• Journal of Ginseng Research
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• v.10 no.2
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• pp.209-217
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• 1986

Yeast alcohol dehydrogenates and ginseng saponin interaction has been investigated to understand the non-specific enzyme stimulating effect of the saponin of Panax ginseng C.A. Meyer. It was confirmed that several amphiphiles such as sodium dodecyl sulfate(SDS), Triton X-100, sodium taurodeoxycholate (Na-TDC) as well as ginseng saponin mixture and purified ginseng glycosides lowered Km values of yeast alcohol dehydrogenase (ADH) for ethanol and NAD in the presence of the above amphiphiles suggesting that the surface activity of the amphiphiles might play a significant role in the ADH catalyzed reactions. Conformational change of yeast alcohol dehydrogenase in the presence of the above amphiphiles at their optimal concentration for the maximum activity was studied. Circular dichroism (C.D) spectrum of yeast ADH showed that the conformational change of the enzyme occurred in the presence of above amphiphiles. Fluorescence data also showed that the hydrophobic area increased in the presence of above amphiphiles. Examination of the interaction between ADH and ginseng saponin using radioactive saponin showed that there might be a very weak interaction between them. From the above results, it was concluded that the non-specific enzyme stimulating effect of the saponin might be due to the change of polarity of the enzyme solution in the presence of the saponin.

• Journal of Microbiology and Biotechnology
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• v.20 no.5
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• pp.881-888
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• 2010

solvent-tolerant bacterium strain, MH6, was isolated by hydrophilic organic solvent DMSO enrichment in the medium and identified as Serratia marcescens. The extracellular protease with novel organic-solvent-stable properties from strain MH6 was purified and characterized. The molecular mass of the purified protease was estimated to be 52 kDa on SDS-PAGE. The open reading frame (ORF) of the MH6 protease encoded 504 amino acids with 471 amino acid residues in the mature protease. Based on the inhibitory effects of EDTA and 1,10-phenathroline, the MH6 protease was characterized as a metalloproteinase. The enzyme activity was increased in the presence of $Ni^{2+}$, $Mg^{2+}$, and $Ca^{2+}$. The protease could also be activated by the nonionic surfactants Tween 80 (1.0%) and Triton X-100 (1.0%). The protease showed remarkable solvent stability in the presence of 50% (v/v) solutions of long-chain alkanes and long-chain alcohols. It was also fairly stable in the presence of 25% solutions of hydrophilic organic solvents. Owing to its high stability in solvents and surfactants, the MH6 protease is an ideal candidate for applications in organic catalysis and other related fields.

• Bulletin of the Korean Chemical Society
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• v.23 no.10
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• pp.1381-1391
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• 2002

The solvent sublation using ion pairs of metal-2-naphthoate(2-HNph) and tetrabutyl ammonium ($TBA^+$) ion has been studied for the concentration and determination of ultra trace Bi(III), In(III) and Tl(Ⅲ) ions in water samples. The partition coefficients ($K_p$) and the extraction percentages of 2-HNph and the ion pairs to methyl isobutyl ketone (MIBK) were obtained as basic data. After the ion pair $TBA^+$·M$(Nph)_4^-$ was formed in water samples, the analytes were concentrated by the solvent sublation and the elements were determined by GF-AAS. The pH of the sample solution, the amount of the ligand and counter ion added and stirring time were optimized for the efficient formation of the ion pair. The type and amount of optimum surfactant, bubbling time with nitrogen and the type of solvent were investigated for the solvent sublation as well. 10.0 mL of 0.1 M 2-HNph and 2.0 mL of 0.1 M $TBA^+$ were added to a 1.0 L sample solution at pH 5.0. After 2.0 mL of 0.2%(w/v) Triton X-100 was added, the ion pairs were extracted into 20.0 mL MIBK in a flotation cell by bubbling. The analytes were determined by a calibration curve method with measured absorbances in MIBK, and the recovery was 80-120%.

• BMB Reports
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• v.38 no.3
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• pp.343-349
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• 2005

Gamma-glutamyltransferase (GGT, EC 2.3.2.2) which hydrolyzes glutathione (GSH), is required for the maintenance of normal intracellular GSH concentration. GGT is a membrane enzyme present in leukocytes and platelets. Its activity has also been observed in human neutrophils. In this study, GGT was purified from Triton X-100 solubilized neutrophils and its kinetic parameters were determined. For kinetic analyses of transpeptidation reaction, $\gamma$-glutamyl p-nitroanilide was used as the substrate and glycylglycine as the acceptor. Apparent $K_m$ values were determined as 1.8 mM for $\gamma$-glutamyl p-nitroanilide and 16.9 mM for glycylglycine. The optimum pH of GGT activity was 8.2 and the optimum temperature was $37^{\circ}C$. It had thermal stability with 58% relative activity at $56^{\circ}C$ for 30 min incubation. L-serine, in the presence of borate, was detected as the competetive inhibitor. Bromcresol green inhibited neutrophil GGT activity as a noncompetetive inhibitor. The neutrophils seem to contain only the isoenzyme that is present in platelets. We characterized the kinetic properties and compared the type of the isoenzyme of neutrophil GGT with platelet GGT via polyacrylamide gel electrophoresis (PAGE) under a standart set of conditions.

• BMB Reports
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• v.37 no.4
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• pp.416-421
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• 2004

The antioxidant activities of NADH and of its analogue, 1,4-dihydro-2,6-dimethyl-3,5-dicarbethoxy-pyridine ($PyH_2$), were evaluated in vitro. NADH was found to be oxidized by the peroxyl radical derived from 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH) decomposition, in a pH-dependent manner. Both NADH and $PyH_2$ inhibited the peroxidation of egg yolk lecithin (EYL) liposomes, although $PyH_2$ was more effective than NADH when 2,2'-azobis-4-methoxy-2,4-dimethyl-valeronitrile (methoxy-AMVN) was employed to induce EYL liposome peroxidation. The antioxidant activities of NADH and $PyH_2$ were also evaluated by measuring their influences on 1,3-diphenylisobenzofuran (DPBF) fluorescence decay in the presence of peroxyl radicals. NADH and $PyH_2$ were much more effective at inhibiting DPBF quenching in Triton X-100 micelles than in liposomes. These results indicate that NADH can inhibit lipid peroxidation despite being hydrophilic. Nevertheless, membrane penetration is an important factor and limits its antioxidant activity.

• Archives of Pharmacal Research
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• v.19 no.6
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• pp.435-440
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• 1996

The effect of benzalkonium chloride on skin permeability of partially modified antisense phosphorothioate oligonucleotides (PS-ODN), which are designed as scar formation inhibitor, was investigated using Franz Diffusion Cell. When the concentration ratio of PS-ODN-quarternary ammonium salt complex is more than 1:100, the apparent partition coefficient (APC) of each complex was increased in the following order; tetraphenyl phosphonium chloride (TPP) < cetyltrimethyl ammonium bromide(CTAB) < benzalkonium chloride (BZ). The permeability of PS-ODN through the rat skin increased in the presence of BZ. The fluxs of PS-ODN with BZ were increased by addition of Pluronic F 68 or Triton X-100 to phosphate buffered saline (PBS), respectively. When the mole ratio of PS-ODN to BZ is 1:10, the fluxs penetrated of PS-ODN with BZ was greatest. The increase of the permeability in the presence of BZ might be due to the formation of lipophilic ion-pair complex between PS-ODN and BZ. By regulation of mole ratio of PS-ODN to BZ, the development of topical dosage forms using PS-ODN as scar formation inhibitor will be possible with minimal systemic exposure.

• Journal of Microbiology
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• v.40 no.2
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• pp.129-133
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• 2002

The fungus, Emericellopsis minima W2, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from a waste water sample. Production of the PHB depolymerase from E. minima W2 (PhaZ/ sub Emi/) was significantly repressed in the presence of glucose. PhaZ/ sub Emi/ was purified by column chromatography on Octyl-Sepharose CL-4B and Sephadex G-100. The molecular mass of the PhaZ/ sub Emi/), which consisted of a single polypeptide chain, was estimated to be 48.0 kDa by SDS-PAGE and its pI vague was 4.4. The maximum activity of the PhaZ/ sub Emi/ was observed at pH 9.0 and 55$\^{C}$. It was significantly inactivated by 1mM dithiothreitol, 2mM diisopropyl fluorphosphate, 0.1mM Tween 80, and 0.1 mM Triton X-l00, but insensitive to phenylmethylsulfonyl fluoride and N-ethylmaleimide. The PhaZ/ sub Emi/ efficiently hydrolyzed PHB and its copolyester with 30 mol% 3-hydroxyvalerate, but did not act on poly(3-hydroxyoctanoate). It also hydrolyzed p-nitrophenylacetate and p-nitrophenylbutyrate but hardly affected the longer-chain forms. The main hydrolysis product of PHB was identified as a dimer of 3-hydroxybutyrate.

• Journal of the Korean Society of Clothing and Textiles
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• v.37 no.8
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• pp.1107-1116
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• 2013

This study examined the use of cutinase and lipase to process cotton/polyester blend fabric. Optimum treatment conditions for cutinase and lipase were investigated for cotton/polyester blend fabric. The properties of enzyme-treated fabrics were evaluated and compared in optimal treatment conditions. In addition, the possibility to provide an enzymatic finishing on blend fabrics using mixed enzymes in a two-step process were studied. The weight loss of cotton/polyester blend fabrics with Triton X-100 was 0.8% and the dyeing property of blend fabrics with calcium chloride increased by a factor of 1.2. The use of two enzymes in combination with cutinase and lipase in the presence of auxiliaries resulted in a cotton/polyester blend fabric weight loss of 0.8%. In addition, the dyeing properties of cotton/polyester blend fabrics improved by a factor of 1.5 and the moisture regain of cotton/polyester blend fabrics improved by a factor of 1.16. However, no marked loss was observed in tensile strength. The surface morphology of cotton/polyester blend fabrics is modified through a two-enzyme treatment. The treatment of cotton/polyester blend fabrics with cutinase and lipase maintains cotton strength and improves the moisture regain of polyester fabrics.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.745-752
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• 2007

A $\beta$-glucosidase from the algal lytic bacterium Sinorhizobium kostiense AFK-13, grown in complex media containing cellobiose, was purified to homogeneity by successive ammonium sulfate precipitation, and anion-exchange and gel-filtration chromatographies. The enzyme was shown to be a monomeric protein with an apparent molecular mass of 52 kDa and isoelectric point of approximately 5.4. It was optimally active at pH 6.0 and $40^{\circ}C$ and possessed a specific activity of 260.4 U/mg of protein against $4-nitrophenyl-\beta-D-glucopyranoside$(pNPG). A temperature-stability analysis demonstrated that the enzyme was unstable at $50^{\circ}C$ and above. The enzyme did not require divalent cations for activity, and its activity was significantly suppressed by $Hg^{+2}\;and\;Ag^+$, whereas sodium dodecyl sulfate(SDS) and Triton X-100 moderately inhibited the enzyme to under 70% of its initial activity. In an algal lytic activity analysis, the growth of cyanobacteria, such as Anabaena flos-aquae, A. cylindrica, A. macrospora, Oscillatoria sancta, and Microcystis aeruginosa, was strongly inhibited by a treatment of 20 ppm/disc or 30 ppm/disc concentration of the enzyme.

• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.1087-1097
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• 2016

An intracellular lipase from Anoxybacillus flavithermus HBB 134 was purified to 7.4-fold. The molecular mass of the enzyme was found to be about 64 kDa. The maximum activity of the enzyme was at pH 9.0 and 50℃. The enzyme was stable between pH 6.0 and 11.0 at 25℃, 40℃, and 50℃ for 24 h. The K_m and V_max of the enzyme for pNPL substrate were determined as 0.084 mM and 500 U/mg, respectively. Glycerol, sorbitol, and mannitol enhanced the enzyme thermostability. The enzyme was found to be highly stable against acetone, ethyl acetate, and diethyl ether. The presence of PMSF, NBS, DTT and β-mercaptoethanol inhibited the enzyme activity. Hg²⁺, Fe³⁺, Pb²⁺, Al³⁺, and Zn²⁺ strongly inhibited the enzyme whereas Li⁺, Na⁺, K⁺, and NH₄⁺ slightly activated it. At least 60% of the enzyme activity and stability were retained against sodium deoxycholate, sodium taurocholate, n-octyl-β-D-glucopyranoside, and CHAPS. The presence of 1% Triton X-100 caused about 34% increase in the enzyme activity. The enzyme is thought to be a true lipase since it has preferred the long-chain triacylglycerols. The lipase of HBB 134 cleaved triolein at the 1- or 3-position.