• Microbiology and Biotechnology Letters
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• v.50 no.4
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• pp.508-511
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• 2022

In this study, Phaseolus vulgaris (kidney bean) leghemoglobin a (PhLba) gene was cloned into pPICZαA and expressed in Pichia pastoris to sustainably produce a heme-carrying protein for organoleptic use in plant-based meat. The recombinant PhLba protein was secreted into the culture medium in a solubilized form, and the molecular weight of the purified PhLba was estimated to be 16.5 kDa using SDS-PAGE. In addtion, the yield of recombinant PhLba holoprotein was enhanced by supplementation of the cultivation medium with hemin. This result indicates that the apo-forms of PhLba can be effectively saturated with cofactor.

• Childhood Kidney Diseases
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• v.13 no.2
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• pp.170-175
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• 2009

Purpose : FSGS do not respond well to any kind of therapy and gradually progress to end-stage renal disease. This study was conducted to investigate the difference of protein expression between MCNS and FSGS as a preliminary study for understanding the pathophysiology of FSGS. Methods : Renal biopsy samples of MCNS and FSGS were obtained, which was diagnosed by one pathologist. They were solubilized with a conventional extraction buffer for protein extraction. The solution was applied on immobilized linear gradient strip gel (pH 4-7) using IPGphor system. Silver staining was carried out according to standard method. Protein identification was done by searching NCBI database using MASCOT Peptide Mass Fingerprint software. Results : The differences in protein expressions between MCNS and FSGS were shown by increased or decreased protein spots. Most prominently expressed spot among several spots in FSGS was isolated and analyzed, one of which was glutathione S-transferase (GST) P1-1, whereas it was not found in MCNS. So GSTP1-1 was considered as the one of the key biomarkers in pathogenesis of FSGS. Conclusion : This result would be helpful in diagnosing FSGS and researching FSGS. Further studies for glutathione S-transferase P1-1 might be necessary to elucidate the mechanisms regarding FSGS.

• Korean Journal of Food Science and Technology
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• v.21 no.4
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• pp.542-548
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• 1989

For the selective separation of proteins, the solubilization and desolubilization of proteins in sodium-di-2-ethylhexyl sulfosuccinate (AOT)-isooctane reverse micellar system were investigated. Protein solubilization increased with increasing the concentration of AOT to 200 mM and then decreased above that concentration. Protein was solubilized into reverse micelles in the pH range below the isoelectric Point of each protein, pH 4-10 for lysozyme and pH 5-6 for trypsin and ${\alpha}-chymotrypsin$, Lysozyme, trypsin and ${\alpha}-chymotrypsin$ were efficiently extracted in the precence of KCl and NaCl while larger molecular weight proteins such as pepsin and BSA had high solubilization with $CaCl_2$. At higher ionic strength all proteins exhibited murk less tendency to solubilize and the increase of ionic strength resulted in the decrease of micelle size. Lysozyme was successfully back transfered at pH 12.2 and 1.0M KCl; trypsin at pH 12.6 and 0.5M KCl; and ${\alpha}-chymotrypsin$ at pH 6.7 and 0.5M KCl. In a test group separation experiments, complete separation of lysozyme from BSA could be obtained.

• Korean Journal of Food Science and Technology
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• v.19 no.6
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• pp.499-505
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• 1987

The functional properties of lupin seed protein concentrate (LPC) were examined and compared to those of soybean protein isolate (SPI) and Na-caseinate. LPC-50, of which protein level was 50%, was prepared by a two phase solvent (hexane: alcohol: water= 10:7:3) extraction method. LPC-70 was made from LPC-50 by removing the fractions solubilized by carbohydrate decomposing enzymes. The solubilities of LPC-50 and LPC-70 were similar to that of of SPI but slightly higher at pH 4-5, and less susceptible to the added salt. The apparent viscosity of LPC increased exponentially as the concentration increased over 6% level, and the change was similar to that of Na-caseinate. LPC showed strong pseudoplastic non-Newtonean flow behavior, which was similar to that of SPI The emulsifying capasity of LPC-70 was similar to that of SPI when salt was added. The foaming capacity of LPC was comparable to that of SPI. LPC showed high oil and water absorption capacities, which increased as the protein level was elevated. LPC-70 showed the highest oil absorption capacity of all the samples tested.

• Journal of Sericultural and Entomological Science
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• v.31 no.1
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• pp.37-44
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• 1989

The parasporal crystals of Bacillus thuringinsis subspecies kurstaki, dendrolimus, finitimus, aizawai and israelensis were compared by polyacrylamide electrophoresis, amino acid composition and immunological analysis. In the subspecies of kurstaki, dendrolimus, finitimus and aizawai, the molecular weights of main subusnits of crystal solubilized by alkaline solution were 1.3${\times}$105 and 6.5${\times}$104 while those of subsp. israelensis were 4${\times}$104 and 1,4${\times}$104. The degradation of lepidopteran toxic subspecies crystals by silkworm midgut protease showed 6.0-6.4${\times}$104 molecular weight and the pattern of degradation of subsp. israelensis crystals was similar to that of alkaline solution treatment. In the amino acid composition, aspartic acid in subsp. israelensis and glutiamic acid in the other four subspecies were the most abundant. The immunological characteristics of the crystals revealed that the antibody produced against the alkali-solubilized crystal protein of subsp. israelensis reacted with only its antigen, but the crystal antigens from the other four lepidopteran toxic subspecies did cross-react with each other as well as with their own homologous antisera.

• IMMUNE NETWORK
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• v.1 no.3
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• pp.250-259
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• 2001

To determine if initial infection with Mycobacterium tuberculosis changes the balance of cytokines between T cells and macrophages, we evaluated interferon (IFN)-${\gamma}$), interleukin-12 (IL)-12, and tumor necrosis factor (TNF)-${\alpha}$ productions by peripheral blood mononuclear cells (PBMC) from 15 untreated active pulmonary tuberculosis (TB) patients and 12 healthy tuberculin reactors (HTR). Freshly isolated PBMC were stimulated with Triton X-100 solubilized protein (TSP), 30-kDa or purified protein derivatives (PPD) antigen for 6, 18 and 96 hours. IL-12 p40 production by antigen-stimulated PBMC from TB patients was significantly decreased compared with that in HTR. In addition, IFN-${\gamma}$ production was significantly depressed in TB patients than that in HTR at a 96-hr stimulation. However, TNF-${\alpha}$ production was significantly higher in antigen-stimulated PBMC from TB than that of HTR. A pronounced increase in IFN-${\gamma}$ protein followed neutralization of IL-10 in early TB patients. However, neutralization of TNF-${\alpha}$ did not significantly alter IFN-${\gamma}$ induction in PBMC from TB patients. There were no significantly differences in the cytokine productions among three proteins, TSP, 30-kDa or PPD antigen. These results indicate that development of TB may be strongly associated with dysregulated productions of IL-12, IFN-${\gamma}$ and TNF-${\alpha}$, during the initial immune responses to M. tuberculosis. Further understanding of operative cytokine networks during human immune cell responses to protein antigens of M. tuberculosis may improve strategies for vaccine development.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.702-708
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• 1998

The effects of mushrooms such as Lentinus edodes and Pleurotus ostreatus on anti-cancer activity through in vivo and in vitro experiments were powders of protein-bound polysaccharides in mushrooms were solubilized in 0, 5, 25 mg/kg saline, respectively and were used in vitro experiments. The in vivo experiments were carried out as followed: i) anti-cancer activities on Leukemia $(L_{1210})$, Hepatlicus cancer $(H_{22})$ and Sarcoma180/(S180), and ii) the effect on immune system through changes in intestine weight and the number of hemolytic plague forming cells. Protein-bound polysaccharides of all showed anti-cancer activity on $L_{1210}$ and fruit body of Lentinus edodes 25 mg/kg treatment group showed the highest inhibition rate (86%). Pleurotus ostreatus mycelial in medium of cultivate 25 mg/kg treatment. Fruid body of Lentinus edodes 25 mg/kg treatment group showed the highest inhibition rate (86% and 71%, respectively) on $H_{22}$ among them. The inhibition rates of fruit body and mycelial of Lentinus edodes 25 mg/kg treatment groups on S180 were 33.9% and 30.9%, respectively. Each samples of 50, 100, 200, $400\;{\mu}g/{\mu}L$ on in vitro cell toxicity test did not show significantly different cell death rates at P<0.05. In immune test, weights of liver and spleen were increased according to increase in conc. but were not significantly different at P<0.05. The weights of thymus were heavy in fruit body and mycelial of Lentinus edodes treatment group but were not significantly different at P<0.05. Hemolytic plague forming cells with antibody formation capability were significantly high in fruit body and mycelial of Lentinus edodes treatment samples.

• Journal of Plant Biotechnology
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• v.48 no.3
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• pp.165-172
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• 2021

The solubilization of isolated proteins into the adequate buffer containing of surfactants is primary step for proteomic analysis. Particularly, sodium dodecyl sulfate (SDS) is the most widely used surfactant, however, it is not compatible with mass spectrometry (MS). Therefore, it must be removed prior to MS analysis through rigorous washing, which eventually results in inevitable protein loss. Recently, photocleavable surfactant, 4-hexylphenylazosulfonate (Azo), was reported which can be easily degraded by UV irradiation and is compatible with MS during proteomic approach using animal tissues. In this study, we employed comparative label-free proteomic analysis for evaluating the solubilization efficacies of the Azo and SDS surfactants using rice leave proteins. This approach led to identification of 3,365 proteins of which 682 proteins were determined as significantly modulated. Further, according to the subcellular localization prediction in SDS and Azo, proteins localized in the chloroplast were the major organelle accounting for 64% of the total organelle in the SDS sample, while only 37.5% of organelle proteins solubilized in the Azo were predicted to be localized in chloroplast. Taken together, this study validates the efficient solubilization of total protein isolated from plant material for bottom-up proteomics. Azo surfactant is suitable as substitute of SDS and promising for bottom-up proteomics as it facilitates robust protein extraction, rapid washing step during enzymatic digestion, and MS analysis.

• Journal of Sericultural and Entomological Science
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• v.34 no.1
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• pp.41-48
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• 1992

Dissolution and degradation of the parasporal crystal proteins produced from B. thuringiensis var. kurstaki, B. thuringiensis var. dendrolimus and B. thuringiensis var. aizawai were investigated. SDS-polyacrylamide gel electrophoresis analysis showed that the crystals contained major protein with molecular weight of approximately 134 kDa for B. thuringiensis var. aizawai 143 kDa for B. thuringiensis var. kurstaki and 149 kDa for B. thuringiensis var. dendrolimus, respectively. Crystals of three other strains were incubated alkali solutions at various pH or gut juice of Silkwarm Bombyx mori, Fall webworm Hyphantria cunea, and Common Cabbage worm Pieris rapae. When crystals of these strains were solubilized by alkali solutions, no major differences among strains B. thuringiensis could be detected. Among the strains studied, crystal protins (130-66 kDa) consist of protease resistant polypeptides in the 45-66 kDa size range when treated with gut juice of three insect species.

• KSBB Journal
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• v.24 no.6
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• pp.563-569
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• 2009

Fibroin is an insoluble structural protein from Bombyx mori. It can be solubilized by dissolving in a hot $CaCl_2$ solution and subsequent dialysis. The aqueous solution is unstable and a transition from aqueous fibroin molecules rich in random coil is undergo to one rich in $\beta$-sheet content, resulting in hydrogelation. However, fibroin gel is so fragile and plastic that its mechanical property should be reformed for various applications. In this report, a semi-solid form of fibroin gel was prepared using glycerol and ethanol and was investigated to analyze their flow properties. A fibroin gel with 80% glycerol showed pseudoplastic and thixotropic properties. The square root of its yield stress varied linearly with fibroin concentration and it extrapolated to zero shear stress at 0.2% fibroin. A fibroin gel with 40% ethanol was shown to be highly thixotropic but its shear-thinning behavior was only observed above a certain level of shear rate. Its pseudoplasticity was restored by a high rate of shear stress.