• 한국미생물·생명공학회지
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• 제27권6호
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• pp.471-476
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• 1999

Inulin fructotransferase(depolymerizing)(EC 2.4.1.93)(inulaseII) which converts inulin into di-D-fructofuranose-1,2':2,3'-dianhydride (DFAIII) was purified from Arthrobacter ureafaciens KCTC 3387 using column chromatography on DEAE-Toyopearl 650M and gel filtration of Sephadex G-200. The enzyme was purified 7-fold with a yield of 11% from a culture supernatant. The purified enzyme gave a single band on polyacrylamide gel electrophoresis, and the molecular weight of the enzyme was estimated to be 45,000 by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the enzyme reaction were pH6.5~7.0 and $55{\circ}C$, respectively. The enzyme was stable within a pH range of 5.0 to 10.6 and up to $60^{\circ}C$. The Km of this enzyme for DFAIII production was 11.9mM. The enzyme was inactivated by $Hg^{2+}$ and after exhaustive digestion of inulin by this enzyme, 1-kestose and nystose were produced in addition of DFAIII.

• 한국미생물·생명공학회지
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• 제11권1호
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• pp.47-52
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• 1983

Aspergillus sp.C-58 균주가 생산하는 extracellular inulase에 대하여 pH, charcoal처리 및 ammonium sulfate로 분별염석한 후 DEAE- cellulose를 이용한 column chromatography에 의하여 3개의 효소단백질(Peak I, II, III)로 분획되었으며 그 비율은 31. 1 : 1.7 : 1이였다. P- I, II의 I/S는 그 비율이 0.23 및 0.24로 거의 동일하였으나 P-III는 1.1로 P-I및 P-II와 상이하였다. Peak I 효소에 대하여 DEAE-Sephadex A-50을 이용한 ion exchange chromatography에 의하여 추출효소에 비교하여 약 408배 정제되었으며 다시 Sephadex G-75 및 Sephadex G-100에 2회 gel filtration하여 약 482배 정제되었다. 이상과 같이 정제한 Peak I의 효소액은 poly acrylamide를 이용한 disc gel electrophoresis 및 ultra centrifugation에 의하여 단일 단백질로 확인되었다.

• Applied Biological Chemistry
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• 제23권4호
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• pp.211-217
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• 1980

강력(强力)한 inulin 분해력(分解力)을 가진 Streptomyces chibaensis균주(菌株)를 이용하여, 효소생산(酵素生産)에 효과적인 배양조건 및 배지조성을 검토(檢討)하여 다음과 같은 결과를 얻었다. 1. 효소생산배지(酵素生産培地)의 최적 초기 pH는 7.5였으며, 이 조건하에서 $30^{\circ}C$에서 84시간 배양했을 때 가장 높은 효소역가를 나타냈다. 2. Inulase를 다량생산(多量生産)하기 위한 배지의 탄소원(炭素源)으로서는 다른 탄소원(炭素源)에 비하여 inulin이 가장 효과적이었다. 3. 무기질소원(無機質素源)으로는 $(NH_4)_2HPO_4$가 inulase생산(生産)에 가장 효과적(效果的)이었고, 유기질소원(有機窒素源)으로는 corn steep liquor가 효과적(效果的)이었다. 4. 무기염(無機鹽)으로 KCI 0.01%, $MgSO_4$ 0.05%, 그리고 $FeSO_4$ 0.0001% 첨가하였을 때 가장 효과가 컸다. 5. 탄소원(炭素源)(inulin) 1%와 질소원(窒素源)(corn steep liquor) 2%의 비율(比率)로 혼합(混合)하여 만든 배지(培地)에서 가장 효소생산(酵素生産)이 높았다.

• 한국미생물·생명공학회지
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• 제25권3호
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• pp.277-285
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• 1997

Kluyveromyces marxianus var. marxianus isolated as an inulin-assimilating microorganism produces inulin fructotransferase (inulaseII) which catalyses the conversion of inulin into di-D-fructofuranose 1, 2' : 2, 3' dianhydrde (DFAIII). The DFA produced by the organism was isolated by using active carbon column, and identified as DFAIII by high performance liguid chromatography. The culture medium giving maximum inulaseII production was found to consist of 1% sucrose and 0.75% yeast nitrogen base (YNB). The inulasell production was induced by inulin or sucrose as a carbon source and increased by addition of YNB as a nitrogen source. Optimal initial pH of the culture medium, culture temperature and medium volume for the enzyme production were pH 4.7, 30$\circ$C and 140 ml, respectively. Under the optimal conditions described above, the enzyme activity in the culture supematant reached 4.2 units/ml after cultivation for 36 h. The DFAIII was accumulated at 13.25 mg/ml after 48 h of culture in the Jerusalem artichoke tuber medium.