In our previous study, we isolated and characterized a 1-deoxynojirimycin (DNJ)-producing bacterium, Bacillus subtilis MORI, from chungkookjang, a Korean traditional food. B. subtilis MORI was subjected to ${\gamma}$-irradiation and the resulting bacteria were screened for increased DNJ production. A mutant was identified that produced 7.6 times more DNJ and named B. subtilis MORI 3K-85. In this study, the protein profiles of both strains were compared by one-dimensional and two-dimensional gel electrophoresis (1-DE and 2-DE, respectively) under both native and denaturing conditions. The 1-DE native-PAGE and 1-DE SDS-PAGE analyses identified 5 and 7 bands, respectively, that were found at higher concentrations in B. subtilis MORI 3K-85 than in B. subtilis MORI. Similarly, 2-DE analyses identified 20 protein spots which were found at higher concentrations in B. subtilis MORI 3K-85. The peptide mass profiles of these 20 proteins were analyzed by MALDI-TOF and compared with peptide sequences of B. subtilis and B. amyloliquefaciens in the MASCOT database. This screening suggested that three dehydrogenases, an aldolase, a synthetase, an isomerase, a reductase, and a peroxidase are elevated in B. subtilis MORI 3K-85. Based on this data, one or more of the elevated 8 enzymes might be related to the DNJ biosynthetic pathway.
A metabolic study has been conducted to investigate the conversion of pentachlorophenol(PCP) to water soluble metabolites in soybean and rice cell suspension cultures as well as in intact rice plants. PCP in plant cells was found to be exclusively transformed into water soluble metabolites. The relative rate of the metabolic conversion of PCP in decreasing order was soybean cultures > rice cultures > rice plants. Also observed was that, the older the cultures grown, the lower the conversion rate was. Primary water soluble metabolites isolated from both the 5 day old soybean and 8 day old rice cells were specifically hydrolyzed only by ${\beta}$-glucosidic linkage specific glucosidase, suggesting that the metabolites are ${\beta}$-glucose conjugates. The amount of glucose conjugates was increased with increasing time of incubation of PCP up to 24 hr in both soybean and rice cultures; Thereafter, it was decreased progressively. Most of the glucose conjugates were further metabolized to more polar conjugates in cells, but a portion of them was excreted into the culture medium.
International Journal of Industrial Entomology and Biomaterials
/
v.4
no.2
/
pp.93-100
/
2002
Cocoon production, which is a representative of traditional sericulture shifted into silkworm powder production in the spring of 1995. This, infect, signifies the change from the dress-centered textile business to the bio-industry and the functional resource industry. One of the most outstanding shifting is utilization of silkworm larvae for anti-diabetic agent. In Asian countries including Korea, silkworm powder derived from the domestic silkworm (Bombyx mori L.) has long been favored for anti-diabetic agent, but its efficacy was not tested until last decade by modern scientific methods. In this article, we reviewed the major researches on the silkworm powder as a blood glucose-lowering substance. After the beginning test of the efficacy of silkworm powder by a cooperative research between Department of Sericulture and Entomlogy, NIAST, RDA and Kyung Hee University, substantial data have been accumulated so far, In a serial experiment to select best condition, the fifth instar larvae prepared by freeze dry method turned out to have the best blood glucose-lowering effect. In the pharmacological experiment to understand the mechanism of silkworm powder in small intestine, the silkworm powder turned out to inhibit the activity of ${\alpha}$-glucosidase, by competitively binding to $\alpha$-type disaccharides. The animal experiment showed that the extract of silkworm powder prevents a rapid increase of blood glucose level after meal and prevents hunger and law blood glucose level during empty stomach. In the experiment to isolate the major component of silkworm powder, which exerts blood glucose-lowering effect, 1-deoxynojirimy-cin (DNJ) was eventually mass-purified, and it turned out that DNJ isolated from silkworm powder was excellent in its blood glucose-lowering effect. In the experiment to understand the personal difference of the efficacy of the silkworm powder, clinical candidates were divided on the basis of the criterion of traditional Chinese medicine: Tae-Yang, Tae-Um, So-yang, and So-Um. The result showed that silkworm powder has a tendency to reduce blood glucose level at fasting and at 2 hours after meal, and this trend was somewhat obvious in the Tae-Um body type. In summary, we reviewed scientific papers on the efficacy of silkworm powder and its purified DNJ as a blood glucose-lowering agent. These suggest that silkworm powder truly possesses blood glucose-lowering effect as documented in the traditional Chinese medicine, although further researches will be required to develop them as "medical" resource instead of functional food.
The edible barnyard millets(Echinochloa spp.) which have essentially vanished in the farmhouses environment and in agricultural germplasm were evaluated with the aim of restoration as a crop. The proximate components and mineral elements of milled millet were nutritionally similar or better than brown rice, and the vitamin contents of $B_1$ and $B_2$ exceeded those of rice by 1.3 times and 2.3 times, respectively. ${\beta}$-Carotene which is absent from brown rice was detected at levels ranging from 15~31 ${\mu}g$ in millet samples. Nine essential amino acids, including histidine and arginine and eight non-essential amino acids, such as aspartic acid were detected. The sum of all amino acids was determined to be IEC518>525>510 in the range of 69~106 mg/g. Analysis of physiological active substances via their electron donating ability(EDA) revealed values ranging from 3.4~8.2%, with the total polyphenol component being 51.1~69.4 mg/g and ${\alpha}$-glucosidase inhibition ability determined as 8.3~10.9%. In terms of agronomical characteristics and yields of barnyard millet, three millet varieties(IEC510, 518, and 525) were suitable as edible crops. IEC525 was selected as optimum variety for cultivation on the basis of nutritional ingredients, physiological active substances, and yield.
Kim Kyoung-Cheol;Kim Si-Wouk;Kim Myong-Jun;Kim Seong-Jun
Biotechnology and Bioprocess Engineering:BBE
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v.10
no.1
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pp.52-59
/
2005
The study was targeted to saccharify foodwastes with the cellulolytic and amylolytic enzymes obtained from culture supernatant of Trichoderma harzianum FJ1 and analyze the kinetics of the saccharification in order to enlarge the utilization in industrial application. T. harzianum FJ1 highly produced various cellulolytic (filter paperase 0.9, carboxymethyl cellulase 22.0, ${\beta}$-glucosidase 1.2, Avicelase 0.4, xylanase 30.8, as U/mL-supernatant) and amylolytic (${alpha}$-amylase 5.6, ${\beta}$-amylase 3.1, glucoamylase 2.6, as U/mL-supernatant) enzymes. The $23{\sim}98\;g/L$ of reducing sugars were obtained under various experimental conditions by changing FPase to between $0.2{\sim}0.6\;U/mL$ and foodwastes between $5{\sim}20\%$ (w/v), with fixed conditions at $50^{\circ}C$, pH 5.0, and 100 rpm for 24 h. As the enzymatic hydrolysis of foodwastes were performed in a heterogeneous solid-liquid reaction system, it was significantly influenced by enzyme and substrate concentrations used, where the pH and temperature were fixed at their experimental optima of 5.0 and $50^{\circ}C$, respectively. An empirical model was employed to simplify the kinetics of the saccharification reaction. The reducing sugars concentration (X, g/L) in the saccharification reaction was expressed by a power curve ($X=K{\cdot}t^n$) for the reaction time (t), where the coefficient, K and n. were related to functions of the enzymes concentrations (E) and foodwastes concentrations (S), as follow: $K=10.894{\cdot}Ln(E{\cdot}S^2)-56.768,\;n=0.0608{\cdot}(E/S)^{-0.2130}$. The kinetic developed to analyze the effective saccharification of foodwastes composed of complex organic compounds could adequately explain the cases under various saccharification conditions. The kinetics results would be available for reducing sugars production processes, with the reducing sugars obtained at a lower cost can be used as carbon and energy sources in various fermentation industries.
The biological activities of ethanol, ethanol: water(1 : 1v/v) and water extracts from Glycyrrhiza uralensis Fisch, glycyrrhizin and enzymatically hydrolyzed glycyrrhizin were compared. About 50% of the growth of MCF7, A549, Hep3B and AGS cells were inhibited in adding 1.0 g/L of the crude extracts, glycyrrhizin and enzymatically hydrolyzed glycyrrhizin. For example, the ethanol extract inhibited 76%, 66% in MCF7 and Hep3B cells by adding 1.0 g/L. For cytotoxicity on human normal liver cell(WRL-68), the crude extracts were scored as above 26%. For the result of antimutagenecity using CHO V79 cell, the crude extracts proved more effective than other samples. The growth of human immune B and T cells were enhanced up to $1.2{\sim}1.3$ times by adding the crude extracts. In inhibitory effect of ${\alpha}-glucosidase$ activity was showed that the ethanol extract, water extract and ethanol: water (1 : 1v/v) extract were appeared 65%, 68%, 62% in adding 1.0 g/L. The higher enhancement of glutathione -S-transferase activity was observed in the ethanol extract as 257% compared to the control in adding 1.0 g/L. From the results, the biological activities of the crude extracts were equivalent or higher than glycyrrhizin and enzymatically hydrolyzed glycyrrhizin.
To increase the functional properties of black soybean, we prepared the Chungkukjang with black bean at $40^{\circ}C$ after its germination according to the time course. The black soybean was soaked for 3 hr and dried three times with 2 hr interval. The germination ratio reached 50% for 12 hr germination and 80% for 24 hr, respectively. The optimum temperature of the germination was $18^{\circ}C$. The general composition of the germinated black soybean were not different from the control black soybean. During germination and growth, protease activity decreased slightly, whereas glucoamylase activity increased. On the other hand, content of isoflavone was increased to two times when the germinated leaf was 20 mm length. Absolute amount of the isoflavone was about 260 ${\mu}g$/g. When we estimated the sensory test of the Dua-chungkukjang, the color and mucoid formation was not different from traditional chungkukjang but bad flavor was decreased compared to traditional chungkukjang. And the same result was detected in the Dua-chungkukjang soup.
Journal of The Korean Society of Inherited Metabolic disease
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v.6
no.1
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pp.15-23
/
2006
Purpose: Glycogen storage disease type III (GSD-III), is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme is amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), which is responsible for the debranching of the glycogen molecule during catabolism. The disease has been demonstrated to show clinical and biochemical heterogeneity, reflecting the genotype-phenotype heterogeneity among different patients. In this study, we analyzed mutations of the AGL gene in three unrelated Korean GSD-III patients and discussed their clinical and laboratory implications. Methods: We studied three GSD-III patients and the clinical features were characterized. Sequence analysis of 35exons and part exon-intron boundaries of the AGLgene in patients were carried out by direct DNA sequencing method using genomic DNA isolated from patients' peripheral leukocytes. Results: The clinical features included hepatomegaly (in all patients), seizures (in patient 2), growth failure (in patients 1), hyperlipidemia (in patients 1 and 3), raised transaminases and creatinine kinase concentrations (in all patients) and mild EKG abnormalities (in patients 2). Liver transplantation was performed in patient 2due to progressive hepatic fibrosis. Administration of raw-corn-starch could maintain normoglycemia and improve the condition. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 with c.1510_1511insT (p.Y504LfsX10), and patient 3 with c.3416 T>C (p.L1139P) and c.l735+1 G>T (Y538_R578delfsX4) mutations. Except R428K mutation, 4 other mutations identified in3 patients were novel. Conclusion: GSD-III patients have variable phenotypic characteristics resembling GSD-Ia. The molecular defects in the AGL gene of Korean GSD-III patients were genetically heterogeneous.
The characteristics of crude enzyme produced from Trichoderma viride($O_2-1$) which isolated from half spoiled Locust acacia wood (Robinia Pseudacacia Linne) were examined in this paper. The results obtained were summarized as follows: 1) As a results of enzymatic action on several cellulose substrate it was found that there were some sorts of cellulase in crude enzyme. 2) The activity of C.M.C. ase and ${\beta}-glucosidase$ were decreased in accordance with increasing concentration of substrate, and filter paper saccharifying enzyme relatively increased in accordance with increasing concentration of substrate and enzyme in couse of time. 3) The optimal pH of each enzyme was 5.0 and the range of stability on pH generally from 3 to 6. 4) In disintegrating activity on filter paper, in decomposing activity on C.M.C. and $p-nitrophenyl-{\beta}-D-glucoside$, and in saccharifying activity on filter paper, the optimal temperatures were $50^{\circ}C.,\;60^{\circ}C,\;and\;65^{\circ}C$. 5) The order of stability on temperature was as follow; saccharifying activity on filter paper decomposing activity on C.M.C. disintegrating activity on filter paper>decomposing activity on $p-nitrophenyl-{\beta}-D-glucoside$. 6) The activity of the enzymes was inhibited with mercuric and silver ion, and activated with potassium.
Aronia (Black chokeberry, Aronia melanocarpa) belonging to the Rosaceae family, is native to eastern North America. Aronia contain high levels of flavonoids, mostly anthocyanins and proanthocyanidins, which are known as condensed tannins. The dominant proanthocyanidins in aronia are (-)-epicatechin and (+)-catechin. The concentration of proanthocyanidins in aronia is higher than in other berries, however due to the astringent taste it is not desirable for consumption. Therefore, the purpose of this study is to evaluate the effect of aronia on the reduction in tannins by yeast isolated from regional Jeotgal. We isolated strains of yeast with high ${\beta}$-glucosidase activity from Jeotgal, with the MTY2 strains exhibiting a reduction in final tannin concentration according to thin layer chromatography (TLC) analysis. MTY2 was confirmed as Kazachstania servazzii using an 18S rDNA sequence and named as K. servazzii MTY2. K. servazzii MTY2 showed most significant growth when K. servazzii MTY2 was cultured in a solution of 10% (w/v) glucose, 3% (w/v) tryptone and 0.1% (w/v) sodium chloride. According to the high performance liquid chromatography (HPLC) analysis, the (+) - catechin peak is present, but (-) - epicatechin peak was reduced at culture condition added with 10% glucose in medium.
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