• Journal of Microbiology and Biotechnology
• /
• v.26 no.6
• /
• pp.1115-1123
• /
• 2016

_L-Asparaginase (E.C. 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the _L-asparaginase gene (_L-ASPG86) in the genus Mesoflavibacter, which consists of a 1,035 bp open reading frame encoding 344 amino acids. Following phylogenetic analysis, the deduced amino acid sequence of _L-ASPG86 (_L-ASPG86) was grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The _L-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant _L-asparaginase (r-_L-ASPG86) showed optimum conditions at 37-40℃, pH 9. Moreover, r-_L-ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-_L-ASPG86 was 687.1 units/mg under optimum conditions (37℃, pH 9, and 5 mM MnSO₄).

• Microbiology and Biotechnology Letters
• /
• v.38 no.2
• /
• pp.183-189
• /
• 2010

In the course of screening for anti-acidosis and anti-diabetes agent from natural products, the inhibitory activities of Nepales plant extracts against microbial $\alpha$-amylase and $\alpha$-glucosidase were evaluated. Among the 100 different kinds of ethanol extracts, Cedrus deodara (Roxb.) G. Don and Myrica nagi Thunb showed strong inhibition activities against $\alpha$-amylase. The $IC_{50}$ values of C. deodara (Roxb.) G. Don, M. nagi Thunb and acarose, a commercial available anti-diabetes agent, were 44.5, 47.5 and $50.5\;{\mu}g/mL$, respectively. Considering the crude extract of C. deodara (Roxb.) G. Don, and M. nagi Thunb, these extracts have strong potentials as anti-acidosis or anti-diabates agent. In a while, Cleistocalyx operculatus (Roxb.) extract showed a good inhibition activity to $\alpha$-amylase and $\alpha$-glucosidase, even it was recently reported. The selected three extracts did not show any hemolysis activity against human red blood cell up to 1 mg/mL, and the inhibition activities were maintained by heat or acid treatment. Moreover, treatment of HCl (0.01N) for 1 h into C. operculatus (Roxb.) and M. nagi Thunb increased the inhibition activity from 50% to 70%. Our results suggest that C. deodara (Roxb.) G. Don, M. nagi Thunb, and C. operculatus (Roxb.) are potential as anti-acidosis and anti-diabetes agents.

• Journal of Dairy Science and Biotechnology
• /
• v.35 no.4
• /
• pp.221-231
• /
• 2017

This study aimed to investigate the physiological characteristics and anti-obesity effects of a newly isolated bacterium, Lactobacillus plantarum K6. L. plantarum K6 showed good ${\alpha}-amylase$ inhibitory activity ($96.78{\pm}3.29%$), ${\alpha}-glucosidase$ inhibitory activity ($92.55{\pm}9.62%$), and lipase inhibitory activity ($85.17{\pm}0.79%$), and the strain inhibited the adipocyte differentiation of 3T3-L1 cells ($27.4{\pm}1.4%$) when present at a concentration of $100{\mu}g/mL$. L. plantarum K6 was isolated from kimchi and its physiological characteristics were investigated. A comparison of the sensitivity of the isolate to 15 different antibiotics showed that L. plantarum K6 is highly sensitive to erythromycin and highly resistant to vancomycin, ampicillin, and polymyxin B. This strain also showed high arylamidase and ${\beta}-galactosidase$ activities. Moreover, it was relatively tolerant to bile acid and low pH, and displayed resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus, with rates of 51.8%, 42.4%, 61.6%, and 54.9%, respectively. No bio genic amines were produced. L. plantarum K6 also showed high adhesion activity to HT-29 cells compared to L. rhamnosus GG. These results demonstrate that Lactobacillus plantarum K6 has potential as a probiotic with anti-obesity effects.

• Korean Journal of Environmental Agriculture
• /
• v.34 no.3
• /
• pp.186-191
• /
• 2015

BACKGROUND: Carotenoids which are a major source of vitamin A are contributed to have great potential role in anti-carcinogenic effects and eyesight. Carotenoids which can not synthesize in human body are required for food supply. The objectives of this study are to investigate compositions and contents of pumpkin (Cucurbita spp.) germplasms based on their pulp color. METHODS AND RESULTS: Carotenoids were extracted with 0.2% ascorbic acid in ethanol and saponified with 80% potassium hydroxide. Insoluble compounds were extracted into hexane. A total of nine carotenoids (three xanthophylls and six carotenes) were identified from pumpkin germplasms using HPLC equipped with photodiode array detector (450 nm). Especially, lutein and ${\beta}$-carotenes were major compound in germplasms. Among isomers of ${\beta}$-carotene, all-trans-${\beta}$-carotene (16-27% of total carotenoids) was predominant compositions. The mean of total carotenoid contents was showed as brown (286.1 mg/100 g dw) > dark green (217.0) > orange (153.4) > primrose (85.8) > dark yellow (80.3). On the basis of carotenoid information, PLS-DA score plots showed different patterns by cluster in pumpkin germplasms. It was considered that these differences of phenotype were relative closely to genotype. CONCLUSION: This study indicated that dark color of pumpkin pulp was presented in high-level of biological pigments. It may contribute to develop potentially beneficial functional food ingredients.

• Korean Journal of Environmental Agriculture
• /
• v.27 no.1
• /
• pp.86-91
• /
• 2008

Dicamba(3,6-dichloro-2-methoxybenzoic acid) is used to control for pre and post-emergence of annual and perennial broad-leaf weeds. It is very soluble in water and highly mobile, acidic herbicide. So it is easily moved and detected in groundwater. Zerovalent iron(ZVI) has been used for the reductive degradation of certain compounds through amination of nitro-substituted compounds and dechlorination of chloro-substituted compounds. In this study, we investigated the potential of ZVI for the oxidative degradation of dicamba in water. The degradation rate of dicamba by ZVI was more rapidly increased in pH 3.0 than pH 5.0 solution. The degradation percentage of dicamba was increased with increasing amount of ZVI from 0.05% to 1.0%(w/v) and reached above 90% within 3 hours of reaction. As a result of identification by GC-MS after derivatization with diazomethane, we obtained three degradation products of dicamba by ZVI. They were identified 4-hydroxy dicamba or 5-hydroxy dicamba, 4,5-dihydroxy dicamba and 3,6-dichloro-2-methoxyphenol. 4-Hydroxy dicamba or 5-hydroxy dicamba and 4,5-dihydroxy dicamba are hydroxylation products of dicamba. 3,6-dichloro-2-methoxyphenol is hydroxyl group substituted compound instead of carboxyl group in dicamba. We also confirmed the same degradation products of dicamba in the Fenton reaction which is one of oxidation processes using ferric sulfate and hydrogen peroxide. But we could not find out the dechlorinated degradation products of dicamba by ZVI.

• Journal of Animal Science and Technology
• /
• v.60 no.11
• /
• pp.26.1-26.9
• /
• 2018

Background: This experiment aimed at assessing polyphenol-rich plant biomass to use in complete feed making for the feeding of ruminants. Methods: An in vitro ruminal evaluation of complete blocks (CFB) with (Acacia nilotica, Ziziphus nummularia leaves) and without (Vigna sinensis hay) polyphenol rich plant leaves was conducted by applying Menke's in vitro gas production (IVGP) technique. A total of six substrates, viz. three forages and three CFBs were subjected to in vitro ruminal fermentation in glass syringes to assess gas and methane production, substrate degradability, and rumen fermentation metabolites. Results: Total polyphenol content (g/Kg) was 163 in A. nilotica compared to 52.5 in Z. nummularia with a contrasting difference in tannin fractions, higher hydrolysable tannins (HT) in the former (140.1 vs 2.8) and higher condensed (CT) tannins in the later (28.3 vs 7.9). The potential gas production was lower with a higher lag phase (L) in CT containing Z. nummularia and the component feed block. A. nilotica alone and as a constituent of CFB produced higher total gas but with lower methane while the partitioning factor (PF) was higher in Z. nummularia and its CFB. Substrate digestibility (both DM and OM) was lower (P < 0.001) in Z. nummularia compared to other forages and CFBs. The fermentation metabolites showed a different pattern for forages and their CFBs. The forages showed higher TCA precipitable N and lower acetate: propionate ratio in Z. nummularia while the related trend was found in CFB with V. sinensis. Total volatile fatty acid concentration was higher (P < 0.001) in A. nilotica leaves than V. sinensis hay and Z. nummularia leaves. It has implication on widening the forage resources and providing opportunity to use forage biomass rich in polyphenolic constituents in judicious proportion for reducing methane and enhancing green livestock production. Conclusion: Above all, higher substrate degradability, propionate production, lower methanogenesis in CFB with A. nilotica leaves may be considered useful. Nevertheless, CFB with Z. nummularia also proved its usefulness with higher TCA precipitable N and PF. It has implication on widening the forage resources and providing opportunity to use polyphenol-rich forage biomass for reducing methane and enhancing green livestock production.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.37 no.4
• /
• pp.405-409
• /
• 2008

This study was performed to investigate the physiological activities of Viola mandshurica. Antioxidant activity was evaluated by measuring total phenolic contents, reducing power, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azino-di-2-ethyl-benzthiazoline sulphonate (ABTS) radical scavenging activity while anti-diabetic activity was measured by inhibition activities on ${\alpha}$-amylase and ${\alpha}$-glucosidase. V. mandshurica extracts were prepared by extracting with four different solvents (methanol, ethanol, acetone, and water). The methanol extract showed the highest total phenol content (34.49 mg/g gallic acid equivalents) among the extracts. The water extract showed the highest reducing power (0.454) at the concentration of $1,000{\mu}g$/mL. The acetone extract showed the most potent radical scavenging activity. DPPH and ABTS radical scavenging activity of the acetone extract at the concentration of $1,000{\mu}g$/mL were 21.13% and 43.53%, respectively. The inhibitory activity of acetone extracts against ${\alpha}$-amylase and ${\alpha}$-glucosidase showed more than 100% at the concentration of $1,000{\mu}g$/mL. The results indicate that V. mandshurica might have potential antioxidant and anti-diabetic activities.

• Polymer(Korea)
• /
• v.29 no.5
• /
• pp.501-507
• /
• 2005

Small intestinal submucosa (SIS) had been widely used as a biomaterial without immune rejection responses. SIS sponges prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). SIS powders dissolved in $3\%(v/v)$ acetic acid aqueous solution for 48hrs and freeze-dried. EDC solution ($H_2O$ : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100mM. In vitro, rat-BMSCs seeded in SIS sponges and induced the osteogenesis for 28 days. We have characterized the osteogenic potential of rat-BMSCs in SIS sponges by alkaline phosphatase activity(ALP), n assay, SEM and RT-PCR for osteogenic phenotype. In SEM, all morphology of SIS sponges was regular and showed interconnected pore structure. By RT-PCR analysis, we observed type I collagen expression. These results demonstrate osteogenic differentiation of rat-BMSCs. In conclusion, we confirmed that the morphology of surface, cross-section, and side of SIS sponges were highly porous with good interconnections between each pores, which can support the surface of cell growth, proliferation, and differentiation. This result indicates that SIS sponge is useful for osteogenesis of BMSCs.

• Korean Journal of Soil Science and Fertilizer
• /
• v.37 no.6
• /
• pp.371-377
• /
• 2004

Sulfate adsorption in forest soils is a process of sulfur dynamics playing an important role in plant uptake, cation movement, acid neutralization capacity and so on. The relationship between sulfate adsorption and some physicochemical properties of four forest soils was investigated. Extractable sulfate contents and sulfate adsorption capacity (SAC) in the forest soils varied much among study sites. Extractable sulfate contents were more in sub-surface soils with lower organic matter and greater Al and Fe oxides than in surface soils. The average contents of $Al_d$ and $Fe_d$ in the sub-surface soils were 8.49 and $12.45g\;kg^{-1}$, respectively. Soil pH, cation exchange capacity and clay content were positively correlated with the extractable sulfate contents and SAC. Organic carbon content, however, was negatively correlated with the extractable sulfate contents, implying the competitive adsorption of sulfate with soil organic matter. Considerably significant correlation was found between inorganic + amorphous Al and Fe oxides and the sulfate adsorption, but crystalline Al and other fractions of Fe oxide showed no correlation. Relatively close relationship between the adsorbed sulfates and soil pH, cation exchange capacity, or amorphous Al oxides indicates that the accelerated soil acidification may substantially reduce the potential for sulfate adsorption contributing to sulfur flux in forest ecosystems.

• Parasites, Hosts and Diseases
• /
• v.57 no.6
• /
• pp.671-680
• /
• 2019

Cathepsin D (CatD, EC 3.4.23.5) is a member belonging to the subfamily of aspartic endopeptidases, which are classified into the MEROPS clan AA, family A1. Helminth parasites express a large set of different peptidases that play pivotal roles in parasite biology and pathophysiology. However, CatD is less well known than the other classes of peptidases in terms of biochemical properties and biological functions. In this study, we identified 2 novel CatDs (CsCatD1 and CsCatD2) of Clonorchis sinensis and partially characterized their properties. Both CsCatDs represent typical enzymes sharing amino acid residues and motifs that are tightly conserved in the CatD superfamily of proteins. Both CsCatDs showed similar patterns of expression in different developmental stages of C. sinensis, but CsCatD2 was also expressed in metacercariae. CsCatD2 was mainly expressed in the intestines and eggs of C. sinensis. Sera obtained from rats experimentally infected with C. sinensis reacted with recombinant CsCatD2 beginning 2 weeks after infection and the antibody titers were gradually increased by maturation of the parasite. Structural analysis of CsCatD2 revealed a bilobed enzyme structure consisting of 2 antiparallel β-sheet domains packed against each other forming a homodimeric structure. These results suggested a plausible biological role of CsCatD2 in the nutrition and reproduction of parasite and its potential utility as a serodiagnostic antigen in clonorchiasis.