• Applied Biological Chemistry
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• v.30 no.2
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• pp.169-178
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• 1987

The extracellular and intracellular inulases from Kluyveromyces marxianus were purified and characterized. The maximum production of both inulases was achieved at stationary phase in a pH-controlled medium at pH 5 with yeast nitrogen base as organic nitrogen source. Each enzyme was concentrated by tannic acid precipitation and separated into two fractions by DEAF-cellulose chromatography. Electrophoretic analysis showed that the four fractions had three glycoprotein bards each. Only main glycoprotein band, however, had both inulase and invertase activities. There were no significant differences between two enzymes in the optimum pH and temperature. But the intracellular inulases had higher heat stability and less affinity toward inulin than the extracellular enzymes do. All the purified enzymes were considered to be exo-inulases using hydrolyzate analysis with TLC.

• Microbiology and Biotechnology Letters
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• v.27 no.6
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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.

• Korean Journal of Food Science and Technology
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• v.13 no.1
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• pp.67-73
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• 1981

An inulase from Streptomyces chibaensis was purified about 120-fold with the yield of 38 by ethanol precipitation, DEAE-cellulose chromatography and Sephadex G-200 gel-filtration. The purified enzyme showed the maximal activity at pH 6.5 and was fairly stable between pH $5.0{\sim}9.0$ The enzyme was slightly activated by $Mn^{++},\;Mg^{++}\;and\;Co^{++}$, and markedly inactivated by $Hg^{++}\;and\;Ag^{+}$. The inulase was characterized as a typical endo-inulase which hydrolyzed inulin in a random manner and Km for the inulin was $4.54{\times}10^{-4}M$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.17 no.3
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• pp.205-210
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• 1988

The inulase(EC 3.2.1.7) was isolated from the tuber of Jerusalem artichoke by conventional purification methods including ammonium sulfate fractionation, Sephadex G-100 filtration, and DEAE-cellulose column chromatography. The enzyme was purified 6,470 fold with 42% recovery, The enzyme was consisted of a polypeptide of Mw 57,000. The optimum temperature and the optimum pH for the enzyme action was $33^{\circ}C$ and pH 5.0, respectively. The enzyme was highly specific for inulin as a substrate. The km for inulin was 20mM. The inulase was not a metalloenzyme and was inhibited completely by 10mM $Mg^{2+},Ca^{2+},\;or\;Hg^{2+}$.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.47-52
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• 1983

The extracellular inulase produced by Aspergillus sp. C-58 was isolated by pH and charcoal treatment, precipitation with ammonium sulfate from the crude extract, and separated into 3 fractions (P-I, II, III) by DEAE-cellulose column chromatography in the ratio of 31.1:1.7:1 with respect to the activity. The ratio of inulase activity to sucrase activity of P-I, P-II and P-III fraction was 0.23, 0.24 and 1.1 respectively. The enzyme P-I fraction was purified 482 fold with a 22.8% yield by DEAE-Sephadex A-50, Sephadex G-75, Sephadex G-100 (1st and 2nd) column chromatography, and appeared homogeneous on polyacrylamide disc gel electrophoresis and ultracentrifugation.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.484-489
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• 1990

An intracellular inulase ( fJ-fructofuranoside fructohydrolase, EC 3.2.1.26) from Bacillus subtilis has been partially purified and its mode of action and general properties were studied. The enzyme had an apparent molecular weight of 49,000 as estimated by gel filtration and its pI point was 5.2. Substrate concentration studies showed an apparent Km of 10 mM for sucrose and of 18 mM for raffinose. The enzyme was an acid-labile protein with a pH optimum of 6.6. The optimum temperature was 50$^{\circ}$C. The enzyme acts on straight chain oligo- and poly-fructosides of the inulin series via a exo-wise cleavage mechanism, as well as on sucrose.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.551-558
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• 1992

The extracellular endoinulase from Streptomyces sp. 556 was purified and characterized, The culture broth was fractionated by ammonium sulfate saturation followed by DEAE-cellulose column chromatography and 5ephadex G-200 gel filtration, The ultimately purified fraction revealed a single band in 7.5% polyacrylamide gel electropherogram. The purified enzyme showed the maximal activity at pH 5.5-6.0 and $50^{\circ}C$, but lost 93% of inulase activity after 30 min incubation at $55^{\circ}C$ . The essen.tial amino acid residue for catalytic activity appeared to be tryptophan. This endo inulase was activated by $Mn^{2+}$, whereas inactivated by $Ag^{+}$, $Hg^{+}$, $Cu^{2+}$, $Zn^{2+}$, $Fe^{3+}$ and $Mo^{6+}$ EDTA and 8-hydroxyquinoline inhibited the enzyme so that the enzyme was considered to be a metalloenzyme. The Km value for inulin was 0.287 mM, and no invertase or $\alpha$-glucosidase activity was found in the enzyme.