• KSBB Journal
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• 제8권4호
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• pp.313-319
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• 1993

다공성 실리카표변에 폴리에틸렌이민과 글루타르 알데히드로 전화당 효소를 고정화시키고 자당과 에 탄올로부터 메틸 프룩토시드를 합성한 결과 다음과 같은 결론을 얻었다. 다공성 설리카를 폴리에틸렌이 민으로 표면처리하여 얻어진 고정화 담체는 그 다공 성 구조와 표면화학적 특성이 효소의 고정화에 적합 하여 전화당 효소에 대한 고정화 함량 120mg/g, 활 성도 lOOD/mg을 얻어 배당체의 합성반응에서 필요 로 하는 고정화 촉매를 제조할 수 었었으며 고정화 담체 표변에 형성된 폴리에틸렌이민 피막은 효소 부근에 친수성 분위기를 유지하여 높은 효소활성을 나 타내었고 에틸 프룩토시드가 효소에 접근하여 가수 분해되는 반응을 억제하여 메틸 프룩토시드의 수율 과 농도를 크게 증가시켰다. 자당과 메탄올 수용액 으로부터 메틸 프룩토시드를 합성한 결과, 자당 농도 O.291mol/l , 메탄올 농도 30%(v/v), 반응온도 $25^{\circ}C$, pH 4.8, 효소활성도 2U/ml일 때 자당의 전화 율 91.2%, 에틸 프룩토사드 농도 27.0g/l , 그 평균 수율 55.9%을 얻었으며, 위의 연구결과로부터 고정화 효소에 의한 배당체의 합성반응이 연속생산공정 으로 연구개발 될 수 있을 것으로 기대된다.

• 한국식품영양학회지
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• 제2권2호
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• pp.14-20
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• 1989

A strain of Saccharomyces cerevisiae BY-366 was isolated to produce a strong sucrose-hydrolyzing enzyme. After entrapment of yeast cell invertase with alginate, enzymatic properties of immobilized cells were investigated. The results are as follows. 1. The optimum pH of invertase in immobilized cells and non- immobilized cells was 6.0 and 5.0, and pH stability of invertase in immobilized cells and non- immobilized cells was 6.0 and 5.0, respectively. 2 Activation energy of immobilized cells was 4.7 kcal/mol. 3 The immobilized preparation exhibited high resistance to heat and urea Induced denaturation. 4, The bead size less than 2 mm in diameter was desirable. 5. In spite of repeated use, the enzyme activity of immobilized cells was inhibited slightly in batch reaction, and a small column of the immobilized preparation could hydrolyze relatively high concentration of sucrose almost quantitatively to more than 6 days.

• 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.

• 한국미생물·생명공학회지
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• 제34권4호
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• pp.318-322
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• 2006

알칼리와 고온에 대한 내성을 가진 invertase를 대량생산하기 위하여 호알칼리성이며 고온성 세균인 Bacillus sp. TA-11의 세포내 invertase유전자를 pUC 19벡터를 이용하여 E. coli HB101에 클로닝 시키고 발현시켜 invertase를 강력하게 생산하는 재조합 E. coli (pYC17)를 얻었다. 재조합 E. coli(pYC17)를 이용한 invertase 생산 최적조건을 검토한 결과 E. coil(pYC17)를 0.25% sucrose, 0.5% yeast extract, 0.1% $K_2HPO_4$와 0.1% $KH_2PO_4$을 함유한 SY배지(초기 pH 9.0)에 접종하여 37$^{\circ}C$에서 9시간 배양하였을 때 친주보다도 많은 47.7 U/ml-cell free extract의 invertase가 생산되었다.

• 한국식품과학회지
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• 제12권3호
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• pp.176-181
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• 1980

Invertase의 고정화에 대해서 두가지 carrier matrix를 사용하여 연구하였다. Sepharose에 ${\omega}-aminohexylarm$를 붙인 후 효소를 결합시키는 indirect coupling method와 cellulose에 phenexyacetyl group의 linkage를 만들어 변형시킨 후 (modified cellulose), 여기에 효소를 흡착시키는 hydrophobic adsorption method로서 제조하였다. 각각의 고정화 수율(immobilized yield)은 ${\omega}-aminohexyl\;sepharose$의 경우 첨가한 효소의 26.0%의 activity를 고정화 시킬 수 있었으며, phenoxyacetyl cellulose의 경우는 72.9%였다. 제조한 고정화 효소의 안정성, ph영향, 온도 영향 및 Km값을 조사하였다. ${\omega}-aminohexyl\;Sepharose$에 고정화시킨 invertase근 최적 pH 4.5, 최적 온도 $60^{\circ}C$, 활성화 에너지 $5,941\;cal/mole{\cdot}deg$, Km값 22.2 mM이었으며 phenoxyacetyl cellulose에 고정화시킨 invertase는 최적 pH 4.0, 최적 온도 $60^{\circ}C$, 활성화 에너지 $7,769\;cal/mole{\cdot}deg$, Km 값 69.9mM이었다.

• Journal of Plant Biology
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• 제38권4호
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• pp.349-357
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• 1995

The alkaline 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 by consecutive step using diethylaminoethyl (DEAE)-cellulose anion exchange, 1st Sephadex G-200, DEAE-Sephadex A50 and 2nd Sephadex G-200 chromatography. The overall purification was about 77-fold with a yield of about 6%. The finally purified enzyme exhibited a specific activity of about 48 $\mu$mol of glucose produced mg-1 protein min-1 at pH 7.0 and appeared to be a single protein by nondenaturing polyacrylamide gel electrophoresis (PAGE). The enzyme had the native molecular weight of 450 kD and subunits molecular weight of 63 kD and 38 kD as estimated by Sephadex G-200 chromatography and SDS-PAGE, respectively, suggesting that the enzyme is a heteromultimeric protein composed of two types of subunits. On the other hand, the enzyme appeared to be not a glycoprotein according to the results of Con A chromatography and glycoprotein staining. The enzyme had a Km for sucrose of 19.7 mM at pH 7.0 and maximum activity around pH 7.5. The enzyme was most active with sucrose as substrate, compared to raffinose, cellobiose, maltose and lactose. These results indicate the alkaline invertase is a $\beta$-fructofuranosidase.

• 한국식품영양과학회지
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• 제19권6호
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• pp.577-582
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• 1990

Aspergillus niger로 부터 invertase 생성의 최적조건과 생성된 효소의 특성을 조사하였다. 곰팡이는 탄소원으로 inulin과 sucrose를, 질소원으로 yeast extract를 사용했을 때 최대효소 생성을 얻을 수 있었다. 또한 사용한 배지의 pH를 4.5, 배양온도를 3$0^{\circ}C$로 유지했을 때 효소생성이 높았다. 효소 활성을 위한 최적조건은 pH 5.0 및 5$0^{\circ}C$였다. 효소는 pH 6.0에서 가장 안정했으며, 또한 5$0^{\circ}C$이하에서 안정하였다. 효소는 Hg++, Ag+ 및 Cu++ 이온의 첨가에 의하여 불활성화 되었다.

• 한국식품영양과학회지
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• 제19권5호
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• pp.429-433
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• 1990

Production and properties of invertase from. Aspergillus niger were investigated. Inuli and sucrose were best carbon source and yeast extract was most suitable for the production of the enzyme among tested carbon and nitrogen sources. The enzyme was maximally produced by cultivating the organism at medium of pH 4.5 and temperature of 3$0^{\circ}C$ The optimum pH and temperature for the enzyme activity were pH 5.0 and temperature of 5$0^{\circ}C$ respectively Among tested metal ions Hg2+ Cu2+ and Ag+ ions inhibited the enzyme activity drastically.

• KSBB Journal
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• 제5권2호
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• pp.107-112
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• 1990

고정화단체로 제조된 다공성실리카를 3-aminopropyl-triethoxysilaneacetone 용액으로 표면처리하고 glutaraldehyde로 전화당효소를 공유결합시켜 진화당효소의 고정 화함량과 고정화활성도를 평가한 겨로가, 고정화함량 1200mg/ g, 활성도 26.9~ 70.2%로 우수한 고정화특성은 나타내였으며, 다상성살리카가 고정화담체로 활용되기 위한 구조직 특성을 가진 것으로 평가된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권3호
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• pp.167-172
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• 2003

Foam fractionation can be used to enrich a hydrophobic protein such as bromelain from an aerated dilute protein solution because the protein foams. On the other hand, a protein such as invertase, which is hydrophilic, is not likely to foam under similar aerated conditions. While a foam fractionation process may not be appropriate for recovering a hydrophilic protein alone, it is of interest to see how that non-foaming protein affects the foaming protein when the two are together in a mixture. The bromelain enrichment, activity and mass recovery were observed as a function of the solution pH in order to explore how invertase can affect the recovery of bromelain in a foam fractionation process.