• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.360-367
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• 1990

The several glycoside hydrolysing enzymes related to rutin degradation are found to be rhamnosidase, glucosidase and rutinosidase. Rutinosidase was purified to electrophoretic homogeneity from cell extracts of rutin-degrading strain, MT-57, which was identified as a Arthrobacter sp. Its molecular weight was estimated to be 42, 000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 40, 000 by gel filtration. The optimum pH for enzyme was found to be 7.5, and relatively stable in alkaline solution. The optimum temperature for enzyme was $45^{\circ}C$, being stable up to $50^{\circ}C$ for 20 min. The Bm value of enzyme for rutin was 0.5 $\mu \textrm m$. The enzyme activity was increased by the chelating agent such as EDTA, $NaN_3$, and 8-hydroxyquinoline, was strongly inhibited by $CO_{2+}, Ni^{2+}$, and $Cu^{2+}$. The enzyme had high substrate specificity in the rutinoside.

• Journal of Korea Soil Environment Society
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• v.1 no.2
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• pp.61-72
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• 1996

A series of batch and lab-scale continuous tests were conducted to optimize the design parameters for the full-scale in-situ soil flushing experiments. The cleaning abilities of the surfactant solutions of Tween 80, Triton X-100 and SDS were compared for the soil artificially contaminated by hydrophobic organic contaminants: n-dodecane, naphthalene and anthracene. Tween 80 and Triton X-100 were shown to be efficient for n-dodecane. SDS and Tween 80 were shown to be efficient for naphthalene and anthracene. At the end of each column test, the sorbed amount of surfactant to soil was also measured. Tween 80 was found to be the least adsorbed surfactant to soil. The flushing ability at flowrate of 7 ml/min, was hampered comparing to flowrate of 3 and 5 ml/min. Initial pH of the soil did not significantly affect the flushing efficiencies. Tween 80 was determined as the most harmless surfactant for the Gram(+) and Gram(-) bacteria.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.277-288
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• 2017

Rhizomucor miehei NRRL 5282 and Rhizopus oryzae NRRL 1526 can produce lipases with high synthetic activities in wheat bran-based solid-state culture. In this study, the purification and biochemical characterization of the lipolytic activities of these lipases are presented. SDS-PAGE indicated a molecular mass of about 55 and 35 kDa for the purified R. miehei and Rh. oryzae enzymes, respectively. p-Nitrophenyl palmitate (pNPP) hydrolysis was maximal at $40^{\circ}C$ and pH 7.0 for the R. miehei lipase, and at $30^{\circ}C$ and pH 5.2 for the Rh. oryzae enzyme. The enzymes showed almost equal affinity to pNPP, but the $V_{max}$ of the Rh. oryzae lipase was about 1.13 times higher than that determined for R. miehei using the same substrate. For both enzymes, a dramatic loss of activity was observed in the presence of 5 mM $Hg^{2+}$, $Zn^{2+}$, or $Mn^{2+}$, 10 mM N-bromosuccinimide or sodium dodecyl sulfate, and 5-10% (v/v) of hexanol or butanol. At the same time, they proved to be extraordinarily stable in the presence of n-hexane, cyclohexane, n-heptane, and isooctane. Moreover, isopentanol up to 10% (v/v) and propionic acid in 1 mM concentrations increased the pNPP hydrolyzing activity of R. miehei lipase. Both enzymes had 1,3-regioselectivity, and efficiently hydrolyzed p-nitrophenyl (pNP) esters with C8-C16 acids, exhibiting maximum activity towards pNP-caprylate (R. miehei) and pNP-dodecanoate (Rh. oryzae). The purified lipases are promising candidates for various biotechnological applications.

• The Korean Journal of Food And Nutrition
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• v.15 no.2
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• pp.104-111
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• 2002

The peptidyl prolyl sis-trans isomerase (PPIase, EC 5.2.1.8) from bacillus stearothermophilus was extracted from the cells treated with by lysozyme. PPIase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPIase was estimated as 18kDa by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0∼8.0. The enzyme was considerably stable after heat treatment at 60$\^{C}$ for 30minutes, and the enzyme was quite stable up to 65$\^{C}$. The presence of the PPIase in the refolding solution accelerated the isomerization rate of the assay peptide. PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-l primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPI-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus.

• Journal of Chest Surgery
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• v.43 no.6
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• pp.576-587
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• 2010

Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.

• Journal of Life Science
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• v.18 no.1
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• pp.84-90
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• 2008

Bacterial colonies which were able to oxidize the manganese were isolated from six soil samples in Byungchon area. Among them, one bacterial strain was selected for this study based on its high manganese oxidation activity. This selected bacterial strain was identified as Pseudomonas sp. MN5 through physiological-biochemical test and analysis of its 16s rRNA sequence. This selected bacterial strain was able to utilize fructose and maltose, but they doesn't utilizing various carbohydrates as a sole carbon source. Pseudomonas sp. MN5 showed a very sensitive to antibiotics such as kanamycin, chloramphenicol, streptomycin and tetracycline, but a high resistance up to mg/ml unit to heavy metals such as lithium, manganese and barium. Optimal manganese oxidation condition of Pseudomonas sp. MN5 was pH 7.5 and manganese oxidation activity was inhibited by proteinase K and boiling treatment. The manganese oxidizing protein produced by Pseudomonas sp. MN5 was purified by ammonium sulfate precipitation, HiTrap Q FF anion exchange chromatography and G3000sw $_{XL}$ gel filtration chromatography. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, three manganese oxidizing protein with estimated molecular weights of 15 kDa, 46.7 kDa and 63.5 kDa were detected. Also, it was estimated that manganese oxidizing protein produced by Pseudomonas sp. MN5 were a kind of porin proteins through internal sequence and N-terminal sequence analysis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.24 no.3
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• pp.6-16
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• 1998

Ceramides are currently emerging as the major skin care ingredients due to !heir barrier properties in the stratum corneum of the human skin. Thus, major cosmetic companies have developed synthetic ceramide analogs for their own use. In this study, several ceramide mimic compounds , new skin barrier lipids, were designed and synthesized, and their physical and biological properties were investigated to evaluate their skin care capability. Several structures were designed from the variation of hydrophobic alkyl chain and hydrophilic moiety by the use of molecular modeling software. The selected targets were synthesized, and their properties and activities were studied as the pure form, in the emulsion, or in the lamellar mixture containing cholesterol and fatty acid. Some compounds, such as 1,3-bis(N-(2-hydroxyethyl)-palmitoylamino)-2-hydroxypropane, enhanced the restoration of skin barrier damaged by SDS(sodium dodecyl sulfate), and by acetone treatment. The rate of restoration was comparable to that of natural ceramides. The synthesized compounds alleviated SDS induced skin irritation and facilitated lamellar phase liquid crystal formation. The treatment of 1,3-Dis(N-(2-hydroxyethyl)-palmitoylam ino)-2-hyd roxypropane on the acetone damaged skin revealed that the compound promoted the recovery of intercellular lipid lamellar structure of stratum corneum layer. The replacement of palmitoyl groups of the compound with shorter alkyl chain gave lower emulsion viscosity and liquid crystal density, suggesting easier formulation and poorer barrier activity. Most of the synthesized compounds were non-irritable in various toxicological tests proving that they can be safely introduced to the skin care formulations.

• Journal of the Korean Applied Science and Technology
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• v.18 no.4
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• pp.306-315
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• 2001

Our study is aimed at proposal of systematic verification method of molecular structure using measuring method of selective ionic determination and spectrometry on 34 kinds of surfactants such as sodium dodecyl sulfate(SDS) which are most widely used today. In the IR spectrum, unsaturated fatty acids reveal themselves by HC= at $3000{\sim}3020cm^{-1}$, and intensity of $720cm^{-1}$ depends on carbon length of alkyl group. Also ethylene oxide(EO) adducts exhibit weak characteristic bands by $-CH_{2}-CH_{2}-O$ at 1350, 1100 and $950cm^{-1}$. Isethionate can be distinguished from diester succinate by intensity ratio of 1740 and $1200cm^{-1}$ spectrums, the ratio of latter is close to 1 due to 2 carboxylate radical in diester succinate. Quaternary ammonium salts exhibit characteristic band of $C_{4}N^{+}$ at $1000-900㎝^{-1}$. In the case of dialkyl dimethyl ammonium salts in quaternary ammonium surfactants, the spectrum of $3000cm^{-1}$ by $N-CH_{3}$ collapses to a very weak band at $3020cm^{-1}$. In ammonium heterocyclic derivatives, pyridinium salts show characteristic bands at 1640 and $1460cm^{-1}$, while imidazolinium salts exhibit characteristic band at $1620-1610cm^{-1}$. In the characteristic spectrum at $1080-1050cm^{-1}$ on OH radicals of the alkyl esters, primary alcohol appears as weak band and the 2 bands show in almost same intensity when primary and secondary alcohols exist together in one molecule. Also, alkyl ester of polyhydric alcohols appears as various broad band.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.276-282
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• 2014

An accurate and sensitive analytical method was established for the simultaneous determination of synephrine and n-methyltyramine contents by ultra-performance liquid chromatography (UPLC) with a fluorescence detector (FLD). A 70:30 (v/v) mixture of 10 mM sodium dodecyl sulfate (SDS) and acetonitrile was used as the mobile phase. The coefficient of correlation ($r^2$) was 0.9999 for both synephrine and n-methyltyramine, and their limits of detection (LOD) were 0.02 and 0.01 mg/kg, respectively. The percentage recoveries for synephrine and n-methyltyramine were 96.4% and 100.9%, respectively, from bitter orange (Citrus aurantium) samples. The synephrine and n-methyltyramine contents were 38.07-118.21 mg/kg and 0.27-0.56 mg/kg, respectively, in the orange fruit samples, while they were 14.61-120.39 mg/kg and up to 3.34 mg/kg, respectively, in the tested commercial orange juice samples. The differences in synephrine and n-methyltyramine content between orange fruit and commercial orange juice were not significant (p<0.05). These results suggest that UPLC-FLD can be applied to develop an analytical method of quality control for commercial orange juice.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.317-325
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• 2001

For heuristic purposes, the relative ratio of urease contents inside and outside cells was surveyed using nine ureB+ strains of Helicobacter pylori. the ratio of the enzyme specific activity appeared to vary greatly between the various H. pylori strains, ranging from 0.5 to 2.5. Besides the above compartment, urease was also richly found in the membrane fraction, especially in either peripheral or integral form. The urease distribution across the H. pylori membrane was significantly influenced by the ambient pH; the specific activity of external urease was highest at pH 5.5 with a narrow plateau, whereas the internal specific activity was highest within a pH range of 4.5 to 6.5 with a broad plateau. These finding strongly suggest that H. pylori urease is secretory and responded to the external pH. However, at pH 4.0 or below, no urease activity was detected in either the internal or external compartment, although an increase in the color development with 2,4,6-trinitrobenzene sulfonate (TNBS) was observed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that these phenomena may be related to a specific proteolysis in certain proteins, including urease or ${\gamma}$-glutamyl transpeptidase. Interestingly, the effect of ammonium ions n alleviating the enzyme inactivation inside the H. pylori cells was remarkably similar to that of D-glucose. In addition, it would appear that the cation acted as a surrogate of not only $Na^+$ but also $K^+$ thereby increasing the H. pylori P-type ATPase activity. This is of great interest, as it implies that the urease action in H. pylori is indispensible at any locus.