• Journal of the Korean Society of Food Science and Nutrition
• /
• v.19 no.6
• /
• pp.625-635
• /
• 1990

The purpose of this study was focused on investigation of biochemical properties of amylases in germinating corn(Zea mays L.) the amylase(I), (II) and (III) from germinating corn seeds were partially purified by ammonium sulfate precipitation, DEAE-Sephadex A-50 ion exchange column chromatography and Sephadex G-100 gel filtration. The last step was effective for separation of the corn amylases to a homogeneous slate. the purified amylase(I) was identified as a kind of $\alpha$-amylase from the fact that 5％ starch solution was hydrolysed into mainly maltose and maltotetrose by it, and amylase(II) and amylase(III) were enzymes producing maltotetrose as main product. The molecular weight and specific activity of the amylase(I), (II) and (III) were determined to be 54,000 and 70.47 unit/mg, 39,000 and 62.98 unit/mg, and 51,000 and 80.39 unit/mg, respectively. It showed a tendency to increase the amylases activities in presence of Ba, Ca, Co and Fe groups, but inhibits in that of Ag, Sn, Hg and Zn groups, and amylase(I), (II) and (III) remained stable at pH 5-6 and 2$0^{\circ}C$ for 40 days in containing of 1 mM CaCl$_2$. The optimum pH and optimum temperatures were pH 6, pH 5 and pH 6 and 35$^{\circ}C$, 55$^{\circ}C$ and 55$^{\circ}C$, respectively. These results suggest that the amylase(I), (II) and (III) were different amylases.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.15 no.3
• /
• pp.276-285
• /
• 1986

This study was undertaken to investigate the characteristics of the proteolytic enzyme extracted from Neungee mushroom [Sarcodon aspratus (Berk.) S. Ito]. The enzyme was purified by using Tris-acryl CM-cellulose ion exchange, gel filtration on Ultrogel AcA 54, Hydroxy apatite column chromatography and preparative isoelectic focusing. The specific activity of the purified enzyme increased 8 times as compared with that of the crude enzyme. The enzyme was homogeneous on polyacrylamide gel electrophoresis (PAGE). The optimum pH was 10.1, indicating the enzyme to be alkaline protease and the optimum temperature was $57^{\circ}C$. The enzyme was stable at temperatures lower than $50^{\circ}C$and at pH values ranging from 4.0 to 10.8. However, the enzyme activity decreased by 26 and 65% at 60 and $65^{\circ}C$, respectively, when incubated for 30 minutes. The enzyme activity was activated by $Mn^{++}$ and inhibited by $Cu^{++}$ and $Hg^{++}$. The enzyme was consisted of monomer and its molecular weight estimated to be about 30,100 when determined by sodium dodecyl sulfate PAGE. Isoelectric point of the enzyme was determined to be 9.80.

• Korean Chemical Engineering Research
• /
• v.43 no.2
• /
• pp.187-201
• /
• 2005

Bioprocessing technologies utilizing 'biorecognition' between a solid matrix and a protein is being widely experimented as a means to replacing the conventional, solution-based technology. Frequently the matrices are chromatographic resins with specific functional groups exposed outside. Since the reactions of and interactions with the proteins occur as they are attached to the solid matrix, this 'solid-phase' processing has distinct advantages over the solution-phase technology. Solid-phase refolding of inclusion body proteins uses ion exchange resins to adsorb denaturant-dissolved inclusion body. As the denaturant is slowly removed from the micromoiety around the protein, it is refolded into a native, three-dimensional structure. Once the refolding is complete, the folded protein can be eluted by a conventional elution technique such as the salt-gradient. This concept was successfully extended to 'EBA (expanded bed adsorption)-mediated refolding,' in which the denaturant-dissolved inclusion body in whole cell homogenate is adsorbed to a Streamline resin while cell debris and other impurity proteins are removed by the EBA action. The adsorbed protein follows the same refolding steps. This solid-phase refolding process shows the potential to improve the refolding yield, reduce the number of processing steps and the processing volume and time, and thus improve the overall process economics significantly. In this paper, the experimental results of the solid-phase refolding technology applied to several biopharmaceutical proteins of various types are presented.

• The Journal of Natural Sciences
• /
• v.5 no.1
• /
• pp.67-75
• /
• 1992

In the our country, especially in Yeongil and Wolsung area, abundant authigenic zeolites are found from the tuffaceous sediments and volcanic rocks of Miocene age showing wide variation in their mineralogy and abundance from horizon to horizon. The principal zeolite species identified are clinopti-lolite. mordenite. heulandite. ferrierite, and erionite. etc. Zeolite minerals are widely used in many countries in the following applications; (a) in air separation adsorption processes; (b)as desiccants; (c)in inorganic building materials; (d)in papermaking; (e)in fertilizers; (f)as soilconditioners-this application is based upon the ability of the zeolite to ion exchange with soil nutrients; (g)in the treatment of radioactive wastes; and (h)as adsorbents for toxic gases, etc. In the present paper, using natural zeolite mordenite treated with IN hydrochloric acid or IN sodium chloride solution as column packings, separation characteristics of argon, nitrogen, carbon monoxide, and methane gases have been studied by gas chromatography. By the use of mordenite treated with hydrochloric acid solution, the tailing peak of methane showed from untreated mordenite was satisfactorily reduced, although it was difficult to separate it from carbon monoxide with a column activated at $300^{\circ}C$. Using a column activated at $350^{\circ}C$, methane could be separated from carbon monoxide easily but only carbon monoxide eluted as a bad defined peak. Mordenite treated with sodium chloride solution was generally similar to chromatograms obtained by using the untreated mordenite. Both the above chemical treatments of mordenite had little effect on the separations of argon and nitrogen. The separations and the HETP values obtained from natural zeolite mordenite treated with continuously hydrochloric acid and sodium chloride solutions were almost identical with those obtained with synthetic molecular sieve 5A zeolite. On the other hand, the efficiency of column was good in the range 20~3Oml/min of the carrier helium gas rate.

• Korean Journal of Environmental Agriculture
• /
• v.16 no.4
• /
• pp.333-340
• /
• 1997

An analytical method was developed to determine residues of haloxyfop-R and its methyl ester in soils and soybeans using gas-liquid chromatography (GLC) with electron capture detector (ECD). Soil or soybean sample was acidified and extracted with acetone. The extract was then subjected to ion-associated partition to individually separate haloxyfop-R and the neutral methyl ester. One phase containing haloxyfop-R was methylated with $BF_3$/methanol, partitioned to n-hexane and analyzed by GLC/ECD. The other phase containing the methyl ester was further purified by Florisil column chromatography prior to GLC determination. No cross contamination was found between two phases containing each of the acid and methyl ester, thus two compounds can be separately determined as the identical haloxyfop-R-methyl. Overall recoveries of haloxyfop-R from fortified samples averaged 88.2${\pm}$3.9% (n=12) and 88.3${\pm}$4.0% (n=6) for soils and soybeans respectively, and those of haloxyfop-R-methyl showed mean values of 89.2${\pm}$4.0% (n=12) and 85.6${\pm}$5.6% (n=6). Detection limits of both haloxyfop-R and its methyl esterwere 0.005㎎/㎏ and 0.01㎎/㎏ for soil and soybean samples respectively.

• The Korean Journal of Mycology
• /
• v.23 no.4 s.75
• /
• pp.332-339
• /
• 1995

Water-soluble polysaccharide (FCW) was extracted from the fruiting body of Fomitella fraxinea with neutral sodium chloride solution. The polysaccharide was further fractionated into FCW-I and FCW-II by ion exchange chromatography. The FCW-I and FCW-II were then purified by gel permeation chromatography and named as FCW-Ia and FCW-IIa, respectively. FCW-IIa showed relatively strong immuno-stimulating activity but FCW-Ia did not. By analyses of HPLC and GPC, FCW-Ia and FCW-IIa were identified to be homogeneous and their molecular weights were estimated to be about 15,000 and 8,700, respectively. FCW-Ia consisted of fucose, galactose, and mannose as main sugars and their molar ratio was 19.5 : 63.2 : 25.0. Protein was not detected in FCW-Ia. However, FCW-IIa was composed of glucose, galactose, and mannose at a molar ratio of 1.0 : 0.3 : 0.4 and contained 0.4% protein with a higher amount of glutamic acid. A small amount of uronic acid was detected in both FCW-Ia and FCW-IIa.

• Journal of Korean Society of Occupational and Environmental Hygiene
• /
• v.6 no.2
• /
• pp.292-300
• /
• 1996

Hexavalent chromium($Cr^{+6}$) compounds are considered to be particularly hazardous, primarily because of the associated risk of allergic reaction and cancer. The analytic method of hexavalent chromium such as the s-diphenylcarba-zide(DPC) method and all ether previously used methods are often made uncertain due to significant interferences from organic components. This report can provide a technique for the more rapid and simple determination of total hexavalent chromium. than other currently using methods. The s-diphenylcarbazide method proposed by the U.S. National Institute for Occupational Safety and Health has low recovery rate(15.67 - 48.20%) due to interference, iron chloride and nickel chloride. A microwave oven technique has high recovery rate(about 70%) of insoluble hexavalent chromium. For the difference of ionic charges of $Cr^{+3}$-ethylenediamine tetraacetic acid(EDTA) chelate and $CrO_4{^{-2}}$, we could detect them simultaneously by ion exchanged high performance liquid chromatography. The confirmation of $Cr^{+3}$ and $Cr^{+6}$ were checked by fraction collector and flameless atomic absorption spectrometer. We observed that the small amount of hexavalent chromium is converted to trivalent chromium due to enhancement of chromium reduction by $Fe^{+3}$ or $Ni^{+2}$. As a result of this study, on the analysis of insoluble hexavalent chromium with microwave oven was used for, it may be better and more precise analysis after pretreatment by 2% NaOH-3% $Na_2CO_3$ and then analysis UV-spectrophotometer. It should be done for various studies on insoluble hexavalent chromium on the basis work environmental monitoring so called welding, painting etc.

• Korean Journal of Food Science and Technology
• /
• v.42 no.2
• /
• pp.130-135
• /
• 2010

Picrate, enzyme-picrate and instrumental analysis methods using IC (Ion Chromatography) and HPLC (High Performance Liquid Chromatography) were compared for their effectiveness in determining cyanide in extracts of Maesil, which is classified as a harmful substance. First, the picrate method showed the shortest analysis time (about 5 hr). The color of picrate paper changed at 0.01 mg/$200\;mL\;CN^-$. However, it was difficult to detect cyanide from amygdalin of glucosides. Second, we performed a qualitative analysis for total cyanide (free cyanide and cyanide from amygdalin) by the enzyme-picrate method using $\beta$-glucosidase and a quantitative analysis by spectrophotometry. Finally, analysis of cyanide by IC and HPLC required the longest determining time (about 17 hr) as well as pretreatment for each free cyanide and amygdalin. These results suggest that enzyme-picrate is the most effective analysis method for the detection of cyanide in Maesil extracts.

• Food Science and Preservation
• /
• v.24 no.3
• /
• pp.325-333
• /
• 2017

This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents. And then, its antioxidant activity was measured using various assays including total phenolic content and protection against oxidative DNA damage, and antimicrobial activities were examined using minimum inhibiting concentration (MIC) test, and paper disc method. In addition, high-performance liquid chromatography was performed to analyze the major chemical components of ethyl acetate fraction. The G. thunbergii fraction with ethyl acetate exhibited higher antioxidant and antimicrobial activities than the other fractions. The results showed that G. thunbergii ethyl acetate fraction at $50{\mu}g/mL$ had strong DPPH and ABTS radical scavenging activities of 80.88% and 80.12%, respectively. In addition, the ethyl acetate fraction protected DNA from the oxidative damage induced by ferrous ion and hydroxyl radicals and showed high antimicrobial activity with diameter of inhibition zones ranging from 13.33 to 15.67 mm. High-performance liquid chromatography analysis revealed the major phenolic compounds of G. thunbergii to be ellagic acid and gallic acid. These results suggest that G. thunbergii might protect DNA against oxidative stress induced by reactive oxygen species and can be utilized as a natural source of antioxidant and antimicrobial agent in the food industry.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.7
• /
• pp.1043-1052
• /
• 2019

Active lipase-producing bacterium Burkholderia gladioli Bps-1 was rapidly isolated using a modified trypan blue and tetracycline, ampicillin plate. The electro-phoretically pure enzyme was obtained by purification using ethanol precipitation, ion-exchange chromatography, and gel filtration chromatography. The molecular weight was 34.6 kDa and the specific activity was determined to be 443.9 U/mg. The purified lipase showed the highest activity after hydrolysis with $p-NPC_{16}$ at a pH of 8.5 and $50^{\circ}C$, and the $K_m$, $k_{cat}$, and $k_{cat}/K_m$ values were 1.05 mM, $292.95s^{-1}$ and $279s^{-1}mM^{-1}$, respectively. The lipase was highly stable at $7.5{\leq}pH{\leq}10.0$. $K^+$ and $Na^+$ exerted activation effects on the lipase which had favorable tolerance to short-chain alcohols with its residual enzyme activity being 110% after being maintained in 30% ethanol for 1 h. The results demonstrated that the lipase produced by the strain B. gladioli Bps-1 has high enzyme activity and is an alkaline lipase. The lipase has promising chemical properties for a range of applications in the food-processing and detergent industries, and has particularly high potential for use in the manufacture of biodiesel.