폐단백자원에 이용하기 위한 미생물 Protease 의 특성

천성숙,조영제,성태수,손준호,최청 ( Sung Sook Chun,Young Je Cho,Tae Soo Sung,Jun Ho Son,Cheong Choi ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.1

To extract insoluble proteins and to improve functional properties of abolished proteins, a protease producing Aspergillus sp. MS-18 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of protease reached to the maximum when the wheat brae medium containing, 3% arabinose, 0.5% polypepton, 0.1% (NH₄)₂SO₄ and 0.2% magnesium chloride was cultured for 3 days. Protease was purified 16.9 folds after ion exchange chromatography and gel filtration and the specific activity was 340.4 unit/㎎. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of protease was estimated to be 30,000. Crystalization form of purified protease was a stick shape with rounding edges. The optimum pH and temperature for the protease activity were 9.0 and 60℃, respectively. The enzyme was stable in pH 7.0-12.0 at 50℃. The activity of purified enzyme was inhibited by Hg^(2+), Zn^(2+) and Pb^(2+), whereas it was activited by Na^+, Mg^(2+) and Mn^(2+). The activity of the protease was inhibited by the treatment with ethylenediaminetetraacetic acid and phenylmethane sulfonyl fluoride. The result suggests that the purified enzyme is a serine protease with metal ion at active site. Km and Vmax of purified protease were 29.33 μmole/L and 5.13 ㎍/min, respectively.

Protease 처리에 의한 폐단백자원의 단백질 용출 및 기능성 변화

천성숙,조영제,손규목,최희진,최청 ( Sung Sook Chun,Young Je Cho,Gyu Mok Son,Heui Jin Choi,Cheong Choi ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.1

To improve extraction of insoluble proteins and functional properties of abolished proteins by protease. It was found that the optimum pH, optimum temperature, optimum treatment time and optimum unit of enzyme far extraction of protein were pH 9.0, 60℃, 8 hrs, 40 units. The foaming capacity and foaming satbility of sesame meal protein after treatment of enzyme were especially higher than control. The emulsion capacity and emulsion satbility of sesame meal protein were higher than control. Coil absorption as well as water absorption capacities of sesame meal protein were higher than control.

포스터 발표논문 초록 ( 식품 / 기타 분야 Poster 초록 ) : Protease 처리에 의한 폐단백자원의 단백질 용출 및 기능성 변화

천성숙,죄영제,손규목,손준호,최희진,최청 한국응용생명화학회(구 한국농화학회) 1997 한국응용생명화학회 학술발표회 Vol.1997 No.2

Aspergillus속 균주가 생산하는 Phytase의 분리 정제 및 특성

천성숙(Sung-Sook Chun),조영제(Young-Je Cho),차원섭(Woen-Suep Cha),이희덕(Hee-Duck Lee),이선호(Sun-Ho Lee),최청(Chung Choi) 한국식품영양과학회 1998 한국식품영양과학회지 Vol.27 No.1

단백질과 결합하여 단백질의 용해도를 저하시키고, 무기질의 흡수를 저해하는 phytic acid의 효소에 의한 분해, 제거를 목적으로 phytase를 생산하는 Aspergillus sp. SM-15 균주를 토양으로 부터 분리하고 이 균주가 생산하는 효소를 정제하여 특성을 규명하였다. Phytase생산조건을 밀기울배지에 1.0% mannose, 2% yeast extract, 0.5% (NH₄)₂HPO₄, 0.2% calcium chloride를 첨가하여 4일 배양시 최대 활성을 나타내었다. 효소는 ion exchange chromatography, gel filtration 등으로 약 17.1배 정제하였고 효소의 비활성역가는 244.32unit/㎎이었다. 정제효소는 polyacrylamide gel 전기영동상 단일밴드로 나타났으며, 분자량은 46,000 전도로 추정되었다. 정제효소의 특성은 최적 pH와 온도는 각각 5.0, 50℃였으며, 안정범위는 pH 4.5~5.5까지 였다. 금속이온 중 Fe^(2+), Pb^(2+) 등에 의해 활성이 증대되었으나, Hg^(2+)에 의해 효소활성이 저해되었고, 저해제 중 iodine에 의해서 활성저해가 관찰되어 active site에 histidine 잔기가 존재하는 것으로 추정되었으며, 정제효소의 Km과 Vmax는 각각 37.037mM/L, 159.87μ㏖/min이었다. To extract insoluble proteins and to improve funtional properties of abolished proteins, an phytase producing Aspergillus sp. SM-15 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. Phytase production reached to maximum when the wheat bran medium containing 1% mannose, 1% yeast extract, 1% (NH₄)₂HPO₄ and 0.2% calcium chloride was cultured for 4 days. Phytase was purified 17.1 fold and specific activity was 244.32unit/㎎ by a sequencial precess of ammonium sulfate fraction, ion exchange chromatography and gel filtration. Purified enzyme was confirmed as a single band by the polyacrylamide gel electro-phoresis. The molecular weight of phytase was estimated to be 46,000. The optimum pH and temperature for the phytase activity were 5.5 and 50℃. The enzyme is stable in pH 4.5~5.5, 60℃. The activity of purified enzyme was inhibited by Hg^(2+) whereas activited by Pb^(2+) and Fe^(2+). The activity of phytase was inhibited by the treatment with iodine. The result indicate the possible involvement of histidine at active site. Km and Vmax of the purified phytase were 37.037mM/L and 159.87μ㏖/min, respectively.

Phytase 처리에 의한 폐단백자원의 단백질 용출 및 기능성 변화

천성숙(Sung-Sook Chun),조영제(Young-Je Cho),김영활(Young-Hwal Kim),우희섭(Hi-Seob Woo),최청(Chung Choi) 한국식품영양과학회 1998 한국식품영양과학회지 Vol.27 No.1

폐단백질을 활용하는 방도의 하나로 폐단백질자원으로부터 토양에서 분리한 Aspergillus sp. 균주가 생산하는 phytase를 이용하여 불용성 단백질의 분리 효율성을 높였으며 추출 단백질의 기능성을 살펴보았다. 참깨박에 함유되어 있는 불용성 형태의 단백질을 가용성 형태의 단백질로 용출시키기 위한 최적 pH, 최적 온도, 최적 처리시간과 최적 첨가효소량은 4.0~5.0, 50℃, 8~10시간, 120unit였고, 효소처리된 참깨박은 phytase 처리의 경우 대조구에 비해 기포형성력은 크게 증가하지 않았으나 기포안정성은 다소 증가하였다. 참깨박 단백질의 유화력은 다소의 유화력과 안정성의 증가가 관찰되었고, 유지흡착력과 수분흡착력은 대조구에 비해서 높은 값을 나타내었다. This study was performed to improve extraction of insoluble proteins and to evaluate funtional properties of abolished proteins by the phytase produced by Asporgillus sp. The optimum pH, temperature, treatment time and unit of the enzyme for extraction of protein were pH 4.0~5.0, 50℃, 8~10hrs and 120 units. The foaming capacity and foaming stability of sesame meal protein after enzyme treatment were virtually unchanged as compared to control. The emulsion capacity and emulsion stability of sesame meal protein was higher than control. Oil absorption as well as water absorption capacities of sesame meal protein were higher than control.

Phytase와 Protease 혼합처리에 의한 참깨박 단백질 용출 및 기능성 변화

천성숙(Sung-Sook Chun),조영제(Young-Je Cho),조국영(Kuk-Young Cho),최청(Cheong Choi) 한국식품과학회 1998 한국식품과학회지 Vol.30 No.4

Protease 처리가 누에번데기 단백질 추출 및기능성에 미치는 영향

권효정,천성숙,조영제,차원섭,이경환,김정환 한국응용생명화학회 2006 Journal of Applied Biological Chemistry (J. Appl. Vol.49 No.4

추출용매 비에 따른 백부자(Aconiti koreani Rhizoma) 추출물의항균효과 및 항산화효과

윤소정,천성숙,조영제,김정환,이경환,권효정 한국응용생명화학회 2005 Journal of Applied Biological Chemistry (J. Appl. Vol.48 No.3

For the purpose of developing natural antioxidant, the antioxidative activity and antimicrobial of phenolics isolated from Baek-bu-ja (Aconiti koreani Rhizoma) were determined. Optimum extracting condition for phenolics were water extracts. HPLC analysis showed that the four major phenolic metabolites were rosemarinic, protocatechuic, caffeic and chlorogenic acids. The water extracts of Baekbuja did not have antimicrobial activity against H. pylori; however, the ethanol extracts revealed higher antimicrobial activity. Electron donation ability on DPPH of Baekbuja ethanol extract was 20% higher than other ethanol extracts. The 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid radical decolorization (ABTS) and antioxidant protection factor (PF) were determined with extracts from Aconiti koreani Rhizoma. 94% inhibition and 1.14 PF were shown on ABTS and antioxidant protection factor with 60% ethanol extracts. Also, TBARS (thiobarbituric acid reactive substances) showed 0.19 µM in the control and 0.07 µM in the 80% ethanol extracts. The result suggests that Baekbuja extract may be useful as potential sources of anti Helicobacter pylori, antioxidant 백부자(Aconiti koreani Rhizoma)를 이용한 항암효과와 노화억제를 위한 기능성 식품 소재로 개발하기 위하여 항균 및 항산화효과를 조사하였다. 백부자 추출물의 phenol성 물질의 추출수율은 열수추출물이 8.45±0.27 mg/g이었으며, 에탄올추출물은 20, 40, 60, 80, 100% 농도별로 각각 6.40±0.10 mg/g,6.53±0.26 mg/g, 6.96±0.25 mg/g, 2.44±0.04 mg/g, 0.31±0.17mg/g으로 열수추출물의 phenol 함량이 다소 높게 나타났다. 항산화 효과가 높다고 알려진 phenol 성분의 함량을 HPLC 분석결과 항산화 효과가 높은 protocatecuic acid, chlorogenic acid, rosemarinic acid의 함량이 높게 나타났다. H. pylori에 대한 항균효과는 에탄올추출물에서만 관찰되었다.. ABTS radical decolorization과 antioxidant protection factor(PF)를 살펴 본 결과 ABTS는 60% 에탄올추출물에서 94.25±0.68%로 가장 높은 저해율을 나타내었고, PF 역시 60% 에탄올추출물에서 1.14±0.05로 비교적 높은 protection factor을 나타내었다. DPPH에 대한 전자공여능은 20% 에탄올추출물에서 93.69±0.19%의 가장 높은 활성을 나타내었으나, 40%, 60%, 80%간의 유의적인 차이는 없었고, 열수추출물에서는 60.10±1.63%의 낮은 활성을 나타내었다. 활성산소 중 지방산화를 일으키는데 중요한 역할을 하는 hydroxyl radical에 대한 각 추출물들의 영향은 80% 에탄올추출물이 다른 농도의 추출물들에 비하여 낮은 TBARS값을 나타내었다. 이상의 결과로 볼 때 백부자로부터 추출한 페놀성 물질은 60%의 ethanol로 추출한 추출물이 천연항산화 물질로 활용이 가능한 기능성 식품의 소재로의 개발이 가능할 것으로 사료된다.

Bacillus속이 생산하는 Cyclodextrin Glucanotransferase에 의한 Stevioside로의 당전이반응

조영제,천성숙 한국응용생명화학회 2004 Journal of Applied Biological Chemistry (J. Appl. Vol.47 No.1

Cyclodextrin glucanotransferase (CGTase) of Bacillus sp. isolated from soil was purified and its enzymological characteristics were investigated. It was found that the production of CGTase reached to the maximum when the strain was cultured in the broth containing 0.1% albumin, 2% NH4Cl, 2% soluble starch and 0.2% KH2PO4 for 72 hrs at 37oC. The purity of CGTase was increased by 9.7 folds through purification procedures by the following column chromatography DEAE-cellulose ion exchange chromatography and Sephadex G-100, G-150 gel filtration and its specific activity was 528.0 unit/mg. The optimum pH and temperature for the CGTase activity were 8.0 and 80oC, respectively. The enzyme was stable in pH 8.0~11.0 at 60~80oC. The activity of purified enzyme was inhibited by Pb2+, Hg2+ and Zn2+. When CGTase was treated with each 20.5 unit, 41 unit, 205 unit and 410 unit to investigate the transglucosylation to stevioside by purified cyclodextrin glucanotransferase, transglucosylation rate to stevioside was 74.9%, 75.7%, 68.7% and 57.9%. Brown effect was observed above the concentration amounting to 205 unit of our CGTase.

St. John’s wort(Hypericum perforatum L.)의 생리활성 효과

조영제,천성숙,윤소정,김정환 한국응용생명화학회 2005 Journal of Applied Biological Chemistry (J. Appl. Vol.48 No.1

The physiological activity of St. John’s wort extracts were examined. Total phenol contents in the ethanol extracts (246.010.5 µg/ml) with St. John’s wort leaf was higher than that in water extract (237.413.2 µg/ml). The electron donating ability in the water extracts and in the ethanol extracts were 95.0% and 95.2% respectively. Antioxidant protection factor of the ethanol extract was higher than that of the water extract. The water extract from St. John’s wort leaves did not show an antimicrobial activity against Helicobacter pylori, but the ethanol extract revealed high antimicrobial activities such as 11 mm of clear zone in 100 µg/ml of phenol content and 13 mm of clear zone in 150 µg/ml of phenol content. The hot water extract showed an angiotensin converting enzyme inhibitory activity of 19.2%. The xanthin oxidase inhibitory activity of hot water and ethanol extract were very high, amounting to 84.8% and 100% respectively. The results suggested a possibility for developing the phenol compounds in St. John’s wort as anti Helicobacter pylori, anti-oxidant and anti-gout agents.