Fatty acid composition and electrophoretic patterns of protein of Korean soybeans were studied, The protein content showed an inverse relation with the lipid content, The content of lipid types of total lipid was in the order of neutral lipid, glycolipid and phospholipid, Cultivar 102-B had higher content of neutral lipid but lower content of phospholipid compared with others. The main component of neutral lipid was triglyceride followed by sterol. The main fatty acids of total lipid were linoleic, oleic and palmitic acids, which comprised over 85% of total fatty acids. The main fatty acid of neutral lipid, glycolipid and phospholipid were linoleic acid. The polyacrylamide gel electrophoretic pattern of protein showed seven bands. The cultivar 102-B had the highest content of IIS globulin. Sodium-dodecyl sulfate PAG electrophoresis demonstrated that all soybeans had similar composition of subunit.
This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components in the MDBK(Mardin-Darby Bovine Kidney) cells. For the In vitro test, MDBK cells in ${\alpha}$-MEM(Minimum Essential Medium) were divided into 4 treatment groups according to drug types and dosages as follows; the control(group A), group B was treated with 0.3mM caffeine, group C was treated with 0.3mM caffeine and 0.3mM ferric chloride, group D was treated with 0.3mM caffeine and 0.3mM vitamin E. Those groups were further divided into 5 subgroups according to the time lapsed(control, 4hrs, 8hrs, 24hrs and 48hrs lapsed group). The concentrations of the carbonyl group and malondialdehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) and fatty acid compositions were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the MDBK cells. The results obtained from this study were summarized as follows; 1. The concentrations of carbonyl group and malondialdehyde in MDBK cells of group C were significantly higher(p<0.01) in comparison to the control, and increased according to the time lapsed. But the results of groups B and D were little different in comparison to the group C. 2. As the analytical results of fatty acid compositions in MDBK cells, the proportions of palmitoleic acid and linoleic acid in groups B, C and D were lower in comparison to the control, while the proportion of arachidonic acid in groups B, C and D were significantly higher(p<0.01) in comparison to the control. 3. In order to determine the oxidative damages to the protein in MDBK cells, the patterns of the SDS-PAGE were examined and the patterns of SDS-PAGE in groups C and D were significantly different between 43kd and 200kd of molecular weight.
Li, Xiaoguang;Zhang, Qian;Gan, Longzhan;Jiang, Guangyang;Tian, Yongqiang;Shi, Bi
Journal of Microbiology and Biotechnology
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v.32
no.1
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pp.99-109
/
2022
This study is the first report on production and characterization of the enzyme from an Ornithinibacillus species. A 4.2-fold increase in the extracellular protease (called L9T) production from Ornithinibacillus caprae L9T was achieved through the one-factor-at-a-time approach and response surface methodological optimization. L9T protease exhibited a unique protein band with a mass of 25.9 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This novel protease was active over a range of pH (4-13), temperatures (30-80℃) and salt concentrations (0-220 g/l), with the maximal activity observed at pH 7, 70℃ and 20 g/l NaCl. Proteolytic activity was upgraded in the presence of Ag+, Ca2+ and Sr2+, but was totally suppressed by 5 mM phenylmethylsulfonyl fluoride, which suggests that this enzyme belongs to the serine protease family. L9T protease was resistant to certain common organic solvents and surfactants; particularly, 5 mM Tween 20 and Tween 80 improved the activity by 63 and 15%, respectively. More importantly, L9T protease was found to be effective in dehairing of goatskins, cowhides and rabbit-skins without damaging the collagen fibers. These properties confirm the feasibility of L9T protease in industrial applications, especially in leather processing.
Ku, Su-Kyung;Kim, Jake;Kim, Se-Myung;Yong, Hae In;Kim, Bum-Keun;Choi, Yun-Sang
Food Science of Animal Resources
/
v.42
no.3
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pp.441-454
/
2022
This study aimed to determine the effect of enzyme, guar gum, and pressure processing on the digestibility and physicochemical properties of age-friendly liver sausages. Liver sausages were manufactured by adding proteolytic enzyme (Bromelain) and guar gum, and pressure-cooking (0.06 MPa), with the following treatments: control, without proteolytic enzyme; T1, proteolytic enzyme; T2, proteolytic enzyme and guar gum; T3, pressure-cooking; T4, proteolytic enzyme and pressure-cooking; T5, proteolytic enzyme, guar gum, and pressure-cooking. The pH was high in the enzyme- and pressure-processed groups. The pressure-processed groups had lower apparent viscosity than other cooking groups, and it decreased during enzyme treatment. Hardness was lower in the enzyme- and pressure-processed groups than in the control, and the T4 was the lowest. Digestibility was the highest in T4 at 82.58%, and there was no significant difference with that in T5. The general cooking group with enzyme and guar gum also showed higher digestibility than the control (77.50%). As a result of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme- and pressure-treated groups (T4, T5) were degraded more into low-molecular-weight peptides (≤37 kDa) than the control and other treatments. Viscoelasticity showed similar trends for viscous and elastic moduli. Similarly, combined pressure processing and enzymatic treatment decreased viscoelasticity, while guar gum increased elasticity but decreased viscosity. Therefore, the tenderized physical properties and improved digestibility by enzyme and pressurization treatment could be used to produce age-friendly spreadable liver sausages.
Shine Htet Aung;Edirisinghe Dewage Nalaka Sandun Abeyrathne;Mahabbat Ali;Dong Uk Ahn;Young-Sun Choi;Ki-Chang Nam
Food Science of Animal Resources
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v.43
no.1
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pp.46-60
/
2023
Slaughterhouse blood is a by-product of animal slaughter that can be a good source of animal protein. This research purposed to examine the functional qualities of the blood plasma from Hanwoo cattle, black goat, and their hydrolysates. Part of the plasma was hydrolyzed with proteolytic enzymes (Bacillus protease, papain, thermolysin, elastase, and α-chymotrypsin) to yield bioactive peptides under optimum conditions. The levels of hydrolysates were evaluated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis. The antioxidant, metal-chelating, and angiotensin I-converting enzyme (ACE) inhibitory properties of intact blood plasma and selected hydrolysates were investigated. Accordingly, two plasma hydrolysates by protease (pH 6.5/55℃/3 h) and thermolysin (pH 7.5/37℃/3-6 h) were selected for analysis of their functional properties. In the oil model system, only goat blood plasma had lower levels of thiobarbituric acid reactive substances than the control. The diphenyl picrylhydrazyl radical scavenging activity was higher in cattle and goat plasma than in proteolytic hydrolysates. Ironchelating activities increased after proteolytic degradation except for protease-treated cattle blood. Copper-chelating activity was excellent in all test samples except for the original bovine plasma. As for ACE inhibition, only non-hydrolyzed goat plasma and its hydrolysates by thermolysin showed ACE inhibitory activity (9.86±5.03% and 21.77±3.74%). In conclusion, goat plasma without hydrolyzation and its hydrolysates can be a good source of bioactive compounds with functional characteristics, whereas cattle plasma has a relatively low value. Further studies on the molecular structure of these compounds are needed with more suitable enzyme combinations.
Seonmin Lee;Kyung Jo;Hyun Gyung Jeong;Seul-Ki-Chan Jeong;Jung In Park;Hae In Yong;Yun-Sang Choi;Samooel Jung
Food Science of Animal Resources
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v.43
no.2
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pp.305-318
/
2023
This study investigated the protein digestibility of chicken breast and thigh in an in vitro digestion model to determine the better protein sources for the elderly in terms of bioavailability. For this purpose, the biochemical traits of raw muscles and the structural properties of myofibrillar proteins were monitored. The thigh had higher pH, 10% trichloroacetic acid-soluble α-amino groups, and protein carbonyl content than the breast (p<0.05). In the proximate composition, the thigh had higher crude fat and lower crude protein content than the breast (p<0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of myofibrillar proteins showed noticeable differences in the band intensities of tropomyosin α-chain and myosin light chain-3 between the thigh and breast. The intrinsic tryptophan fluorescence intensity of myosin was lower in the thigh than in the breast (p<0.05). Moreover, circular dichroism spectroscopy of myosin revealed that the thigh had higher α-helical and lower β-sheet structures than the breast (p<0.05). The cooked muscles were then chopped and digested in the elderly digestion model. The thigh had more α-amino groups than the breast after both gastric and gastrointestinal digestion (p<0.05). SDS-PAGE analysis of the gastric digesta showed that more bands remained in the digesta of the breast than that of the thigh. The content of proteins less than 3 kDa in the gastrointestinal digesta was also higher in the thigh than in the breast (p<0.05). These results reveal that chicken thigh with higher in vitro protein digestibility is a more appropriate protein source for the elderly than chicken breast.
Jae Won Jeong;Seung Yun Lee;Da Young Lee;Jae Hyeon Kim;Seung Hyeon Yun;Juhyun Lee;Ermie Jr. Mariano;Sung Sil Moon;Sun Jin Hur
Food Science of Animal Resources
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v.44
no.3
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pp.533-550
/
2024
Peptides with bioactive effects are being researched for various purposes. However, there is a lack of overall research on pork-derived peptides. In this study, we reviewed the process of obtaining bioactive peptides, available analytical methods, and the study of bioactive peptides derived from pork. Pepsin and trypsin, two representative protein digestive enzymes in the body, are hydrolyzed by other cofactors to produce peptides. Bicinchoninic acid assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, chromatography, and in vitro digestion simulation systems are utilized to analyze bioactive peptides for protein digestibility and molecular weight distribution. Pork-derived peptides mainly exhibit antioxidant and antihypertensive activities. The antioxidant activity of bioactive peptides increases the accessibility of amino acid residues by disrupting the three-dimensional structure of proteins, affecting free radical scavenging, reactive oxygen species inactivation, and metal ion chelating. In addition, the antihypertensive activity decreases angiotensin II production by inhibiting angiotensin converting enzyme and suppresses blood pressure by blocking the AT1 receptor. Pork-derived bioactive peptides, primarily obtained using papain and pepsin, exhibit significant antioxidant and antihypertensive activities, with most having low molecular weights below 1 kDa. This study may aid in the future development of bioactive peptides and serve as a valuable reference for pork-derived peptides.
Objective: Although pork loins is not a tough meat, they need to develop meat products with a soft texture for the elderly. This study focused on the physicochemical properties and tenderness characteristics of pork loin injected with green kiwifruit juice (GRJ) and gold kiwifruit juice (GOJ) during various incubation times. In addition, the antioxidant activities of hydrolysate derived from the hydrolysis of pork loin by kiwifruit juice protease were evaluated. Methods: The pork loin was injected with 10% and 20% GRJ and GOJ, under various incubation times (0, 4, 8, and 24 h). Then, the physicochemical properties and tenderness of pork loins were measured. 2,2- diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power were conducted to determine hydrolysate's antioxidant activities derived from pork loin's hydrolysis by kiwifruit juice protease. Results: GRJ had greater tenderizing ability than GOJ, even at the 10% addition. When kiwifruit juice was injected into pork loin, the tenderness increased with increasing incubation time. This was confirmed by the decrease in intensity of the myosin heavy chain (MHC) band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In particular, the MHC band decreased at 8 h for both 10% GRJ and 20% GOJ and at 4 h for 20% GRJ alone. The highest myofibril fragmentation index and peptide solubility were observed in pork loin treated with 20% GRJ compared to the other treatments during incubation. The 10% GRJ and 20% GOJ treatments showed similar levels of antioxidant activity of the protein hydrolysates in pork loin, and 20% GRJ showed the highest activity among the treatments. Conclusion: Kiwifruit juice had protease activity, and GRJ was more useful for tenderizing meat products than GOJ. Thus, GRJ at 10% could be a potential agent to tenderize and enrich the natural antioxidant activity through the proteolysis of pork loin.
Purpose: Due to the many problems with commercially available vaccines, the production of effective vaccines against brucellosis is a necessity. The aim of this study was to evaluate the immune responses caused by the chimeric protein consisting of trigger factor, Bp26, and Omp31 (TBO) along with aluminum hydroxide (AH/TBO) and selenium (Se/TBO) nanoparticles (NPs) as adjuvants in mouse model. Materials and Methods: Recombinant antigen expression was induced in Escherichia coli BL21 (DE3) bacteria using IPTG (isopropyl-d-1-thiogalactopyranoside). Purification and characterization of recombinant protein was conducted through NiFe3O4 NPs, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot. NP characteristics, including morphology and particle size, were measured in vitro. The recombinant TBO was loaded on to AH and Se NPs and were administered subcutaneously. After mice immunization, measurement of antibody titter and protection assay was performed. Results: The average sizes of AH and Se NPs were about 60 nm and 150 nm, respectively. The enzyme-linked immunosorbent assay results showed that the serum of mice immunized by subcutaneous injection with both nanovaccines produced significant immunoglobulin G (IgG) responses against the chimeric antigen. The results of TBO-specific IgG isotype (IgG2a/IgG1) analysis showed that both AH and Se NPs induced a type to T-helper immune response. In addition, the results of the challenge with the pathogenic strain of Brucella melitensis 16M showed that vaccinated mice with AH/TBO NPs indicated a higher reduction of bacterial culture than immunized mice with Se/TBO NPs and TBO alone. Conclusion: The results showed that AH NPs carrying chimeric antigen can be a promising vaccine candidate against brucellosis by producing protective immunity.
Oh, Ryunkyoung;Moon, Soo Young;Cho, Hwa Jin;Jang, Won Je;Kim, Jang-Ho;Lee, Jong Min;Kong, In-Soo
Microbiology and Biotechnology Letters
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v.44
no.3
/
pp.355-362
/
2016
The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose-1-phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.
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