• Preventive Nutrition and Food Science
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• v.3 no.4
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• pp.379-381
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• 1998

An angiotensin converting enzyme (ACE) inhibitor was isolated and partially purified from bovine blood plasma. Bovine blood plasma was obtained after removing blood cells by centrifugation, followed by the addition of anticoagulant to whole bovine blood. To precipitate plasma proteins, bovine blood plasma was treated with 4% trichloroacetic acid (TCA) as a final concentration .An ACE inhibitor was isolated from TCA supernatnat, using ultrafiltration, gel permeation chormatography, and reverse-phase high pressure liquid chromatogrpahy. The ACE inhibitor purified from TCA supernatant had IC50 values of 9.4$\mu$M.

• Applied Biological Chemistry
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• v.40 no.1
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• pp.39-42
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• 1997

Angiotensin converting enzyme (ACE) inhibitors were isolated and purified from pig blood plasma. Pig blood plasma was obtained after removing blood cells by centrfugation, followed by the addition of anticoagulant to whole pig blood. To precipitate plasma proteins, pig blood plasma was treated with 4% trichloroacetic acid (TCA) as a final concentration. ACE inhibitors were isolated from plasma protein hydrolysates and TCA supernatant, using ultrafiltration, gel permeation chromatography, and reverse-phase high pressure liquid chromatography. ACE inhibitors isolated from plasma hydrolysates and TCA supernatant had $IC_{50}$ values of $23\;{\mu}M$ and $2\;{\mu}M$, pentapeptide, respectively.

• Applied Biological Chemistry
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• v.39 no.2
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• pp.123-126
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• 1996

Simple and rapid method of purification of plasma proteins from bloods of slaughtered animals was developed and the proteins were applied to plywood products as a blood 히ue to utilize waste materials. Plasma protein was obtained by adding 2% trichloroacetic acid (TCA) or 0.6 N HCI as optimal concentration to the supernatant, after centrifugation of bloods. Molecular properties of beef and pig plasma proteins were examined on SDS-PAGE. Application of blood glue to plywood was quite satisfactory compared to the synthetic amino resin by tensile-shear test for the strength of adhesive bonding.

• KSBB Journal
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• v.18 no.5
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• pp.371-376
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• 2003

Antioxidative and cholesterol-reducing activity of Bacillus polyfermenticus SCD were measured to characterize its probiotic properties. DPPH (1,1-diphenyl-2-picyryl hydrazyl) radical scavenging activity of the culture supernatant of B. polyfermenticus SCD was estimated to be 48%. The culture supernatant on the peroxidation of linoleic acid were investigated and the value was shown to be about 45%. The inhibition of TBA (2-thiobarbituric acid) formation of the culture supernatant was revealed 60% when stimulator was presented. The SOD-like activity of the culture supernatant was about 15%, which is similar to BHT (butylated hydroxytoluene) and ${\alpha}$-tocopherol. After cultured in TSB broth added soluble cholesterol either 0.1% or 0.3% of oxgall in 37$^{\circ}C$ for 24 h aerobically, cholesterol-reducing activities were revealed about 67% or 64%, respectively. To test whether the products are cholesterol-related or not, residual activity was determined. The cholesterol activity was rarely changed. In addition, when the cell extracts recovered after cultivation, was tested in absence of cholesterol, cholesterol activity was not detected. However, cholesterol activity was detected in the presence of cholesterol. Thus, it was assumed that B. polyfermenticus SCD could reduce cholesterol by conjugating with it, rather than by digesting the cholesterol using cholesterol-hydrolyzing enzymes.

• KSBB Journal
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• v.18 no.2
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• pp.94-99
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• 2003

The effect of medium composition on the production of bacterial cellulose (BC) by Gluconacetobacter hansenii PJK was investigated. The addition of yeast extract and peptone in the medium increased the production yield (Y/sub p/s/) of BC. The amount of BC produced by G. hansenii PJK was constant if the initial pH of the medium was in the range 4.5 to 6.0. Strains from the supernatant of the culture medium produced more BC than those from inside the BC. BC production was dependent on glucose metabolism, and the addition of fructose or lactate as a carbon source converted cells to Cel/sup -/ mutants. Cel/sup -/ mutants produced by the addition of fructose or lactate to the medium caused 73% or 30% decreases in BC production, respectively. The addition of succinate, which is one of the constituents of the TCA cycle, did not affect the production of BC.

• Journal of the Korean Society of Food Science and Nutrition
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• v.10 no.1
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• pp.85-91
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• 1981

The growing cells of S. aureus were fractionated along the Schmidt-Thannhauser-Schneider's technique into several fractions such as TCA(trichloroacetic acid)-soluble, lipid, nucleic acid, protein and residue fraction. They were also fractionated according to their cellular structure into Sonic-supernatant, SDS(sodium lauryl sulfate)-soluble, Formamide-soluble and Residue fraction. Fractionation was carried out by orderly treatment of the Sonic pellet with 1.0% SDS and hot$(150^{\circ}C)$ formamide, and the pellet was prepared by centrifugation of the cells sonic osillated for 20 minutes at 150 watt. Sonic-supernatant fraction contained a 91.3% of total DNA while other fractions contained less than 9.5%. SDS-soluble fraction showed a high activity of malate dehydrogenase(13.67 unit/mg protein) and which was higher 22.3 times than the activity found from unsoluble fraction. Formamide-soluble fraction prepared from SDS-undoluble pellet by using the hot formamide exhibited a clear action of reducing sugars against the Anthronesulfate, while it exhibited no clear action against the ninhydrin. However, contrastly, the residue remainnning after extraction with formamide exhibited a clear action against ninhydrin and glucosamine was detected form the hydrolysate of residue by paper chromatography. From these results it is considered that the Sonic-supernatant fraction is mainly consisted of plasmic component of the cells. Other fractions, SDS-soluble, Formamide-soluble and Residue, should be consisted of plasma membrane, lipoplysaccharide and peptidoglycan of the cell, respectively.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.260-260
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• 1994

Mammalian pyruvate dehydrogenase complex(PDC) enzyme consists of multiple oopies of three major oligomeric enzymes-El, E2 E3. And protein X is one of the enzymatic constituents which is tightly bound to E2 subunit This complex enzyme is responsible for the oxidative decarboxylation of pyruvate producing of acetyl CoA which is a key intermediate for the entry of carbohydrates into the TCA cycle for its complete metabolic conversion to CO$_2$. And the overall activity of the complex enzyme is regulated via covalent nodification of El subunit by a El specific phosphatase ad kinase. Protein X has lipoyl moiety that undergoes reduction and acetylation during ezymatic reaction and has been known h be involved in the binding of E3 subunit to E2 core and in the regulatory activity of kinase. The purification of protein X has not been achieved majorly because of its tight binding to E2 subunit The E2-protein X subcomplex was obtained by the established methods and the detachment of protein X from E2 was accomplished in the 0.1M borate buffer containing 150mM NaCl. During the storage of the subcomplex in frozen state at -70$^{\circ}C$, the E2 subunit was precipitated and the dissociated protein X was obtained by cntrifegation into the supernatant The verification of protein X was accomplished by (1)the migration on SDS-PAGE, (2)acetylation by 〔2$\^$-l4/C〕 pyruvate, and (3)internal amino acid sequence analysis of tryptic digested enzyme.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.246-255
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• 1998

Ganoderan (GAN), an immunomodulating ${\beta}-glucan$ of G. lucidum, induces potent antitumor immunity in tumor-bearing mice. This study was set up to elucidate the ability of macrophage activation of GANs. GAN-treated Raw 264.7 macrophages showed enhanced production of nitric oxide (NO). The ability of GANs to produce NO was based on differences in chemical composition of GANs obtained from the mycelium on various carbon sources and mycelial fractionation. The highest NO production was observed in CW-AS-WS polysaccharide which was extracted from the mycelial wall. GAN-treated Raw 264.7 cells gave a 2-to 5-fold (24 hr) formation of NO levels compared with those treated with medium only. Partial removal of the protein in the extracellular GAN by TCA treatment did appreciably reduce its capacity to secrete NO. The mixture effect of GAN and LPS increased the nitric oxide secretion from RAW 264.7. The cell proliferation of GAN-treated Raw 264.7 cell tines inhibited as compared with its control. Of the culture supernatant of macrophage activated by GAN, the percentage of cytotoxicity against mouse leukemia L1210 cells was slightly dependent on the amount of NO in the culture supernatants of the activated-macrophages. These results indicate that the ${\beta}-glucan-related$ polysaccharides of the higher fungus activate macrophage and release nitric oxide. It also suggests that murine macrophages possess certain receptors for ${\beta}-anomeric$ glucans and play a critical role of ${\beta}-glucan-related$ tumor killing mechanism.