• Journal of Microbiology
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• 제33권2호
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• pp.109-114
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• 1995

Alkalophilic Cephalosporium sp. RYM-202 produced multiple xylanases extracellularly. One of these xylanases was purified to electrophoretical homogeneity by chromatography with DEAE-Sephadex A-50, Sephacryl S-200 HR and Superose 12 HR. The purified xylanase differed from most other microbial xylanases in that it had low-molecular weight and acidic isoelectric point. The molecular weight of the xylanase in that it had low-molecular weight and acidic isoelectric point. The molecular weight of the xylanase was 23 kDa by SDS-polyacrylamide electrophoresis and 24 kDa by gel permeation chromatography, and the isoelectric point was 4.3. The xylanase had the highest activity permentation chromatography, and the isoelectric point was 4.3. The xylanase had the highest activity permeation chromatography, and the isoelectric point was 4.3. The xylanase had the highest activity at pH 8.0 and 50 .deg.C. It was stable over a wide range of pH and retained more than 80% of its original activity after 24 h of incubation even at pH 12. The Km values of this enzyme on birchwood xylan and oat spelts xylan were 2.33 and 3.45 mg/ml, respectively. The complete inhibition of the enzyme of n-bromosuccinimide suggests the involvement of tryptophan in the active site. The sylanase lacked activity towards crystalline cellulose and carboxymethyl cellulose.

• 환경생물
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• 제29권4호
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• pp.305-311
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• 2011

$Lactobacillus$ $jensenii$ YW-33이 생산하는 생물응집제의 활성본체를 규명하고자 응집물질을 분리, 정제하고 그 특성을 검토하고자 하였다. 생물응집제의 활성 본체를 조사하기 위하여 배양액을 원심분리하여 균체 자체와 균체를 제거한 상등액을 비교한 결과 균체를 제거한 상등액이 86%로 높은 응집활성을 나타냈다. 또한 pronase 처리와 periodate 산화를 행한 결과 pronase로 처리한 시료는 무처리군과 비교하여 차이가 없었던 반면 periodate로 산화시킨 시료는 응집활성이 크게 감소함에 따라 다당에 기인되는 것으로 추정되었다. 균체가 제거된 상등액을 감압, 농축하여 EtOH 60~80%의 농도로 분획한 결과 LJ-80을 얻었다. 분리된 LJ-80을 DEAE-Toyopearl 650C chromatography, Sepharose CL-6B chromatography을 이용하여 최종 정제된 LJ-80-II-1을 분리하였으며 HPLC 분석에 의해 순도를 확인했다. 정제된 LJ-80-II-1의 분자량은 약 800,000 이상이며 총당이 98.4%, 단백질이 0.6%으로 mannose : galactose : glucose가 1.61 : 0.25 : 1.00의 molecule ratio을 가지고 있었다.

• 한국식품과학회지
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• 제31권3호
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• pp.848-854
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• 1999

전통된장으로부터 안지오텐신전환효소(angiotensin converting enzyme; ACE)를 저해하는 물질을 추출하여 그 구조를 밝혀냈다. ACE 저해물질을 열수추출한 다음 gel permeation chromatography (GPC)를 통하여 ACE 저해작용이 큰 두 개의 큰 획분을 받았다. 앞획분은 90%와 70%의 ACE 저해효과를 나타내었으나 단일물질로 분리되지 않아 계속하여 역상 HPLC를 통하여 순수 분리를 하였다. 그러나 앞획분은 순수분리되지 않아 결국 2차원 전기영동/TLC를 통하여 분리한 결과 분자량이 759.63인 아미노기를 갖고 있는 비펩타이드 물질임이 밝혀졌다. 뒷획분은 다른 조건의 HPLC(reverse column과 $NH_2$, column)를 이용하여 순수분리에 성공하였다. 이중 ACE 저해효과가 큰 물질은 분자량 271.33인 dipeptide인 arginine-proline임을 밝혀냈다. 이물질의 ACE $IC_{50}$은 $92\;{\mu}M$이었다. 본 연구 결과는 대부분 ACE 저해물질이 3개 내지 7개 등의 긴 펩타이드임을 감안할 때, 짧은 dipeptide로 ACE 저해펩타이드가 한국의 전통된장에서 생산할 수 있음을 보여주고 있다.

• 한국식품과학회지
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• 제21권1호
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• pp.164-172
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• 1989

Streptococcus thermophilus 510의 ${\beta}-galactosidase$는 유당을 가수분해 하는 동시에 galactose전이반응을 일으켜 세 종류의 galactooligosaccharides를 형성 하였다. Oligosaccharide의 최대 생성 조건은 40% 유당 용액에 효소 50 ONPG units/ml를 첨가한 혼합액을 $37^{\circ}C$에서 4시간 반응시켰을 때 이며, 이 조건에서 유당이 약 94% 가수분해 되었고 생성된 oligosaccharides의 양은 총 당의 약 30% 이었다. 생성된 총 oligosaccharides의 69%는 $6-o-{\beta}-D-galactopyranosyl-D-glucose(allolactose)$, 23%는 $6-o-{\beta}-D-galactopyranosyl-D-galactose(isogalactobiose)$ 이었으며, 형성된 galactooligosaccharides을 Bio-Gel P-2 gel permeation chromatography 방법에 의해서 순수하게 분리 정제하여 구조를 조사하여 본 결과 glucose, galactose, allolactose, 그리고 isogalactobiose 임을 확인할 수 있었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제6권2호
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• pp.151-155
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• 2003

Using gel filtration chromatography, high molecular silk fibroin with high purity was obtained and silk flbroin microsphere particles (SFMP) could be simply made by spray dryer method. Also, some of the physicochemical properties of SFMP and morphology were investigated. The average molecular weight of pure silk fibroin protein dissolved in calcium chloride is about 61,500g/㏖ as measured by gel permeation chromatography. SFMP was spherical in shape, and particles, sized average of 2 ${\pm}$ 10 ${\mu}$, were observed by SEM and particle analyzer, respectively. Obtaining microspheres particles by spray dryer method accelerated the transition from the random coil to the $\beta$-sheet structure during spray dryer treatment. It was identified by the basic fourier transform infrared spectroscopy of SFMP. The swelling ratio of SFMP is majorly dependent on the pH of the solution, not on the occurred gelation. The characteristic structure, which might be applicable to immobilization of drugs is superior to other matrix materials for the use of biomaterials with skin affinity.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.957-963
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• 2001

Bacillus subtilis YL-1004 was isolated from soil for the development of agents to control dental caries. This strain produced an extracellular lytic enzyme that hydrolyzed the Streptococcus mutans cell wall. The lytic enzyme was purified to homogeneity by affinity chromatography and gel permeation chromatography to give a single band on SDS-PAGE and non-denaturing polyacrylamide gel electrophoresis. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography to be 38 kDa and the PI to be 4.3 from isoelectric focusing. Sirty $\%$ of its lytic activity remained after incubation at $50^{\circ}C$ for 30 min, and its optimal temperature was $37^{\circ}C$ . The enzyme showed its highest activity at pH 8.0 and was stable at pHs ranging from 4.0 to 9.0. Treatment with several modifiers showed that a cysteine residue was involved in the active site of the enzyme. This lytic enzyme from Bacillus subtilis YL-1004 exhibited specificity towards Streptococci and also showed autolytic activity on Bacillus subtilis YL-1004.

• 한국식품영양학회지
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• 제34권1호
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• pp.26-35
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• 2021

Myrosinases (thioglucosidases) catalyze the hydrolysis of a class of compounds called glucosinolates, of which the aglycones show various biological functions. It is often necessary to minimize the loss of myrosinase activity during thermal processing of cruciferous vegetables. Myrosinase was isolated from a popular spice, white mustard (Sinapis alba), and its thermal inactivation kinetics was investigated. The enzyme was extracted from white mustard seeds and purified by a sequential processes of ammonium sulfate fractionation, Concanavalin A-Sepharose column chromatography, and gel permeation chromatography. At least three isozymes were revealed by Concanavalin A-Sepharose column chromatography. The purity of the major myrosinase was examined by native polyacrylamide gel electrophoresis and on-gel activity staining with methyl red. The molecular weight of the major enzyme was estimated to be 171 kDa. When the consecutive step model was used for the thermal inactivation of the major myrosinase, its inactivation energy was 44.388 kJ/mol for the early stage of destruction and 32.019 kJ/mol for the late stage of destruction. When the distinct two enzymes model was used, the inactivation energy was 77.772 kJ/mol for the labile enzyme and 95.145 kJ/mol for the stable enzyme. The thermal inactivation energies lie within energy range causing nutrient destruction on heating.

• Journal of the Korean Wood Science and Technology
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• 제42권6호
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• pp.740-746
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• 2014

꾸지뽕나무 미숙과를 50% 발효주정으로 추출하고, 농축한 추출용액을 n-hexane, chloroform, ethyl acetate, butanol로 순차적으로 분액하여 수용성 조추출물을 얻었다. 수용성 조추출물을 분취용 겔 투과 크로마토그래피(Gel Permeation Chromatography, GPC)를 실시하여 다당류(F1)를 분리하였다. F1을 가수분해하여 단당 성분 분석을 실시한 결과 glucose, galactose, arabinose, xylose 등 4가지로 구성되어 있었으며, 그 조성비는 6.1 : 1.8 : 3.1 : 1.0이었다. F1에 acetyl 기의 치환여부와 uronic acid의 존재 유무를 확인하기 위해 FT-IR로 측정하고 염산 가수분해 한 다음 HPLC 분석을 실시한 결과, 두 가지 모두 없는 것으로 분석되었다. 결합 위치 확인을 위해 permethylation, 산 가수분해, 환원, silylation을 순차적으로 실시하여 얻은 partially methylated alditol silylate (PMAS)들을 GC-MS 분석을 실시한 결과, 1,5-di-O-trimethylsilyl-2,3,4-tri-O-methyl xylose, 1,5-di-O-trimethylsily-2,3,4-tri-O-methyl arabinose, 1,4,5-tri-O-trimethylsily-2,3-di-O-methy arabinose, 1,3,5-tri-O-trimethylsily-2,4,6-tri-O-methyl glucose, 1,4,5-tri-O-trimethylsily-2,3,6-tri-O-methyl galactose, 1,4,5-tri-O-trimethylsily-2,3,6-tri-O-methyl glucose, 1,3,5,6-tetra-O-trimethylsily-2,4-di-O-methyl galactose, 1,3,5,6-tetra-O-trimethylsily-2,4-di-O-methyl glucose로 구성되어 있었고 그 조성비는 1.1 : 1.0 : 4.9 : 7.5 : 3.0 : 3.1 : 1.4 : 1.5였다.

• Preventive Nutrition and Food Science
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• 제5권3호
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• pp.148-152
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• 2000

Mussel adhesive protein (MAP) was extracted from Korean Mytilus edulis and then partially purified using Sephacryl S-300 gel permeation chromatography and reversed-phase high performance liquid chromatography. As an indicator of adhesiveness, is 3,4-dihydroxyphenylalanine (DOPA) content was determined. Its DOPA/protein ratio of 0.19 was higher than those of other reports, indicating a good adhesive. The partially purified MAP was confirmed by acid-urea polyacrylamide gel electrophoresis using cetylpiridinium bromide as a cationic detergent. Sea mussel hydrolysates were prepared using three commercial proteases to provide value-added functional materials and their angiotensin converting enzyme (ACE) inhibitory activities were determined. Among hydrolysates of sea mussel, Protamex was the best and further purification would improved ACE inhibitory activity.

• 미생물학회지
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• 제25권2호
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• pp.148-156
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• 1987

Extracellular laccase (E.C. 1.10.3.2) from the culture filtrate of Pleurotus ostreatus was purified by ammonium sulfate precipctation, protamine sulfate precipitation, DEAE-Sephadex A-50 ion exchange chromatography and Sephadex G-100 gel permeation chromatography. The molecular weight of the enzyme was estimated by SDS-polyacrylamide gel electrophoresis to be 58,000 and the isoelectric point was 3.75. The optimum temperature for the enzyme was about $45^{\circ}C$ and the optimum pH was 6.5. The enzyme was found to be stable at temperature below $35^{\circ}C$ and rapidly inactivated at higher temperatures. Km values for ferulic acid, vanillic acid, dihydroxyphenylalanine (DOPA) were 48.6.$\mu$M, 0.52mM, and 2.73mM, respectively, which indicates that the enzyme has much higher affinity towards ferulic acid. The reaction products of the enzyme were separated by TLC and HPLC.