleucine to the medium containing polypeptone, yeast extract and meat extract had a great effect on the protein production; 4mg/ml of protein accumulation was observed.

• BMB Reports
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• v.38 no.6
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• pp.650-660
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• 2005

Proteins accumulated in dry, stratified Arabidopsis seeds or young seedlings, totaled 1100 to 1300 depending on the time of sampling, were analyzed by using immobilized pH gradient 2-DE gel electrophoresis. The molecular identities of 437 polypeptides, encoded by 355 independent genes, were determined by MALDI-TOF or TOF-TOF mass spectrometry. In the sum, 293 were present at all stages and 95 were accumulated during the time of radicle protrusion while another 18 appeared in later stages. Further analysis showed that 226 of the identified polypeptides could be located in different metabolic pathways. Proteins involved in carbohydrate, energy and amino acid metabolism constituted to about 1/4, and those involved in metabolism of vitamins and cofactors constituted for about 3% of the total signal intensity in gels prepared from 72 h seedlings. Enzymes related to genetic information processing increased very quickly during early imbibition and reached highest level around 30 h of germination.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.3
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• pp.186-192
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• 2004

Purpose: Human tooth proteins are highly heterogeneous, comprising diverse proteins derived from a number of genes. The attempts to identify protein for activity of tooth matrix proteins have been defied by several factors. First, the amount of proteins within teeth is very small relative to many extracellular matrix proteins of other tissues. Second, the bioassay system is tedious and needed for long time. Therefore we tried to find easy techniques, which increase the product rate, and an assay of small proteins, with which amino acid sequence is possible without additional procedures. Materials and Methods: Total protein were extracted from 300 g enamel removed teeth and 600 g teeth with 4 mol/L guanidine HCl and purified by gel chromatography. Aliquot of proteins was implanted into muscle pouches in Sprague-Dawley rats for bioassay. By SDS-PAGE and membrane blotting, molecular weight of each protein was estimated and a partial amino acid sequence was obtained. Each fraction blotted on the membrane was cut out and inserted in rat ectopic model. Results: In dissociative method, total tooth proteins were obtained 1mg/ml from enamel removed teeth and 3.5 mg/ml from teeth. In SDS-PAGE, four clear bands at the sites corresponding to 66, 40, 20 and 18 kD. Especially The 66 kD band was clearly exhibited. Amino acid sequencing from tooth could be possible using PVDF membrane blotting technique. In amino acid sequencing, 66 kD protein was identified as albumin. Conclusion: Compared with conventional method for extraction of teeth protein and bioassay of proteins, the methods in this study were easy, time-saving and more productive technique. The matured tooth proteins omitting additional procedure of mechanical removal of enamel were simply analyzed using blotted PVDF membrane. This method seems to make a contribution as a technique for bioassay and amino acid sequencing of protein.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.337-345
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• 2014

Pichia pastoris is one of the most widely used expression systems for the secretory expression of recombinant proteins. The secretory expression in P. pastoris usually makes use of the prepro $MAT{\alpha}$ sequence from Saccharomyces cerevisiae, which has a dibasic amino acid cleavage site at the end of the signal sequence. This is efficiently processed by Kex2 protease, resulting in the secretion of high levels of proteins to the medium. However, the proteins that are having the internal accessible dibasic amino acids such as KR and RR in the coding region cannot be expressed using this signal sequence, as the protein will be fragmented. We have identified a new signal sequence of 18 amino acids from a P. pastoris protein that can secrete proteins to the medium efficiently. The PMT1-gene-inactivated P. pastoris strain secretes a ~30 kDa protein into the extracellular medium. We have identified this protein by determining its N-terminal amino acid sequence. The protein secreted has four DDDK concatameric internal repeats. This protein was not secreted in the wild-type P. pastoris under normal culture conditions. We show that the 18-amino-acid signal peptide at the N-terminal of this protein is useful for secretion of heterologous proteins in Pichia.

• Nutrition Research and Practice
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• v.3 no.3
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• pp.200-207
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• 2009

This study examined the effects of com gluten (CG) and its hydrolysate consumptions on weight reduction in rats fed a high-fat diet. Eight-month-old male Sprague-Dawley rats (n=40) were fed a high-fat diet (40% calorie as fat) for 4 weeks. They were then randomly divided into four groups and fed the isocaloric diets with different protein sources for 8 weeks. The protein sources were casein (control group), intact CG (CG group), CG hydrolysate A (CGHA group, 30% of protein as peptides and 70% as free amino acids) and CG hydrolysate P (CGHP group, 93% of protein as peptides and 7% as free amino acids). Body weight gain, adipose tissue weights, nitrogen balance, absorptions of energy, protein and fat, lipid profiles in plasma, liver and feces and hepatic activities of camitine palmitoyl transferase (CPT), fatty acid synthase (FAS), malic enzyme (ME) and glucose-6-phosphate dehydrogenase (G6PDH) were assessed. The CGHA diet had the highest amount of BCAAs, especially leucine, and most of them existed as free amino acid forms. The CGHA group showed significant weight reduction and negative nitrogen balance. Protein absorption and apparent protein digestibility in the CGHA group were significantly lower than those in other groups. Adipose tissue weights were the lowest in the CGHA group. Activity of CPT tended to be higher in the CGHA group than in other groups and those of FAS, ME and G6PDH were significantly lower in the CGHA group than in other groups. In conclusion, the CGHA diet which had relatively high amounts of free amino acids and BCAAs, especially leucine, had a weight reduction effect by lowering adipose tissue weight and the activities of FAS, ME and G6PDH in experimental animals, but it seemed to be a negative result induced by lowering protein absorption, increasing urinary nitrogen excretion and protein catabolism.

• Journal of The Korean Society of Grassland and Forage Science
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• v.29 no.2
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• pp.129-136
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• 2009

This study was to optimize the harvest time of several winter cereal forage crops. Barley, rye, oat, triticale and wheat were evaluated for the quantity and quality of hay and silage at four different harvesting stages, including heading, 10 days after heading (DAH), 20 DAH and 30 DAH. Barley and wheat harvested at 20 DAH, and oat and triticale at 30 DAH showed maximum dry matter yield. Crude protein content of barley, wheat, triticale and oat decreased significantly with advancing crop maturity. Mean crude protein content was the highest in rye and the lowest in oat. The NDF and ADF content of barley, wheat, triticale and oat decreased with late harvest, while rye increased. Maximum total digestible nutrients (TDN) content was recorded in barley and wheat harvested at 20 DAH, in triticale and oat at 30 DAH, and in rye at heading. The maximum TDN content of silage, 66.98%, was recorded in barley, followed by wheat, triticale, oat and rye. However, The crude protein, NDF and ADF of silage was 1 to 4 times higher than those of hay. In this study, barley harvested at 20-25 DAH, wheat and triticale at 30 DAH, and rye at heading provided a good compromise between dry matter yield and forage quality. At this stage, a sufficient quantity of fodder with moderate forage quality was obtained.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.117-121
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• 2004

The patterns of diapause-associated proteins of silkworm eggs were analyzed by two-dimensional (2-D) gel electrophoresis. Among the hundreds of spots on the 2-D gels, at least two proteins were considered to be associated with diapause. A protein, spot 4, with an approximate molecular weight of 38 kDa and pI 6.1 was observed in the HCI-treated, cold-treated, and diapause eggs, respectively. Spot 4 was undetectable in unfertilized eggs and non-diapause eggs at two days after oviposition, suggesting that this protein may be associated with the entrance to diapause. A protein, spot 11, with an approximate molecular weight of 21 kDa and pI of 61 was detected in the unfertilized, HCl-treated, and cold-treated eggs, respectively, after oviposition by normal moths. In diapausing eggs, a protein corresponding to spot 11 was observed in 3-, 5-, and 30-day-old eggs, while the protein was not detected one-day-old eggs. The protein corresponding to spot 11 was not detected in unfertilized and non-diapause eggs obtained from subesophabeal ganglion (SG)-extirpated moths either. Spot 11 was also considered to be a diapause specific protein, which occurred at only early embryonic stage under the control of diapause-downregulated gene.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.287-299
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• 1994

Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Subpopulations expressing major histocompatibility complex(MHC) class I antigen were $96.2{\pm}3.1%$ and $86.6{\pm}3.8%$ in piglets fed with plasma protein and in piglets fed without plasma protein, respectively. 2. Proportion of leukocyte subpopulation expressing MHC class II antigens were significantly higher in the piglets fed with plasma protein than ones without plasma protein. The proportion was $27.6{\pm}3.6%$ and $16.6{\pm}2.2%$ in MHC class II DQ antigen, and $28.1{\pm}2.0%$ and $20.0{\pm}0.3%$ in MHC class II DR antigen, respectively. 3. A significant increase in the proportion of cells expressing poCD2 was not found in piglets fed plasma protein. 4. Proportion of subpopulation expressed porcine(Po) CD4 antigens which specific to helper T lymphocytes were not increased (18.3-19.1% vs. 25.6-28.8%), rather slightly decreased, in plasma protein-treated group. 5. The most important increase of proportion in plasma protein-treated group was the leukocyte subpopulation specific to $poCD8^+$ T cytotoxic/suppressor lymphocytes. The expression level was significantly higher up to 45.9-47.1% in plasma protein-treated group in comparing with 29.7-33.0% in non-plasma protein-treated group. 6. Lymphoblastogenetic responses using different concentrations of Con A mitogen and plasma protein has found that the responses of lymphocyte from piglets fed plasma protein was significantly activated (p<0.01). The activities measured by 3[H]-thymidine incorporation showed 3-6 times stronger in plasma protein-treated group than those in non-plasma protein-treated group. The study has concluded that plasma protein, which has known as a nonspecific immunostimulator, may have an immunoenhancing activities in porcine lymphoid system by increase the activated cell proportions and their blastogenetic properties which is critical to host immune responses.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.1
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• pp.30-35
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• 2014

This study was conducted to evaluate the protein requirement for maintenance of 2-year-old female Korean spotted deer. In the course of the experiment, each of three hand-reared female spotted deer was fed three diets that were iso-calorically formulated to contain low (approximately 7%), medium (12%), and high (17%) levels of crude protein (CP). Each of six trials included a 5-day transition, a 10-day preliminary, and a 7-day collection period. Dietary protein levels affected the apparent digestibility of CP (p<0.05) but not the apparent digestibility of dry matter, organic matter, or acid detergent fiber. All of the deer showed a positive CP balance on all of the diets. The maintenance CP requirement estimated by regression analysis was 4.17 g/kg metabolic body weight $(W^{0.75}){\cdot}d$. The maintenance digestible CP requirement was 1.42 g/kg $W^{0.75}{\cdot}d$. The metabolic fecal CP was 1.95 g/kg $W^{0.75}{\cdot}d$. The blood urea nitrogen of spotted deer increased (p<0.05) as the dietary protein levels increased.