• Korean Journal of Agricultural Science
• /
• v.25 no.1
• /
• pp.124-130
• /
• 1998

An alkalophilic bacterium, the strain AC-711 as a potent producer of alkaline cellulase, was selected among many isolates from soil environments. Morphological, physiological and chemical characteristics of the strain AC-711 suggested that it belongs to the genus Pseudomonas according to the Bergey's Manual of Systematic Bacteriology, however the G+C mol% (54.43) of its chromosomal DNA is lower than the normal values of the genus. The major cell wall fatty acids were determined as 15:0 and 17:0 anteiso. The production of alkaline CMCase by the strain was maximal when grown on the mediun containing 1% carboxymethyl cellulose, 0.1% $KH_2PO_4$, 0.02% $CoCl_2$, 0.02% Tween 80, 0.5% $Na_2CO_3$, 0.8% yeast extract, pH 10.3 at $30^{\circ}C$ for 3 days, and the most of enzyme was excreted into culture mediun.

• Applied Biological Chemistry
• /
• v.49 no.3
• /
• pp.221-226
• /
• 2006

In order to search anti-obesity agents, the methanol extracts of 155 herbal medicines were screened using with pancreatic lipase, which is involved in conversion of triglycerol to fatty acid. Among the tested medicinal plants, methanol extracts of Amsonia elliptica, Arecae pericarpium, Biota orientalis, Cinnamomum cassia, Curcuma aromatia, Elsholtzia ciliate, Glycyrrhiza uralensis, Mucunae Caulis, Rhus javanica, and Rubus coreanus showed potent inhibition at final concentration of $200\;{\mu}g/ml$ on pancreatic lipase activity. All of them were extracted into chloroform fraction. The relative inhibitory activities against pancreatic lipase by orlistat, the chloroform fraction of Arecae pericarpium and Cinnamomum cassia were 89, 80 and 80%, respectively. These results demonstrated that the screened medicinal plants could be develop as the effective lipase inhibitors in preventing and ameliorating obesity of human beings.

• Microbiology and Biotechnology Letters
• /
• v.36 no.4
• /
• pp.299-306
• /
• 2008

Human pancreatic lipase is a digestive enzyme which is synthesized in pancreas, secreted into small intestine, and there hydrolyze the fat in food. Pancreatic lipase protein composes of catalytic domain and colipase-binding domain. In this research, the gene segments corresponding to total protein, catalytic domain, and co lipase-binding domain were cloned by PCR method, inserted into an expression vector, and then used to transform Escherichia coli BL21 (DE3). The recombinant proteins produced were purified and injected intramuscularly three times into laying hens. The egg yolk antibodies (IgY) were obtained from the egg yolks and tested for their antibody titer. Among three IgY, the IgY against colipase-binding domain showed the highest antibody titer. All three IgY had inhibitory effects on the porcine pancreatic lipase. Among them, the IgY against colipase-binding domain showed the highest inhibition effects. The fat diet with corn oil and IgY was administrated to the experimental rats and their blood compositions were examined with time course. The triglyceride concentration of treated rats was decrease meaningfully when compared with those of control rats. This suggested that the IgY against colipase-binding domain antigen inhibited pancreatic lipase in vivo.

• Korean Journal of Food Science and Technology
• /
• v.50 no.5
• /
• pp.528-534
• /
• 2018

The current study investigated the effects of enzyme treatment on black rice (Oryza sativa L.) aleurone layer extracts. Different enzymes (lipase, lecitase ultra and lipopan 50BG) were used to test anti-oxidant and anti-inflammatory activities in vitro. The antioxidant activities of enzyme treated or non-enzyme treated extracts of black rice bran were evaluated via 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity. Lipase treated extracts of black rice bran showed higher antioxidant activity compared to that of non-enzyme treated extracts. Anti-inflammatory activities of enzyme treated black rice bran extracts on nitrite production and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) secretion, were tested using a nitric oxide (NO) colorimetric assay kit and an enzyme-linked immunosorbent assay (ELISA) kit. The ethanolic extract of enzyme treated black rice bran decreased the levels of nitric oxide production and pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$, in a lipopolysaccharidestimulated RAW 264.7 cell culture. These findings indicate that enhanced anti-oxidant and anti-inflammation activities of the ethanolic extracts of enzyme treated black rice (Oryza sativa L.) aleurone layers, may be attributed to molecular conversion of ingredients in enzyme catalyzed reactions.

• Microbiology and Biotechnology Letters
• /
• v.46 no.3
• /
• pp.234-243
• /
• 2018

The Antarctic Ocean contains numerous microorganisms that produce novel biocatalysts that can have applications in various industries. We screened various psychrophilic bacterial strains isolated from the Ross Sea and found that a Croceibacter atlanticus strain (Stock No. 40-F12) showed high lipolytic activity on a tributyrin plate. We isolated the corresponding lipase gene (lipCA) by shotgun cloning and expressed the LipCA enzyme in Escherichia coli cells. Homology modeling of LipCA was carried out using the Spain Arreo lake metagenome alpha/beta hydrolase as a template. According to the model, LipCA has an ${\alpha}/{\beta}$ hydrolase fold, Gly-X-Ser-X-Glymotif, and lid sequence, indicating that LipCA is a typical lipase enzyme. Active LipCA enzyme was purified fromthe cell-free extract by ammonium sulfate precipitation and gel filtration chromatography. We determined its enzymatic properties including optimum temperature and pH, stability, substrate specificity, and organic solvent stability. LipCA was immobilized by the cross-linked enzyme aggregate (CLEA) method and its enzymatic properties were compared to those of free LipCA. After cross-linking, temperature, pH, and organic solvent stability increased considerably, whereas substrate specificities did not changed. The LipCA CLEA was recovered by centrifugation and showed approximately 40% activity after 4th recovery. This is the first report of the expression, characterization, and immobilization of a C. atlanticus lipase, and this lipase could have potential industrial application.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.41 no.12
• /
• pp.1677-1685
• /
• 2012

The autolysate and hydrolysate of a common squid liver, Todarodes pacificus, were prepared. Autolysis (liver ratio, pH, temperature) and Protamex-treated hydrolysis (pH, temperature, ratio of protease to liver) conditions were optimized by response surface methodology using central composite design for under 1 hr of hydrolysis time. The desirability profile indicated that maximum DH could be achieved at a squid liver of 93.5%, pH 6.4, and $47^{\circ}C$ in autolysis, while that of Protamex-treated hydrolysis did at a Protamex-to-squid liver level of 0.33%, pH 6.0, and $55^{\circ}C$. Three amino acids, proline, cysteine, and methionine, were not detected in the total amino acid composition of the Protamex-treated hydrolysate, while they were detected in the free amino acid composition. Cadmium was $8.32{\pm}0.03$ mg/100 g-powder for raw, $3.56{\pm}0.02$ mg/100 g-powder for the autolysate, and $13.26{\pm}0.04$ mg/100 g powder for the Protamex-treated hydrolysate. The major molecular weight ranged from 1.0 to 1.5 kDa for the autolysate and from 210 to 470 Da for the Protamex-treated hydrolysate. Food functionalities of the autolysate, such as surface hydrolphobicity, emulsion activity index, emulsion stability, water, and fat adsorption, were similar to the Protamex-treated hydrolysate. Both the autolysate and Protamex-hydrolysate showed high inhibitory activities on the angiotensin-I converting enzyme. Cell toxicity against the HepG2 cell line was not detected in the autolysate or the Protamex-treated hydrolysate by 200 ${\mu}g/mL$.

• Journal of Mushroom
• /
• v.13 no.1
• /
• pp.56-62
• /
• 2015

A lipase producing bacterium was isolated from button mushroom bed, which showing high clear zone on agar media containing Tributyrin as the substrate. The strain was identified as Burkholderia cepacia by analysis of 16S rDNA gene sequence. Crude lipase (CL) was partially purified from 70% ammonium sulfate precipitation using the culture filtrate of B. cepacia. Immobilized lipases were prepared by cross-linking method with CL from B. cepacia and Novozyme lipase (NL) onto silanized Silica-gel as support. Residual activitiy of the immobilized CL (ICL) and immobilized NL (INL) was maintained upto 61% and 72%, respectively. Biodiesel (Fatty acid methyl ester, FAME) was recovered by transesterification and methanolysis of Canola oil using NaOH, CL and ICL as the catalysts to compare the composition of fatty acids and the yield of FAME. Total FAME content was NaOH $781mg\;L^{-1}$, CL $681mg\;L^{-1}$ and ICL $596mg\;L^{-1}$, in which the highest levels of FAME was observed to 50% oleic acid (C18:1) and 22% stearic acid (C18:0). In addition, the unsaturated FAME (C18:1, C18:2) decreased, while saturated FAME (C16:0, C18:0) increased according to increasing the reaction times with both CL and ICL, supporting CL possess both transesterification and interesterification activity. When reusability of ICL and INL was estimated by using the continuous reaction of 4 cycles, the activity of ICL and INL was respectively maintained 66% and 79% until the fourth reaction.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.9
• /
• pp.1321-1327
• /
• 2011

The microorganism producing an invertase (E.C. 3.2.1.26) was isolated from wine and tentatively identified as Saccharomyces cerevisiae by cellular fatty acid analysis. The invertase was purified to homogeneity by ammonium sulfate precipitant, dialysis, ion-exchange chromatography on DEAE-Sephadex A-50, and gel chromatography on Sephadex G-200 from the culture supernatant of Saccharomyces cerevisiae JS59. The specific activity and the purification fold of the purified invertase were 7620.9 unit/mg protein and 13.9, respectively. The molecular weight of the purified invertase was estimated to be 38.5 kDa by SDS-PAGE. The optimum pH and temperature for the invertase activity were pH 5 and $55^{\circ}C$, respectively. The invertase activity was relatively stable at pH 4~6 and temperature $55^{\circ}C$. The activity of invertase was inhibited by $Ag^{2+}$ and $Hg^{2+}$, but on the contrary, activated by $Co^{2+}$ and $Mn^{2+}$. Michaelis constant ($K_m$) for invertase reaction in sucrose solution was 11.5 mM. TLC analysis of the products produced in sucrose solution during invertase reaction showed the progressive presence of glucose and fructose in accordance with sucrose hydrolysis.

• Journal of Life Science
• /
• v.24 no.7
• /
• pp.798-803
• /
• 2014

Cathepsin D (CtsD), an aspartyl peptidase, is involved in apoptosis, resulting in the release of cytochrome C from mitochondria in cells. Here, we investigated microRNA regulation of CtsD expression in 3T3-L1 cells First, we observed the expression of CtsD in cells in response to doxorubicin (Dox). As expected, the level of CtsD mRNA was increased in 3T3-L1 cells exposed to Dox in a dose-dependent manner. Cellular viability of ectopically expressed CtsD cells was also decreased. Next, we used the miRanda program to search for particular microRNA targeting CtsD. MiR-145 was selected as a putative controller for CtsD because miR-145 had a high mirSVR score. In a reporter assay, the luciferase activity of cells containing the CtsD 3'-UTR region was decreased in cells transfected with miR-145 mimic compared to that of a control. The level of CtsD expression was down-regulated in preadipocytes ectopically expressing miR-145 and up-regulated by an miR-145 inhibitor. Cells also suppressed miR-145 expression when exposed to Dox. The miR-145 inhibitor reduced the cellular viability of 3T3-L1 cells. Taken together, these data suggest that miR-145 regulates CtsD-mediated cell death in adipocytes. These findings may have valuable implications concerning the molecular mechanism of CtsD-mediated cell death in obesity, suggesting that CtaD could be a useful therapeutic tool for the prevention and treatment of obesity by regulating fat cell numbers.

• Applied Biological Chemistry
• /
• v.28 no.4
• /
• pp.226-232
• /
• 1985

Changes in lipid and fatty acid composition in Korean native Meju were investigated at one week interval over 6 weeks of fermentation. For the systematically, salicic acid column chromatography and gas liquid chromatography were used. Following results were obtained. 1. The lipid fraction obtained from the soared soybean and the cooked soybean were mainly composed of $72.47%{\sim}92.35%$ of neutral lipid, phospholipid and glycolipid were 4.64% and 4.88%, respectively. During fermentation period, lipid content decreased to 80.59%, but glycolipid and phospholipid contents increased. 2. The triglyceride contents of nonpolar lipids prepared from the cooked soybean and the soaked sobean was 89.66% and 87.83% respectively. Free fatty acid, diglyceride and sterol contents increased during fermentation, whereas triglyceride content decreased. 3. Lipids extracted from the soaked soybean and the cooked soybean were composed of 54.58% linoleic acid, $22{\sim}20%$ oleic acid, $10{\sim}12%$ palmitic acid and $3{\sim}5%$ stearic acid. 4. During the Korean native Meju fermentation in palmitic acid decreased from the second week and stearic acid through $3rd{\sim}4th$ week. Oleic acid and linoleic acid content decreased gradually, but linolenic acid content increased. 5. During the fermentation, myristic acid content of glycolipid fraction increased. Lipase activity reached to the maximum at the 3rd week.