• The Korean Journal of Food And Nutrition
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• v.7 no.2
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• pp.128-136
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• 1994

Mackerel muscle protein hydrolysates, which were prepared from defatted mackerel meal by proteases such as complex enzyme, alcalase, bromelain, pancrease, pepsin, w-chymotrypsin, trypsin and papain, were tested for the antioxidative action against linoleic acid. Among proteases tested, the hydrolysates obtained from the treatment of complex enzyme, bromelain and alcalase showed higher antioxidative effects. Also, the hydrolysates showed the synergistic effects with o-tocopherol and the inhibitory effects for peroxidation of metal ions(Fe3+, Cua+) From the profiles of fractionation of the hydrolysates with Bio-gel P-2 column, the most active fractions, part I(complex enzyme-derived) and part e(bromelain-derived), had below MW 1,400 and the antioxidative effects were closely related to the binding capacity with metal ion(Cua+). Amno acid composition of the part I was abundant in histidine, arginine, phenylalanine and lysine, and the part e was abundant in lysine, glutamic acid and leucine.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.1
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• pp.97-106
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• 1984

The Antarctic krill, Euphausia superba, is drawing attention over the world as the largest source of unutilized proteins in the ocean. For the use of krill as a human food, processing conditions of krill sauce by autolysis and/or commercial proteolytic enzyme digestion were examined. The krill was chopped and mixed with equal weight of water, and hydrolyzed by autolysis and/or commercial proteolytic enzyme digestion. The optimal conditions for hydrolysis of krill were $52.5^{\circ}C$, pH 7.0-7.5, 3 hours by autolysis, $52.5^{\circ}C$, pH 6.3, 3hours by bromelain (0.5 %) digestion, and $52.5^{\circ}C$, pH 7.0-7.5, 3 hours by commercial complex enzyme (5 %) digestion, respectively The maximum hydrolyzing rate of protein were 83.2 % by autolysis, 89.7 % by bromelain digestion, 92.7 % by commercial complex enzyme digestion. After krill meat hydrolyzed by autolysis at optimum condition, inactivated at $100^{\circ}C$ for 20 minutes and filtered with Buchner funnel. Two kinds of products were prepared with krill hydrolysate and preservatives: one contained 10 % of sodium chloride and 0.06 % of benzoic acid and the other 10 % of sodium chloride and 3 % of ethyl alcohol. These products were filled in the sterilized glass bottle and sealed. The pH, volatile basic nitrogen, amino nitrogen, color value (L, a and b values) and viable counts of bacteria were determined during storage at $37^{\circ}C$. The results showed that the products could be preserved in good condition during one month at $37^{\circ}C$. As a method to reduce the sodium level in krill sauce, it is convinced that sodium chloride could be replaced half in partially by potassium chloride. In the products prepared from krill by autolysis, bromelain or commercial complex enzyme digestion, hypoxanthine and 5'-IMP were abundant among the nucleotides and their related compounds as 15.3-20.4 ${\mu}mole/g$, dry solid, 2.2-2.5 ${\mu}mole/g$, dry solid, respectively. The abundant free amino acids were lysine, leucine, proline, alanine and valine. The contents of these amino acids were 67.4 %, 69.4 %, 69.8 % of the total free amino acids of each products. And TMAO, betaine and total creatinine were low in contents. The flavor of krill sauce prepared from krill by autolysis or enzyme digestion was not inferior to that of traditional Kerean soy sauce by sensory evaluation.

• BMB Reports
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• v.28 no.1
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• pp.12-16
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• 1995

The branched-chain ${\alpha}$-keto acid dehydrogenase (BCKAD) complex is a rate limiting enzyme which catalyzes the oxidative decarboxylation of branched-chain ${\alpha}$-keto acids. Numerous studies have suggested that BCKAD is subject to covalent modification in vitro via phosphorylation and dephosphorylation, which are catalyzed by a specific kinase and phosphatase, respectively. The biggest difficulty in the assay of BCKAD activity is to arrest the interconversion between the active and inactive forms. BCKAD activity was determined from fresh rat heart and liver tissues using homogenizing and assay buffers containing inhibitors of phosphatase and kinase. The results suggest that a radiochemical assay using ${\alpha}$-keto[1-$^{14}C$]-isovalerate as a substrate for the enzyme can be applied as a reliable method to determine in vitro enzyme activity with arrested interconversion between the active and inactive forms of the BCKAD complex.

• Journal of Animal Science and Technology
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• v.53 no.2
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• pp.113-118
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• 2011

This study was conducted to evaluate effects of enzyme complex on growth performance, nutrient digestibility, blood profiles and feed cost in growing pigs. Ninety-six pigs [(Landrace ${\times}$ Yorkshire) ${\times}$ Duroc, $22.96{\pm}0.79$ kg average initial body weight] were used in 42d growth assay. Dietary treatments included:1) HC (high energy and nutrient density diet), 2) CON (control, basal diet), 3) CE1 (CON + 0.05% enzyme complex) and 4) CE2 (CON + 0.1% enzyme complex). Four pigs were allotted per pen with six replicate pens per treatment by completely randomized design. The ADG was higher in CE1 and CE2 treatments than CON treatment (P<0.05). The ADFI was linearly increased by CE treatments compared to HC treatment. The CE1 treatment had highest DM, N and GE digestibility (P<0.05). Digestibility of DM, N and GE were quadratic enhanced by enzyme complex level. No differences were found among treatments for creatinine and BUN. The enzyme complex treatments (CE1 and CE2) showed lower feed cost/body weight gain than HC treatment. In conclusion, enzyme complex can improve ADG and reduce feed cost/body weight gain when low energy diet was used. Furthermore adding 0.05% enzyme complex had highest nutrient digestibility.

• The Korean Journal of Food And Nutrition
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• v.28 no.4
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• pp.628-634
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• 2015

The ground red pepper puree was treated with cellulase (C treatment), pectinase (P treatment), amylase (A treatment) and/or enzyme complex (CP, CA, PA and CPA treatment) for 2~8 hours to improve the yield and bioactivity of extracts. And physicochemical and sensory properties of red pepper extracts were evaluated. The extraction yield of the control was 38.84%, which was lower than those of the enzyme treatments. And extraction yields of enzyme complex treatments were higher than that of single enzyme treatments. Especially, extraction yield was increased to 74.37% by cellulase + pectinase + amylase complex treatment (CPA treatment). The soluble solid and reducing sugar contents were higher in the extracts treated with enzymes compared with the control. CA and CPA treatment showed the highest soluble solid and reducing sugar contents. No significant changes in lightness, redness and yellowness of the control and the samples by enzyme treatments were observed during 2~8 hours experiments. The sensory evaluation results revealed that panelists preferred the extracts with enzyme treatments to the control. Therefore, enzyme treatment for red pepper extracts is a good method to improve the yield and sensory properties.

• Applied Microscopy
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• v.17 no.2
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• pp.55-71
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• 1987

To investigate the effects of lead on the electron transport system and ultrastructure of mouse kidney mitochondria, various lead acetate concentrations were treated in vitro and respiration rate, enzyme activities were measured. Ultrastructural changes at state IV respiration were also observed. To compare with in vivo experiments, mouse were injected intraperitoneally of 100 mg lead acetate per kg body weight and state IV respiration rate and enzyme activities were measured. Ultrastructure of renal proximal tubular cells were also observed. In in vitro treatement, decreased state IV respiration, decreased enzyme activities, ruptured membranes and inhibition of condensed to orthodox transformation were observed. In in vivo treatment, decreased state IV respiration and decreased enzyme activities were observed after 24 hrs of i.p. injection. Cytochrome c oxidase activity showed twice the inhibition compared to NADH-CoQ reductase activity at 24 hrs. Continuous decreased state IV respiration was observed after 48 and 72 hrs of injection, however, the enzyme activities were increased to control level. Lead-protein complex which probably inhibits the toxic effects of lead appeared. To conclude, dominant effect of lead on the electron transport system appeared at cytochrome c oxidase activity, and the increased enzyme activities may be a result of appearance of lead-protein complex.

• Interdisciplinary Bio Central
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• v.4 no.4
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• pp.13.1-13.6
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• 2012

The advanced glycation endproducts receptor (AGER) is a multiligand signal transduction receptor. One of its ligands, S100b molecules activates vascular smooth muscle cells and endothelial cells via its receptor, thus triggering activation of signaling cascades and generation of cytokines and proinflammatory molecules. Ubiquitin-conjugating enzyme Ubc9 is an E2 conjugating enzyme that transfers the activated small ubiquitin-related modifier to protein substrates, and thus it plays a critical role in SUR-Mylation-mediated cellular pathways. Previous studies have shown that both AGE-R and Ubc9 play roles in diverse cellular signaling pathways. However, until recently, little attention has been paid to interactions between AGE-R and Ubc9. In this study, sequence database searches allowed us to identify a potential interaction motif between AGE-R and Ubc9. The subsequent biochemical and molecular biological analysis suggested that there may be specificity in AGE-R and Ubc9 complex signaling in atherosclerosis and cancer cells in a cell-type specific manner. Although the determinant for specificity in AGE-R and Ubc9 complex signaling in cancer cells and atherosclerosis is yet to be determined, this study provides the basis to develop a specific therapeutic application of AGE-R, SURM (small ubiquitin-related modifier)-1, and Ubc9 complex activation pathways in atherosclerosis, diabetes, cancer and inflammatory diseases.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1636-1643
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• 2014

3-Hydroxybutyryl-CoA dehydrogenase is an enzyme that catalyzes the second step in the biosynthesis of n-butanol from acetyl-CoA, in which acetoacetyl-CoA is reduced to 3-hydroxybutyryl-CoA. To understand the molecular mechanisms of n-butanol biosynthesis, we determined the crystal structure of 3-hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum (CbHBD). The monomer structure of CbHBD exhibits a two-domain topology, with N- and C-terminal domains, and the dimerization of the enzyme was mostly constituted at the C-terminal domain. The mode of cofactor binding to CbHBD was elucidated by determining the crystal structure of the enzyme in complex with $NAD^+$. We also determined the enzyme's structure in complex with its acetoacetyl-CoA substrate, revealing that the adenosine diphosphate moiety was not highly stabilized compared with the remainder of the acetoacetyl-CoA molecule. Using this structural information, we performed a series of site-directed mutagenesis experiments on the enzyme, such as changing residues located near the substrate-binding site, and finally developed a highly efficient CbHBD K50A/K54A/L232Y triple mutant enzyme that exhibited approximately 5-fold higher enzyme activity than did the wild type. The increased enzyme activity of the mutant was confirmed by enzyme kinetic measurements. The highly efficient mutant enzyme should be useful for increasing the production rate of n-butanol.

• Archives of Pharmacal Research
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• v.15 no.3
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• pp.229-233
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• 1992

The pKa value of histidine-51 residue was determined by the pH dependency of contents of NADH bound to the active site in the orse liver alcohol dehydrogenase and % inactivation with diethyl pyrocarbonate treatment of the enzyme. The pKa for His-51 was -7.15 in the ternary complex and -6.7 in the enzyme itself.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.524.3-524
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• 1986

Carboxymethyl cellulase (CMCase) produced by cloned B. megaterium was found to contain 5.2% carbohydrate but no metal ion. The enzyme was isoelectric at pH 7.23 and was high is basic amino acids. The N-terminal of the enzyme was glutamic acid. The cellulolytic activity of this enzyme was extended to the small molecular substrates such as from cellotriose to cellopentaose. In additon, the enzyme showed transglycoslation activity. The pK values of the enzyme we estimated to be 4.4 and 6.7, andthat of the enzyme-substrate complex were 4.2 and 7.2, respectively. The enzyme was not affected by the treatment with iodoacetic acid, but the modification of enzyme with carbodiimide and diethyl pyrocarbonate resulted in a marked loss of the enzyme activity. These results suggest that the active site of enzyme essentially contains carboxylic and imidazole group of amino acid residues.