• Journal of Ginseng Research
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• 제14권1호
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• pp.14-20
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• 1990

In An invertase (EC 3.2.1.26) was extracted from Korean giseng (Panax ginseng C.A. Meyer) with distilled tvater The ginseng invertase was purified about 62.6 folds purified by procedures including ammonium sulfate fractionation , DEAE-cellulofine chromatography and gelfiltrations through Sephadex G-75 and the recovery of enzyme activity was 11.1%. The homogeneity of the purified enzyme was probed by polyacrylamide gel disc electrophoresis. The purifled enzyme was divided into two different subunits by treating with a mixture of SDS and 2-mercautoethanol, and the molecular weight of the large subunit was estimatedtobe 116,000 and that of the small one to be 14,000. The optimal VH and temperature of the enzyme were pH 6 and 45$^{\circ}C$, respectively. The enzyme hydrolyzed specifically the hydrolyzation of the -fructofuranosides such as sucrose, raffinose and inulin. The Km values of the enzyme for sucrose and raffinose were determined to be 0.85 and 0.6 mM, respectively.

• 한국식생활문화학회지
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• 제5권2호
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• pp.269-274
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• 1990

건시제조 중 invertase의 활성 변화를 조사하고 정제된 invertase의 효소적 특성에 관하여 실험한 바 그 결과는 다음과 같다. 건조가 진행됨에 따라 invertase의 활성은 증가하여 건조 10일째 최고 활성을 나타내었으며 그 이후는 감소하였다. 건시에서 invertase를 250 mM potassium phosphate(pH 7.4)로 추출하여 $(NH_4)_2SO_4$염석, DEAE-cellulose 및 sephadex G-200 column chromatography의 과정으로 정제한 결과 정제도는 조효소액보다 약 27배 증가되었으며 회수율은 약 11%였다. 최적 pH는 sucrose, raffinose에 대하여 각각 5.0과 6.0으로 나타났으며 최적온도는 $40^{\circ}C$였다. 열과 pH 안정성은 $50^{\circ}C$까지 그리고 pH 5 부근에서 안정하였다. Sucrose에 대한 $K_m$값은 2.5 mM이었으며, 정제된 invertase는 polyacrylamide gel 전기영동상에서 하나의 band를 나타내었다.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.368-375
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• 1990

세포내 및 세포벽 뿐만 아니라, 세포외에도 invertase를 구성적으로 생산하는 효모 Rh.glutinis K-24로 부터 세포외 invertase를 disc 전기 영동상으로 단일한 상태로까지 정제하였다. 정제 효소의 효소화학적 성질을 밝힌 후 이미 보고한 바 있는 세포내 및 세포벽 invertase와 그 개락적 성질을 비교 검토하였다.

• Journal of Ginseng Research
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• 제14권1호
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• pp.21-26
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• 1990

The chemical composition and stabilities of the purified ginseng invertase were investigated. The purified enzyme was found to be a glycoprotein composed of 80.2% protein and 19.7% total sugar. The protein component of the enzyme was composed of acidic amino acid (9.3%), basic amino acid (48.9%), nonpolar amino acid (21.4%), polar amino acid (20.4%) and 6.1% S-containing amino acid. It showed especially high contents of histidine and serine. The enzyme was inactivated almost completely by the treatment with some proteases (papain, pepsin. trypsin, pancreatin and microbial alkaline pretense) and protein denatllrants (8M urea and 6M guanidine-HC1), bolt not with glyrosidase (${\alpha}$-amylase, ${\beta}$-amylase. glcoamylese and cellullase). btonosaccharides sllch as glilrose, fructose, galactose and mannose did not exert any influence on the enzyme activity. The activity of the enzyme was inhibited by Ag+, Mn2+, Hg2+, Zn2+ and Al3+, whereas Ca2+, Mg2+, Ba2+ and Fe3+ gave rather activating effects on the enzyme activity. The enzyme was relatively stable in the VH range of VH 6 and 8, and at the temperatures below 35$^{\circ}C$.

• Preventive Nutrition and Food Science
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• 제2권3호
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• pp.250-254
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• 1997

The internal invertase of Rhodosporidium toruloids mating type a cells was purified to a single band on SDS-PAGE from cell-free extract by acid precipitation, ion exchange chromatogaphy andgel filtration. The determined molecular weight of he purified enzyme was about 95,000 by gel filtration and 100,000 daltons on SDS-polyacryamide gel electrophoresis. This enzyme didn't show any activity change by several metal ions except 15.4% decrease by {TEX}$Mn^{2+}${/TEX} and was strongly inhibited by 2-mercaptoethanol and SDS. The invertase maintained its activity at high level until 70℃, but inactivated at 80℃ almost completely. The optimal temperature and pH of the enzyme were about 60℃ and pH 5.0, respectively. The stable pH range of invertase was narrow from pH 3.0 to 6.0. The Km value and isoelectric point of enzyme were 3.4×{TEX}$10^{3}${/TEX} M, pH 4.4, respectively.

• 한국식품영양학회지
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• 제2권2호
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• pp.8-13
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• 1989

A strain of Saccharomyces cerevisiae BY-366 was found to produce a strong sucrose-hydrolyzing enzyme Using this strain, the optimal culture conditions for the production of invertase were investigated. The results are as follows : 1. For enzyme production, optimal temperature, initial pH and critical concentrations of sucrose and raffinose were 3$0^{\circ}C$, 5.0 and 3.0%, respectively. 2. Enzyme production was reached maximum by organic nitrogen source, 0.3% yeast extract plus 0.5% bactopeptone. 3. It was appeared the presence of 0.1 M Mn2+ and Fe2+ ion was essential factors, on the other hand, 0.1 M Ag+ and Hg2+ ion almost block in yeast growth and enzyme production. 4. Invertase productivity was reached maximum within 3 days on stationary culture with medium-composed of sucrose 3%, bactopeptone 0.5%, yeast extract 0.3%, KEHPO. 0,1%, MgSO4.7H2O 0.05%.

• KSBB Journal
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• 제25권1호
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• pp.79-84
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• 2010

In the present study, we isolated a new bacterial strain producing invertase (EC 3.2.1.26) and determined optimized culture condition in flask culture. The strain was identified as Bacilus flexus determined by the 16S rDNA sequencing method. The invertase was produced only in the sucrose medium as the sole carbon source. Potassium nitrate was an adequate nitrogen source for enzyme production, whereas meat peptone showed the highest bacterial growth. Enzyme production was increased about 2-fold when $MgSO_4\cdot7H_2O$ was supplemented to the growth media. The optimum temperature was found to be $30^{\circ}C$ for both enzyme production and bacterial growth. Invertase exhibited pH optima in the range 5.0-6.0 and have a temperature optimum at $40^{\circ}C$, similarly to other invertases found from different microbial sources. Several mineral ions (K and Fe) stimulated the invertase activity, whereas some bioelements (Ag, Mg, and Mn) inhibited enzyme activity. Under the optimized culture condition, the maximum enzyme production (over 250 units/mL) was achieved at 20 h. To the best of our knowledge, this is the first time to report on invertase production by Bacilus flexus.

• 한국환경과학회지
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• 제4권4호
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• pp.21-21
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• 1995

여러 파장의 광선(백색, 적색, 녹색 그리고 청색광)과 NAA, $GA_3$, BA 등의 식물호르몬을 옥수수와 녹두 유식물의 잎에 처리하여 환원당 축적과 invertase isozymes 활성에 미치는 효과를 조사하였다. NAA는 옥수수와 녹두 잎의 환원당과 invertase isozymes 활성의 증가를 촉진하였으나, $GA_3$는 환원당 축적과 효소의 활성증가에 효과적이지 못하였다. 한편 BA는 녹두 잎의 imvertase isozmes의 활성증가를 유도하는 반면, 옥수수 잎에 있어서는 큰 영향을 미치니 못하였다. 환원당의 축적에 있어서 적색광은 백색광에 비하여 효과적이였으나, invertase isozymes의 활성증가에 있어서는 여러 파장의 광선 모두가 촉진효과를 보여주지 않았다. 식물호르몬과 광선의 동시처리에 있어서 NAA와 백색광 동시처리는 녹두 잎의 환원당 축척 및 invertase isozymes의 활성증가에 매우 효과적이였으며 NAA와 청색광 동시처리는 옥수수 잎의 환원당 축척과 invertase isozymes의 활성증가에 매우 효과적이었다. 이상의 결과로 보아 옥수수와 녹두 잎의 환원다 축척 및 inverses isozymes의 활성증가에 있엇 식물호르몬, 큭히 NAA는 여러 파장의 광선보다 더 중요한 요인인 것으로 사료된다.

• 한국환경과학회지
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• 제4권4호
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• pp.323-333
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• 1995

여러 파장의 광선(백색, 적색, 녹색 그리고 청색광)과 NAA, $GA_3$, BA 등의 식물호르몬을 옥수수와 녹두 유식물의 잎에 처리하여 환원당 축적과 invertase isozymes 활성에 미치는 효과를 조사하였다. NAA는 옥수수와 녹두 잎의 환원당과 invertase isozymes 활성의 증가를 촉진하였으나, $GA_3$는 환원당 축적과 효소의 활성증가에 효과적이지 못하였다. 한편 BA는 녹두 잎의 imvertase isozmes의 활성증가를 유도하는 반면, 옥수수 잎에 있어서는 큰 영향을 미치니 못하였다. 환원당의 축적에 있어서 적색광은 백색광에 비하여 효과적이였으나, invertase isozymes의 활성증가에 있어서는 여러 파장의 광선 모두가 촉진효과를 보여주지 않았다. 식물호르몬과 광선의 동시처리에 있어서 NAA와 백색광 동시처리는 녹두 잎의 환원당 축척 및 invertase isozymes의 활성증가에 매우 효과적이였으며 NAA와 청색광 동시처리는 옥수수 잎의 환원당 축척과 invertase isozymes의 활성증가에 매우 효과적이었다. 이상의 결과로 보아 옥수수와 녹두 잎의 환원다 축척 및 inverses isozymes의 활성증가에 있엇 식물호르몬, 큭히 NAA는 여러 파장의 광선보다 더 중요한 요인인 것으로 사료된다.

• Journal of Plant Biology
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• 제38권3호
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• pp.251-258
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• 1995

The soluble acid invertase ($\beta$-D-fructofuranoside fructohydrolase, EC 3.2.1.26) was isolated and characterized from the hypocotyls of mung bean (Phaseolus radiatus L.). The enzyme was purified to apparent homogeneity by consecutive step using diethylaminoethyl (DEAE)-cellulose anion exchange, Concanavalin (Con) A affinity and Sephacryl S-300 chromatography. The overall purification was about 148-fold with a yield of about 15%. The finally purified enzyme exhibited a specific activity of about 139 $\mu$mol of glucose produced mg-1 protein min-1 at pH 5.0 and appeared to be a single protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing PAGE. The enzyme had the native molecular weight of 70 kD and subunit molecular weight of 70 kD as estimated by Sephadex G-200 chromatography and SDS-PAGE, respectively, suggesting that the enzyme was composed of a monomeric protein. On the other hand, the enzyme appeared to be a glycoprotein containing N-linked high mannose oligosaccharide chain on the basis of its ability to bind to the immobilized C on A. The enzyme had a Km for sucrose of 1.8 mM at pH 5.0 and maximum activity around pH 5.0. The enzyme showed highest enzyme activity with sucrose as substrate, but the activity was slightly measured with raffinose and cellobise. No activity was measured with maltose and lactose. These results indicate the soluble acid invertase is a $\beta$-fructofuranosidase.