Biochemical Properties of Starch Granule Non - Digestive Enzyme ( SGNA ) of Bacillus polymyxa No.26

SOHN,CHEON -BAE,MYUNG - HEE KIM,JUNG - SURL,BAE,CHEORL - HO KIM 한국미생물생명공학회 1992 Journal of Microbiology and Biotechnology Vol.2 No.3

β - Amylase System Capable of Hydrolyzing Raw Starch Granules from Bacillus polymyxa No . 26 and Bacterial Identification

SOHN,CHEON -BAE,MYUNG - HEE KIM,JUNG - SURL,BAE,CHEORL - HO KIM 한국미생물생명공학회 1992 Journal of Microbiology and Biotechnology Vol.2 No.3

호알칼리성 Bacillus sp. No. 4의 Cyclodextrin Glycosyltransferase에 의한 Glycosyl Sucrose의 생산과 저충치성 당으로서의 응용

손천배,유미경,김명희,문숙경 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-

Action of a cyclodextrin glycosyltransferase (CGTase) produce from alkalophilic Bacillus sp. No. 4 was studied in a solution containing starch and sucrose to prepare glycosyl sucrose syrup with good sweetness and antidecaying properties of teeth. In the initial stage of the reaction the CGTase produced cyclodextrin, however, the cyclodextrin disappeared and glycosyl sucrose was formed with the lapse of reaction time. The best proportion of sucrose to starch for prodution of glycosyl sucrose was about 1 : 1. The optimum pH and temperature of the coupling reaction was pH 6.0 and 60℃, respectively. Main composition of glycosyl sucrose syrup prepared with 20% starch and 20% sucrose was sucrose 18%, glucosyl sucrose (G_2F) 15.3% and amltosyl sucorse (G_3F) 11.3%. And glucose, maltose and maltotriose were produced very little. Smaller amounts of acid and insoluble glucan were formed in the syrup by Streptococcus mtans OMZ176 than in the sucrose. Therefore, the prepared glycosyl sucrose sucrose syrup is expected to prevent teeth from decaying.

Pullulanase를 생산하는 Aeromonas caviae No. S-76의 특성과 배양조건

손천배,김명희,이명자 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-

A bacterial strain No. S-76 which produced pullulanase powerfully was isolated from soil. The isolated bacterium was 0.4∼0.6×0.8∼1.4 ㎛ in size, gram negative, rods, motile and was identified as Aeromonas caviae by Bergey's manual of determinative bacteriology with various characteristics investigated. The highest yield of pullulanase of the strain was obtained by using the following medium: 1% pullulan, soluble starch or corn starch as a carbon sources and 0.5% yeast extract, peptone as nitrogen sources with an initial pH of 9.0. The optimal culture conditions for production of pullulanase were at 32℃ for 2 days.

Aeromonas caviae No. S-76이 생산하는 Pullulanase의 정제, 특성 및 Maltosyl-β-Cyclodixtrin의 합성

손천배,김명희,이명자 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-

The crude enzyme solution obtained by shaking culture of Aeromonas caviae No. S-76 isolated from soil as pullulanase producing bacterium was purified by 50 folds with 21% yield by salting out with ammonium sulfate and column chromatography using DEAE-Sephadex A-50 and Sephadex G-150. The purified pullulanase had a molecular weight of 118,000 approximately by SDS-polyacrylamide slab gel electrophoresis and pI of 4.3 by isoelectric focusing. And optimum reaction temperature and pH for pullulanase were 50℃ and 8.0, respectively. The purified enzyme was relatively stable at pH 6.0∼9.0 and below 45℃. This enzyme synthesized maltosyl-β-cyclodextrin from mixture of β-cyclodextrin and maltose.

黑麴菌의 變異에 있어서 生澱粉糖化 酵素活性의 變動

孫天培 충남대학교 자연과학연구소 1987 忠南科學硏究誌 Vol.14 No.1

This study was to induce mutant irradiated with UV rays from Soju brewing black-Koji molds-Aspergillus awamori that produces highlv raw starch saccharyfying enzymes, and the obtained mutant was investigated for fungal characters, condition of enzyme production, characteristics of purified enzyme. The mutant strain showed morphological characteristics such as smaller conidiospore, vesicle, conidial head, and lighter conidia compared with the parent strain. Also when it was cultivated in wheat bran media, the enzyme productivity was beat 2-3 days at 30℃. The raw starch saccharifying enzyme activity was increased approxymately two times and acid protease activity was increased seven times. By the enzyme purification, two glucoamylases were separated. The peak Ⅰ showed gelatinized starch saccharifying activity but the peak Ⅱ showed both raw and gelatinized starch saccharifing activity. The molecular weight, and the optimum pH of the raw starch saccarifying enzyme were 94,000, 3.5, and it was stable at pH 2-9. The amount of sugars in the carbohydrate - protein link - age was 17% and they were identified as mannose, glucose, and galactosamine. When various kinds of raw starches were saccharified by using enzyme which had same gelatinized starch saccharifing activity, it was found that the mutant strain saccharified faster than the parent strain. The saccharifying speed of the parent enzyme was in the order of corn>wheat>cassava>sweet potato>potato starch, but in the mutant strain enzyme, cassava starch saccharified faster than wheat starch. The acid protease produced from the mutant strain did not modify with raw starch saccharifying enzyme and it did not affect enzyme activity.

Bacillus brevis CD162 Cyclodextrin Glycosyltransferase의 정제 및 특성

김명희,손천배,임영희,오태광 대전대학교 이과대학 기초과학연구소 1997 自然科學 Vol.- No.-

Bacillus brevis CD162가 생산하는 cyclodextrin glycosyltransferase (CGTase)를 ammonium sulfate 침전, DEAE-Sephadex CL-6B 및 Sephadex G-150 컬럼 크로마토그래피를 수행하여 분리정제하였다. 정제된 CGTase는 분자량이 약74.000, 등전점은 약 6.3인 단백질이었고, 정제된 단백질을 SDS-PAGE한 후 변성된 단백질을 재활성시켜 zymogram을 수행한 결과 cyclodextrin glycosyltransferase임을 확인할 수 있었다. 이 효소의 최적활성 pH와 온도는 각각 8.0과 55℃이었으며, pH 5.5~9.0과 50℃까지 안정한 활성을 보였다. 또한, CGTase의 NH₂-말단 부위의 아미노산서열은 Ser-Val-Thr-Asn-Tyr-Ser-Lys-Asp-Val-Ile-Tyr-Gln 이었으며, 전분으로부터 cyclodextrin으로의 전환률을 분석한 결과, α-cyclodextrin은 1.3%, β-cyclodextrin은 33.9%, γ-cyclodextrin은 9.7% 이었다(1997년 7월 10일 접수, 1997년 9월 25일 수리) The cyclodextrin glycosyltrasferase (CGTase, EC 3.2.1.19) from Bacillus brevis CD162 was purified by precipitating with ammonium sulfate, DEAE-Sepharose CL-6B column chromatography and Sephadex G-150 column chromatography. The molecular mass and pI of the purified enzyme were estimated to be 74,000 and 6.3 by SDS-PAGe and isoelectric focusing, respectively. The purified enzyme was clearly identified as the CGTase by zymogram after SDS-PAGE. The optimum pH and temperature for the enzyme activity were 8.0 and 55℃, respectively. The enzyme was stable at the range of pH 5.5~9.0, and up to 50℃. The amino acid sequence from the NH₂-terminal of the purified CGTase was Ser-Val-Thr-Asn-Lys-Val-Asn-Tyr-Ser-Lys-Asp-Val-Ile-Tyr-Gln. The yields of the products from starch as the substrate were 1.3% for α-,33.9% for β-, and 9.7% for γ-cyclodextrin.

Γ-Cyclodextrin Glycosyltransferase 생산균주의 분리, 동정 및 효소 생산조건

김명희,손천배,임영희,배경숙,오태광 대전대학교 이과대학 기초과학연구소 1997 自然科學 Vol.- No.-

Cyclodextrin glycosyltransferase 생산균주를 토양으로부터 분리하여 형태학적, 생화학적 그리고 균주의 세포벽 지방산 조성분석에 의해 Bacillus brevis로 동정하였고, Bacillus brevis CD162로 명명하였다. 또한 배지조성에 따른 cyclodextrin glycosyltransferase의 최적생산조건을 검토한 결과, 2.0% soluble starch, 0.75% yeast extract, 0.5% bacto peptone, 0.2% KHPO₄, 0.05% MgSO₄·7H₂O, 1.5% Na₂CO₃ (pH 10.2)의 배지 조건에서 30℃에서 96시간 배양하였을 때 가장 높은 효소생산인 0.9 unit/ml을 얻을 수 있었다.(1997년 7월 10일 접수, 1997년 11월 21일 수리) A cyclodextrin glycosyltransferase-producing bacterium was newly isolated from soil using alkaline pH medium containg 1% Na₂CO₃, The isolated strain was identified as Bacillus brevis by morphological and biochemical characteristics, and designated Bacillus brevis CD162. The strain showed the best enzyme production of 0.9 unit/ml after 96 hrs of culture at 30℃ in a medium of 2.0% soluble starch, 0.75% yeast extract, 0.5% bacto peptone, 0.2% K₂HPO₄, 0.005% MgSO₄·7H₂O and 1.5% Na₂Co₃at initial pH 10.2

무증자 흑미를 이용한 적색주의 제조

손천배 ( Cheon Bae Sohn ),박은배 ( Eun Bae Park ),문숙경 ( Suk Geang Mun ),김명희 ( Myeong Hee Kim ),장순애 ( Sun Ae Jang ),권종숙 ( Jong Suk Kwon ),손상혁 ( Sang Hyeok Sohn ) 충남대학교 생활과학연구소 2004 忠南生活科學硏究誌 Vol.17 No.-

Bacillus firmus Cyclodextrin Glycosyltransferase의 정제 및 특성

손천배(Cheon-Bae Sohn),김성애(Seong-Ai Kim),박영아(Young-A Park),김명희(Myung-Hee Kim),문숙경(Sook-Kyung Moon),장순애(Sun-Ae Jang),이명선(Myung-Sun Lee) 한국식품영양과학회 1997 한국식품영양과학회지 Vol.26 No.2

Bacillus firmus가 생산하는 cyclodextrin glycosyltran-sferase(CGTase)를 ammonium sulfate 침전, DEAE-Sephadex A-50 및 Sephdex G-100 column chromatography로 분리, 정제하였다. 이 방법으로 수율은 12%, 정제도는 49배인 SDS-PAGE 상에서 단일 band의 효소 단백질을 얻을 수 있었다. 정제된 CGTase는 Phast system의 SDS-polyacrylamide gel 전기영동에 의하여 분자량은 80,000dalton, isoelectric focusing으로 등전점은 9.6으로 추정되는 단백질이었다. 이 효소의 최적 활성 pH와 온도는 8.0,65℃이였으며, pH와 열안정성은 pH 5.5~9.0,50℃이였다. Soluble starch를 기질로 하여 24시간 효소반응한 액의 α- : β- : γ-cyclodextrin의 생성비율은 0.01 : 2.90 : 1.00으로 β- 및 γ-cyclodextrin을 주로 생산하였다. The cyclodextrin glycosyltransferase(EC 3.2.1.19) from Bacillus firmus was purified by precipitating with ammonium sulfate followed by, DEAE-Sephadex A-50 column chromatography and Sephadex G-100 column chromatography. In this way, we were able to obtain the single band protein on SDS-PAGE with a yield of 12%, whose purity was 49 fold. The purified CGTase was identified as a protein having molecular weight of approximately 80,000 dalton and isoelectric point of 9.6. The optimum pH and temperature for the enzyme activity were 8.0 and 65℃, respectively. The enzyme was stable at between pH 5.5 and 9.0 and up to 50℃. After 24hr of enzyme reaction using soluble starch as substrate, the ratio of α-, β- and γ-cyclodextrin production was 0.01 : 2.90 : 1.00, respectively. And this CGTase pro-duced mainly β- and γ-cyclodextrin.