• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.107-113
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• 1997

A strain producing a high amount of chitinase was isolated from soil, identified as Pseudomonas sp., and tentatively named Pseudomonas sp. YHS-A2. An extracellular chitinase of Pseudomonas sp. YHS-A2 was purified according to the procedure of ammonium sulfate saturation, affinity adsorption, Sephadex G-100 gel filtration and Phenyl-sepharose CL-4B hydrophobic interaction column chromatography. The molecular weight of the purified enzyme was estimated to be 55 kDa on SDS-PAGE was confirmed by active staining. Optimal pH and temperature of the enzyme are pH 7.0 and $50^{\circ}C$, respectively, and the enzyme is stable between pH 5.0 and 8.0 and below $50^{\circ}C$. The main products of colloidal chitin by the chitinase were N-acetyl-D-glucosamine and N,N'-diacetylchitobiose both of which were detected by HPLC analysis. The enzyme is supposed to be a random-type endochitinase which can degrade any position of ${\beta}$-l,4-linkages of chitin and chitooligosaccharides. The chitinase inhibited the growth of some phytopathogenic fungi, Fusarium oxysporum, Botrytis cineria, and Mucor rouxii and these antifungal effects were thought to be due to the characteristics of endochitinase.

• Preventive Nutrition and Food Science
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• v.1 no.1
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• pp.87-92
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• 1996

This study describes a stable and simple method for the measurement of cholesteryl ester transfer protein(CETP) activities using reconstituted HDL and LDL as substrates. Apolipoproteins (apo) A-I and -B were purified from hog plasma by a new strategy without ultracentrifugation and delipidation. a simple two-step column chromatography was administered. In the first step of phenyl-sepharose CL-4B column chro-matography, hydrophobic plasma proteins were isolated. The most hydrophobic proteins bound to the column appeared to be A-I and apo-B. Contaminat proteins were efficiently eliminated from the sample by washing the column with 0.3M NaCI containing buffer after loading the plasma on the column. Two pure proteins showing each single band on SDS-PSGE of apo A-I and apo-B were individually obtained by a subsequent gel filtration column chromatography(Sephadex G-200). This two-step purification was simple and inexpensive compared to the ultracentrifugation and/or delipidation method that are most commonly used. Reconstituted hight-density lipoproteins(HDL) and low-density lipoproteins(LDL) were prepared using the purified apo A-I and-B, respectively. When these artificially prepared HDL and LDL were used in the assays for CETP as the cholesteryl ester(CE) donor and acceptor respectively, the specific transfer of CE increased up to two fold compared to that used the native HSL and LDL.

• Preventive Nutrition and Food Science
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• v.2 no.2
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• pp.174-179
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• 1997

CMCase produced by recombinant E. coli JM109 (pCEH#4) containing CMCase gene from Cellulomonas sp. YE-5 was purified to 24.3 fold and 2.6% yield by ammoniumsulfate precipitation, DEAE-cellulose column chromatography and gel filtration on Sephadex G-100. The optimum pH and temperature for CMCase activity were pH 7.0 and 5$0^{\circ}C$. The enzyme was stable between pH 5.0 and 10.0, and up to 6$0^{\circ}C$. The molecular weight of he enzyme was estimated to be approximately 40,000 daltons by SDS-PAGE. Analysis of the amino acid composition showed that the enzyme contained many glycines and acidic amino acids. The enzyme was an endo-type CMCase and the final enzyme reaction product from hydrolysis of Cm-cellulose by the enzyme was cellobiose. {TEX}$K_{M}${/TEX} value determined with CM-cellulose was 1.28mM.

• Journal of Sericultural and Entomological Science
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• v.28 no.1
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• pp.30-36
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• 1986

Polyacrylamide gel electrophoresis and autoradiography were applied to investigate the developmental profiles of the major hemplymph proteins (MHPs) and their biosynthesis. In addition, some biochemical methods were also used to isolate and purify the MHPs. The obtained results are summarized as follows. 1. MHP-a began to appear from the 2nd day of the fourth-instar larva while MHP-b and -c were detected first on the 1st day of the fifth-instar larva. All these proteins, however, showed a drastic increase in concentration at the 2nd day of the fifth-instar larva. 2. MHP-b and -c were synthesized in fat body on early day of the fifth-instar larva, but the possibility of MHP-a synthesis in fat body was excluded. 3. MHP-b was isolated and purified by heat-treatment (6$0^{\circ}C$), gel filtration on Sephadex G-100 and column chromatography on DEAE-cellulose and CM-cellulose. Purified MHP-b showed a single band on polyacrylamide gel- and SDS-polyacrylamide gel electrophoresis.

• Korean Journal of Agricultural Science
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• v.21 no.2
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• pp.122-132
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• 1994

Bacteriostatic activity of secretory immumoglobulin A (SIgA) in human and bovine colostrums on enterotoxigenic type Salmonella was tested in the tissue culture medium. SIgA was used in $0.1{\sim}5.0mg/m{\ell}$ concentration with or without the addition of egg lysozyme tested for theirs bacteriostatic activites. 1. Bovine SIgA rich fraction with a large amount of $IgG_1$-dimer could be prepared from bovine colostrum of Holstein cows by anion exchange chromatography using DEAE-Sephadex A-50 and gel filtration on Sephadex G-200 and Sepharose 6B. 2. Human SIgA appeared to be the most bacteriostatic effect for all varieties of Salmonella in a range of $0.5{\sim}1.0mg/m{\ell}$ Bovine SIgA showed a marked bacteriostatic effect increased by increasing the concentration. Bovine IgG had not show bacteriostatic effect against both enterotoxigenic type Salmonella. Egg lysozyme as well as bovine SIgA also showed a marked bacteriostatic effect increased by increasing the concentration. 3. When the growth inhibition of human SIgA was tested by adding egg lysozyme with time interval, egg lysozyme showed bacteriostatic effect as compared with control. But human SIgA and adding with lysozyme showed slight the bacteriostatic effect. 4. When the growth inhibition of bovine SIgA was tested by adding egg lysozyme with time interval, all treatment against S. enteritidis showed bacteriostatic effect as compared with control In the case of S. typhinwrium, egg lysozyme showed a marked bacteriostatic effect as compared with control.

• Korean Journal of Food Science and Technology
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• v.9 no.3
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• pp.211-219
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• 1977

Dehulled and defatted Korean cottonseed flour was extracted with alkaline solution for 30 minutes and had precipitated the crude portein by adjusting pH $1{\sim}12$. The general composition and the amino acid composition of cottonseed protein were analyzed. Crude protein was purified with sephadex G-100 and G-200, and its component had been identified by disc electrophoresis. Toxic gossypol was removed by n-hexane, acetone and other solvents. The results were as follows. (1) pH 5, pH 7 and pH 4 were the best condition of precipitation of curde protein at single, two step and water extraction, respectively. (2) The cottonseed flour which was dehulled and defatted, contained 61.3% of crude protein. (3) The protein which was isolated from cottonseed flour, contained 20% of glutamic acid, and comparatively high levels of essential amino acids. (4) Dehulled cottonseed flour contained 0.97% of total gossypol and could be romoved 70% of total gossypol by extraction with n-hexane. (5) 10-13 bands of water soluble protein were found in disc electrophoresis, and 10-12 bands in protein were isolated by single and two step procedures. (6) The cottonseed protein could be purified by sephadex G-100 and G-200. (7) 10-20% of gossypol-free cottonseed fluor could be used for animal and human comsumption.

• Korean journal of applied entomology
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• v.12 no.3
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• pp.93-107
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• 1973

Garlic (Allium sativum L.) is an important vegetable crop for the Korean people and has long been cultivated extensively in Korea. More recently it has gained importance as a source of certain pharmaceuticals. This additional use has also contributed to the increasing demand for Korean garlic. Garlic has been propagated vegetatively for a long time without control measures against virus diseases. As a result it is presumed that most of the garlic varieties in Korea may have degenerated. The production of virus-free plants offers the most feasible way to control the virus diseases of garlic. However, little is known about garlic viruses both domestically and in foreign countries. More basic information regarding garlic viruses is needed before a sound approach to the control of these diseases can be developed. Currently garlic mosaic disease is most prevalent in plantings throughout Korea and is considered to be the most important disease of garlic in Korea. Because of this importance, studies were initiated to isolate and characterize the garlic mosaic virus. Symptom expression in test plants, physical properties, purification, serological reaction and morphological characteristics of the garlic mosaic virus were determined. Results of these studies are summarized as follows. 1. Surveys made throughout the important garlic growing areas in Korea during 1970-1972 revealed that most of the garlic plants were heavily infected with mosaic disease. 2. A strain of garlic mosaic virus was obtained from infected garlic leaves and transmitted mechanically to Chenopodium amaranticolor by single lesion isolation technique. 3. The symptom expression of this garlic mosaic virus isolate was examined on 26 species of test plants. Among these, Chenopodium amaranticolor, C. quince, C. album and C. koreanse expressed chlorotic local lesions on inoculated leaves 11-12 days after mechanical inoculation with infective sap. The remaining 22 species showed no symptoms and no virus was recovered from them whet back-inoculated to C. amaranticolor. 4. Among the four species of Chtnopodium mentioned above, C. amaranticolor and C. quinoa appear to be the most suitable local lesion test plants for garlic mosaic virus. 5. Cloves and top·sets originating from mosaic infected garlic plants were $100\%$ infected with the same virus. Consequently the garlic mosaic virus is successively transmitted through infected cloves and top-sets. 6. Garlic mosaic virus was mechanically transmitted to C, amaranticolor when inoculations were made with infective sap of cloves and top-sets. 7. Physical properties of the garlic mosaic virus as determined by inoculation onto C. amaranticolor were as follows. Thermal inactivation point: $65-70^{\circ}C$, Dilution end poiut: $10^-2-10^-3$, Aging in vitro: 2 days. 8. Electron microscopic examination of the garlic mosaic virus revealed long rod shaped particles measuring 1200-1250mu. 9. Garlic mosaic virus was purified from leaf materials of C. amaranticolor by using two cycles of differential centrifugation followed by Sephadex gel filtration. 10. Garlic mosaic virus was successfully detected from infected garlic cloves and top-sets by a serological microprecipitin test. 11 Serological tests of 150 garlic cloves and 30 top-sets collected randomly from seperated plants throughout five different garlic growing regions in Korea revealed $100\%$ infection with garlic mosaic virus. Accordingly it is concluded that most of the garlic cloves and top-sets now being used for propagation in Korea are carriers of the garlic mosaic virus. 12. Serological studies revealed that the garlic mosaic virus is not related with potato viruses X, Y, S and M. 13. Because of the difficulty in securing mosaic virus-free garlic plants, direct inoculation with isolated virus to the garlic plants was not accomplished. Results of the present study, however, indicate that the virus isolate used here is the causal virus of the garlic mosaic disease in Korea.

• Clinical and Experimental Reproductive Medicine
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• v.10 no.2
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• pp.1-11
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• 1983

${\beta}$-Glucuronidase from bull seminal plasma was partially purified by $(NH_4)_2SO_4$fractionation, two successive DEAE-cellulose columns, isoelectric focusing (pH 4 to 6) and Gel filtration on Sephadex G-200. Only one form of ${\beta}$-glucuronidase was obtained by isoelectric focusing at pH 5.13. Highly purified ${\beta}$-glucuronidase had specific activity of 34 units/mg protein and showed one major and some minor contaminants by disc gelk electrophoresis. The enzyme showed maximum activity at pH 5.2 and at $48^{\circ}C$. The enzyme was completely inhibited by 1,4 saccharo-${\alpha}$-lactone (5 mM). Albumin and 0.15 M NaCl increased the ${\beta}$-glucuronidase activity. Km of ${\beta}$-glucuronidase using phenolphthalein mono-${\beta}$-glucuronic acid as substrate was 2.9 mM and Vmax was $0.8{\mu}$mole/min. The enzyme appeared to be a glycoprotein by its binding to concanvalin·A. Rabbit and human sperm-acrosomal extracts and seminal plasma showed high ${\beta}$-glucuronidase activity.

• Korean journal of applied entomology
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• v.18 no.2 s.39
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• pp.89-100
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• 1979

[ $\lceil$ ]한국산 명나방과(Pyralidae)에 관하여는 이미 필자(1976)에 의하여 언급된바와 같이 1889년 영국인 곤충학자 J.H. Leech가 처음으로 9종의 신종을 발표한데 이어 현재까지 300여종이 기록되어 왔다. 그러나 이들의 기록이 여러종류의 발표문에 산재해 있는 상태로 체계적인 정리가 이루어진바 없으며 학명뿐아니라 우리말 이름조차 갖가지로 쓰여지고 있어 곤충명 사용에 혼란이 야기되고 있는 실정이다. 이에 필자는 현재까지 발표되었던 종들을 조사하여 최근의 분류체계에 따라 학명 및 그들의 동의어(Synonym)를 정리함과 아울러 우리말 이름도 먼저 명명된 이름이나 또는 현재 많이 사용되고 있는 이름을 택하여 통일화하고 ( )안에 그 이명을 표시하였다. Evergestiinae(새들명나방아과, 신칭)은 현재까지는 주로 Pyraustinae(들명나방아과)에 포함되는 족(Tribe)으로 취급되어 왔으나 최근의 분류체계에 따라 독립된 아과로 취급하였다. 이들에 대해서는 Leech에 이어 일본인 Shibuya(1927년에 의하여 13종, 박세욱등 (1958, 1969)에 의하여 27종이 새로 기록되는등 1968년에 출판된 한국 동물명집 곤충편에는 94종이 수록되었다. 최근 필자(1976)에 의하여 우리나라 미기록종으로 발표된 24종과 기타 국내외의 문헌을 통한 기록의 근거를 기초로 작성된 이 목록에는 Evergestiinae에 1속 3종, Pyraustinae에 53속 123종이 포함되었으며 학명이 불분명하거나 채집의 근거가 불확실한 4종은 후미에 별도로 기재하였다. 분포란의 괄호안에 명시된 채집지는 필자에 의하여 채집 확인된 종들로서 그들의 표본은 농업기술연구소 곤충연구담당관실 표본실에 소장되어 있다. 다른 아과에 관하여는 정리가 완료되는대로 추후 발표할 예정이다.nemella hengsungica, Aphelenchoides besseyi 등으로 2아목 7과 11종이었다.는 내연가공한 옷생산에 사용되지 않는 것이 타당하겠다. 주아의 즙액에 의해서도 C. amaranticolor에 기계적 전염이 되었다. 7. C. amaranticolor 상에 계통분리된 마늘 모자이크 바이러스의 내열성은 $65^{\circ}-70^{\circ}C$, 희석한계는 $10^{-}2-10^{-3}$, 그리고 보존한계는 2 일이었다. 8. 마늘 모자이크 바이러스의 순화는 2회의 분획원심과 Sephadex gel filtration에 의해서 가능했다. 9. 전자현미경하에서 관찰한 마늘 모자이크 바이러스는 길이 1200-1225mu 폭 10-12mu의 사상이었다. 10. 혈청학적 미량침강 반응법에 의해 마늘잎에서뿐만 아니라 인편과 주아에서도 마늘 모자이크 바이러스의 검정이 가능했다. 11. 우리나라 5개 지방에서 수집한 마늘 종구 150개와 주아 30개에 대해 혈청학적방법으로 마늘 모자이크 바이러스의 감염률을 조사한 결과 $100\%$의 감염률을 보였다. 12. 마늘 모자이크 바이러스와 크기가 근사한 Potato Virus X. Potato virus Y, Potato virus S, Potato virus M 등과의 혈청학적 유연관계를 조사한 바, 마늘 모자이크 바이러스는 이들과 구별되는 다른 바이러스라고 생각된다. 13. 마늘의 모자이크 감염주에서 단일계통으로 분리하여 본 실험에 사용한 바이러스는 마늘의 바이러스 무감염주를 얻을 수가 없기 때문에 직접 마늘잎에 접종해서 모자이크톤의 병원이라는 것을 확인할 수 없었지만, 검정식물상의 반응, 혈청학적반응, 전자현미경적 관찰등의 간접적인 조사 결과로 미루어 미인록의 마늘모자이크 바이러스라고 생각된다

• Biomolecules & Therapeutics
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• v.1 no.2
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• pp.188-195
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• 1993

In order to understand the machanism of action and regulation of ${\beta}$-adrenergic receptor in terms of molecular level, the purification of receptor protein has a fundamental importance. Moreover, species differences among avian, amphibian and mammalian ${\beta}$-adrenergic receptors make it more important to purify mammalian ${\beta}$-adrenergic receptor. Because ${\beta}$-adrenergic receptor is an integral membrane protein, it must be solubilized from the membrane for the purification. The purpose of the present study was to solubilize and characterize the mammalian $\beta$-adrenergic receptor from guinea pig lung in quantities by more efficient and practical method eventually to purify receptor. Guinea pig lung membrane preparation was solubilized by sequential treatment of buffers containing low and high concentration of digitonin which are 0.2 and 1.2% respectively. About 50% of the total receptor pool was released by this double extraction procedure. The $\beta$-adrenoceptors in the digitonin extract were identified using the ${\beta}$-adrenergic antagonist, (-)-[$^3H$]-dihydroalprenolol ([$^3H$]DHA). The solubilized receptor retained all of the essential characteristics of membrane-bound receptor, namely saturability; stereoselectivity; high affinity to ${\beta}$-adrenergic drugs. For the measurement of soluble receptor activity, Sephadex G-50 chromatography method has been widely used. Inspite of its accuracy and wide acceptance, this technique employed troublesome column work which required long time to assay the activity of receptor. We employed another methods to measure receptor activity. When using 0.5% polyethylenimine pretreated GF/B glass fiber filter, filtration technique could be used to measure soluble receptor activity. This technique enabled us to reduce the total amount of time to assay by a factor of 4 as well as to detect soluble receptor. In the present study, we could establish more efficient and practical solubilization method of mammalian $\beta$-adrenergic receptor. The rapidity and high yield of this solubilization scheme, together with the favorable recovery of the receptor activity, are significant steps toward the ultimate purification of the mammalian $\beta$-adrenergic receptor. The result of this study together with more convenient purification method could provide large amount of purified receptor with ease for various research purposes.