• International journal of advanced smart convergence
• /
• v.7 no.2
• /
• pp.101-111
• /
• 2018

Celiac disease (CD) is an immune-mediated enteropathy of small intestine diagnosed in both childhood and adulthood. CD is caused by gluten, which produces gliadorphin during its digestion. The enzyme dipeptidyl peptidase-4 (DPP4) breaks gliadorphin down nevertheless the last tripeptide remains and eventually inhibits DPP4, thus slowing down its process. Therefore, the idea is to produce an additional DPP4 enzyme which is crucial. Consequently, the functional DPP4 gene was cloned into pCDNA3 intermediate (FLAG+DPP4) vector and finally a recombinant plasmid pSB1C3 (Andersons promoters+FLAG+DPP4) was constructed using synthetic biology. Normally, a DPP4 inhibitor is used as a cure for diabetes. Another important concern was overexpression of DPP4, which might lead to diabetes, accordingly the work was also performed for the regulation of the DPP4 gene expression. In this regard, three types of Anderson promoters (strong, moderate and weak) were utilized to study the control overexpression. This is the first report of idealistic trial for control the exogenous DPP4 gene-expression by molecular biologic tools.

• Nutritional Sciences
• /
• v.9 no.3
• /
• pp.159-166
• /
• 2006

This study was performed to investigate antioxidative and anti-aging action of extracts from Sancho (Zanthoxylum schinifolium) leaves. Two extracts were obtained by 80% methanol extraction followed by subsequent fractionation with methylene chloride (MC) and n-butanol (B) and fed at one or three levels to rats on normal level (5%) of fat (control) and high fat(20%) in diets. Male Sprague-Dawley rats weighing about 100 g were divided into ten groups such as control diet group(C), control diet+0.50%B group (CB), control diet+0.50%MC group (CMC), high-fat diet group (HF), high-fat diet+0.25%B group (HBL), high-fat diet+0.50%B group (HBM), high-fat diet+0.75%B group (HBH), high-fat diet+0.25%MC group (HMCL), high-fat diet+0.50%MC group (HMCM) and high-fat diet+ 0.75%MC group (HMCH) and fed each diet for four weeks. The effects of the extracts on antioxidant enzyme activities and indices of lipid peroxidation and aging were seen only in high fat diet groups. Hepatic superoxide dismutase and aryleaterase activities were not changed by Sancho extracts. But glutathione peroxidase, catalase and paraxonase activities were significantly restored by both MC and B at the level of 0.75% lipid peroxide which was increased by high fat diet was significantly reduced by B and MC at the level of 0.25% and over. Lipofuscin fluorescence and cabonyl value were increased by high fat diet were reduced by B and MC at the level of 0.5% and over. It is concluded that the Sancho extracts can be utilized as functional ingredients of health foods for reducing oxidative stress.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.1
• /
• pp.18-25
• /
• 2015

To understand the metabolism of flavonoid rhamnoglycosides by human intestinal microbiota, we measured the metabolic activity of rutin and poncirin (distributed in many functional foods and herbal medicine) by 100 human stool specimens. The average α-L-rhamnosidase activities on the p-nitrophenyl-α-L-rhamnopyranoside, rutin, and poncirin subtrates were 0.10 ± 0.07, 0.25 ± 0.08, and 0.15 ± 0.09 pmol/min/mg, respectively. To investigate the enzymatic properties, α-L-rhamnosidase-producing bacteria were isolated from the specimens, and the α-L-rhamnosidase gene was cloned from a selected organism, Bifidobacterium dentium, and expressed in E. coli. The cloned α-L-rhamnosidase gene contained a 2,673 bp sequcence encoding 890 amino acid residues. The cloned gene was expressed using the pET 26b(+) vector in E. coli BL21, and the expressed enzyme was purified using Ni²⁺-NTA and Q-HP column chromatography. The specific activity of the purified α-L-rhamnosidase was 23.3 µmol/min/mg. Of the tested natural product constituents, the cloned α-L-rhamnosidase hydrolyzed rutin most potently, followed by poncirin, naringin, and ginsenoside Re. However, it was unable to hydrolyze quercitrin. This is the first report describing the cloning, expression, and characterization of α-L-rhamnosidase, a flavonoid rhamnoglycosidemetabolizing enzyme, from bifidobacteria. Based on these findings, the α-L-rhamnosidase of intestinal bacteria such as B. dentium seem to be more effective in hydrolyzing (1 →6) bonds than (1 →2) bonds of rhamnoglycosides, and may play an important role in the metabolism and pharmacological effect of rhamnoglycosides.

• Food Science of Animal Resources
• /
• v.32 no.6
• /
• pp.706-712
• /
• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Applied Chemistry for Engineering
• /
• v.23 no.1
• /
• pp.100-104
• /
• 2012

Oligosaccharides production showed various biological activities in vivo like functional foods and industrial materials utilized available within many practical applications which have obtained from the degradation of alginate. Alginate is rich in the main component of seaweeds especially the brown algae. We investigated what degrading alginate from seaweeds to make alginate oligosaccharides can utilize in various fields using enzyme secreting Erwinia tasmaniensis. In this study, we observed an optimal culture condition of E. tasmaniensis, and characteristics of alginate lyase secreting E. tasmaniensis. These bacteria, E. tasmaniensis, were isolated from Nuruk. In this case, a suitable growth factor for E. tasmaniensis was culture it for 36 h in broth media on concentration of 1.0% (w/v) alginate. The enzyme showed the highest level of alginate lyase activity when cultured on broth media containing 1.0% (w/v) sodium alginate for 72 h. Optimal condition of pH, temperature and duration time for alginate lyase activity were found to be pH 6.0, $20^{\circ}C$ and 60 min, respectively.

• Journal of Life Science
• /
• v.11 no.5
• /
• pp.393-399
• /
• 2001

In order to improve functionality of kochujang which is one of the traditional foods in Korea, sea tangle powder(2, 4, 6 and 8% sea tangle powder on the glutinous rice weight basis) was added to the raw material of kochujang and then investigated the bacterial counts and enzyme activities with control kochujang during the fermentation at 3$0^{\circ}C$ for 120 days. Bacterial count was about 10$^4$ cfu/g at initial stage of fermentation and then maintained 10$^{8}$ cfu/g after 60 days of fermentation. $\alpha$-amylase activity was gradually reduced during fermentation periods, so the activity was lost almost at late of fermentation $\beta$-amylase activity was rapid increased until 30 days of fermentation and the rapid decreased at 60 days of fermentation and after 90 days was slightly decreased. Activities of acidic protease and neutral protease were increased until 30 days of fermentation and then these were shown irregularities decreased.

• Fisheries and Aquatic Sciences
• /
• v.25 no.6
• /
• pp.327-334
• /
• 2022

Marine resources have various biological activities and their constituents are more novel than those of land organisms. Several biologically active constituents have been found in marine organisms. Recently, many studies have reported that marine animals (MAs) can be used as functional ingredients in functional foods or nutraceutical due to their health benefits. However, no studies have extensively investigated the cosmeceutical activities of MAs extracts. Here, 70% ethanol extracts of 38 MAs were investigated for their activities of whitening and anti-aging properties for use as materials in novel cosmeceuticals. Anti-aging activities were determined by skin aging-related enzyme activities (anti-collagenase, anti-elastase, anti-hyaluronidase) and whitening activities (anti-tyrosinase, anti-3,4-dihydroxyl-L-phenylalanine [DOPA] oxidation) evaluated by colorimetric method. Among the 38 MAs, we found that Urechis unicinctus and Petrosia corticata extracts showed the strongest inhibitory effects against tyrosinase and DOPA oxidation, respectively. Our results additionally showed that Protankyra bidentata extract might provide a major source of anti-hyaluronidase and anti-elastase; meanwhile, anti-collagenase effects were similar in most MAs. Overall, these results suggest that extracts of marine animals have potential as a tyrosinase, collagenase, elastase, and hyaluronidase inhibitors. Taken together, MA resources could be considered as a novel cosmeceutical agent to be applied in cosmetic industry.

• Journal of Nutrition and Health
• /
• v.38 no.8
• /
• pp.626-636
• /
• 2005

This study was undertaken to examine effects of dietary intake of garcinia cambogia extract, soy peptide and L-carnitine mixture on body weight gain and obesity-related bio-markers in rats fed high-fat diet for 9 weeks with or without regular treadmill exercise. Forty 5-week-old male Sprague-Dawley rats were randomly divided into four groups; sedentary control group (SC), exercised control group (EC), sedentary formula-fed group (SF), and exercised formula-fed group (EF). The SC and EC rats were fed high-fat control diet (fat comprises$40\%$ of total caloris), and SF and EF rats were fed high-fat formula (composed of garcinia cambogia, soy peptide and L-carnitine) supplemented diet. Statistical analyses by two-way ANOVA indicated that the regular treadmill exercise significantly lowered cumulative body weight gain, total visceral fat mass, and epididymal, perirenal and retroperitoneal fat pad weights, and serum concentrations of total cholesterol and LDL + VLDL cholesterol, insulin, c-peptide and leptin. Feeding the formula also resulted in significant reductions in cumulative body weight gain and visceral fat pad weights, along with other related parameters including serum total and LDL + VLDL cholesterol levels, and hepatic enzyme activities involved in fatty acid synthesis. Statistical analyses by one-way ANOVA revealed that the formula consumption significantly improved body weight gain ($18\%$ reduction), total visceral fat weight ($20\%$ reductions), and serum total ($43\%$ reduction) and LDL + VLDL cholesterol ($54\%$ reduction) levels, as well as serum levels of insulin ($49\%$ reduction), and c-peptide ($41\%$ reduction) in sedentary rats, but failed to exhibit significant reductions in these indices in animals under treadmill exercise program. Taken together, these results suggest that the treadmill exercise per n exhibited significant improvements in body fat reduction and other related bio-markers, and so the formula consumption did not achieve a further significant reductions in these bio-markers in exercised rats. Nevertheless, animals fed the formula with regular exercise showed the most efficient weight reduction compared to other groups either fed formula without exercise or received regular exercise without dietary supplementation.

• Journal of Nutrition and Health
• /
• v.45 no.5
• /
• pp.429-436
• /
• 2012

This study was performed to examine the characteristics of protein of red crab (Chionoecetes japonicus) shell powder hydrolyzed by commercial proteases. Red crab shell was digested by commercial proteases, such as Protamex (P), Neutrase (N), Flavourzyme (F), Alcalase (A), Protease M (PM) and Protease A (PA). Protein yield analyzed by Biuret assay, absorbance at 280 nm and brix revealed that PA was the enzyme having the highest proteolytic activity. SDS PAGE showed that molecular weight of proteins produced by protease treatments was various and below 150 kDa. Combinational treatment of proteases (PA + P, PA + PM, PA + F, PA + A) was tried whether these increase protein hydrolysis from red crab shell powder compared to a PA single treatment. Soluble protein content was similar, but amino acid concentration by combinational treatments was higher than PA single treatment [PA + P 247.4 mg/g > PA + F (206.4 mg/g) > PA + A (133.4 mg/g) > PA + PM (59.1 mg/g) > PA (54.9 mg/g)]. Amino acid composition by combinational treatments was slightly different. Most abundant essential amino acids were phenylalanine, glycine, alanine, and leucine, whereas tyrosine and cystine were not detected.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.33 no.2
• /
• pp.78-84
• /
• 2016

The antioxidant activity of silkworm powder treated by proteolytic enzyme was investigated. Total protein content of silkworm power was assayed using BCA, Bradford assays and SDS-polyacrylamide gel electrophoresis (PAGE) with alkaline protease treatment conditions including temperature and pH. The optimum condition of alkaline protease treatment for silkworm powder was found to be $60^{\circ}C$ and pH 7. The alkaline protease treatment resulted in increased contents of free amino acids, total polyphenol and total flavonoid compared to control group. The silkworm hydrolysates showed excellent antioxidant activities in various in vitro models such as 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2 - azino-bis(3-ethylbenzthiazoline-6)-sulfonic acid (ABTS) radical scavenging activity. These results provide useful information for using silkworm powder as an ingredient in functional foods and for exploiting alkaline protease treatment to improve the extractability and bioactivity of a raw material.