• 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.

• Korean Journal of Food Science and Technology
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• v.11 no.4
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• pp.283-290
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• 1979

Partially purified ${\beta}-fructosidase$ (inulase) from Kluyveromyces fragilis was immobilized on Tygon tube and aminoethyl-cellulose, respectively and both preparations were characterized. Silanization of Tygon tube in chloroform at $65^{\circ}C$ and treatment with 10 % glutaraldehyde were critical for the immobilization of inulase on Tygon tube, while 2 % glutaraldehyde was effective for the immobilization on aminoethyl-cellulose. The derivative of Tygon tube showed 11.5 units of inulase activity per g of dried matrix with retention of 22.5 % of original activity against inulin, whereas one of aminoethyl-cellulose showed 39.3 units per g of dired matrix with 53.4 % of retention. Studies of enzyme stability, pH and temperature dependences, and $K_m$ values are presented for inulase and invertase activities of both immobilized enzymes.

• Applied Biological Chemistry
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• v.23 no.4
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• pp.211-217
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• 1980

Using Streptomyces chibaensis, which produces a strong inulin-hydrolyzing enzyme, the optimum cultural conditions and composition of the medium for the production of inulase were studied. 1. The highest enzyme activity was obtained at pH 7.5 after 84 hours culture at $30^{\circ}C$. 2. None of carbon source better than inulin was found. 3. $(NH_4)_2HPO_4$ and corn steep liquor were favourable inorganic and organic nitrogen sources for the production of inulase. 4. $KCl,\;MgSO_4\;and\;FeSO_4$ as the metallic salts were effective for the enzyme production at their concentrations of 0.01, 0.05 and 0.0001%, respectively. 5. The highest production of inulase was obtained from the medium of inulin 1.0% and corn steep liquor 2.0% concentrations, respectively.

• Applied Biological Chemistry
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• v.27 no.1
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• pp.45-51
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• 1984

Kluyveromyces marxinus LG contains an inulase activity which is inducible by growth on inulin. The highest enzyme activity obtained at the initial stage of stationary phase of growth curve. The partially purified inulase hydrolyzed inulin, sucrose and raffinose. This enzyme showed the maximal activity at pH 4.0 and temperature of $55^{\circ}C$ for inulin and sucrose. It was also markedly inhibited by $Hg^{++}\;and\;Ag^{+}$ but not EDTA. This inulase was charaterized as a undo type which contains invertase activity and Km values for inulin and sucrose were $1.2{\times}10^{-2}M\;and\;1.3{\times}10^{-4}M$, respectively.

• Korean Journal of Food Science and Technology
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• v.25 no.6
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• pp.780-786
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• 1993

About 7.7% of the total anaerobic bacteria in pig feces grew with clear zone around the colonies on the agar medium containing inulin as a sole carbon source. Among these bacteria, a strain with the strongest inulin-utilizing activity was isolated and identified as Bacteroides sp. based on its morphological and taxonomical characteristics. The isolate grew well with inulin, fructooligosaccharides or glucose as a sole carbon source, while its growth dropped to 50% of that obtained with glucose when soluble starch or sucrose are used. Since the inulase activity was found only when fructooligosaccharides or inulin was added to the growth medium, but not when glucose, sucrose or soluble starch was applied, the inulase production was considered to be induced by fructooligosaccharides or inulin. The highest inulase activity, 0.42 U/ml was detected with the inulin medium and 0.25 U/ml with fructooligosaccharides medium. The cell growth of the isolated strain increased with the amounts of inulin up to 2%(w/v) and maximum production of inulase was found in the cells fed 1% inulin. The inulase of the isolated Bacteroides sp. showed its maximum activity at pH $7.0{\sim}7.5\;and\;50{\sim}50^{\circ}C$ and was found to be an exoinulase judging by its mode of action.

• Applied Biological Chemistry
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• v.18 no.1
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• pp.42-51
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• 1975

Penicillium sp I which produces a powerful hydrolysing enzyme was isolated from putrefid and dry Jerusalem artichoke medium. The strain was used to study on the optimum culture conditions for enzyme production. The results obtained are as follows: 1. Penicillium sp I was a vigorous strain to produce inulase. 2. The optimum culture conditions of the strain was examined in the Jerusalem artichoke extract medium and the synthetic medium. 3. Inulase productivity in the Jerusalem artichoke extract medium was higher than that of the synthetic medium. 4. The optimum culture period of the Jerusalem artichoke extract medium was four days, whereas that of the synthetic medium was five days. 5. The optimum temperature, pH and concentration in the Jerusalem artichoke extract medium were $30^{\circ}C$, 5.0 and 4.0% (W/V), respectively. Meanwhile, the optimum temperature, pH and concentration in the synthetic medium were $30{\sim}33^{\circ}C$, $5.0{\sim}6.0$, and $1.0{\sim}1.5%$ (W/V), respectively. 6. Corn steep liquor, peptone, $(NH_4)_2HPO_4,\;NH_4H_2PO_4,\;(NH_4)_2SO_4$, etc. were favorable as nitrogen sources. Of these, especially, Corn steep liquor and peptone as organic nitrogen sources caused an increase in inulase production in the synthetic medium. 7. All sugars except for inulin have no effect upon the inulase production. 8. KCl, $MgSO_4\;and\;FeSO_4$ were favourable mineral sources for inulase production.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.593-599
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• 1989

A strain producing extracellular endo-type inulase was selected from Actinomycetes isolated from soil, and identified as Streptomyces sp. The maximum inulase production was obtained with medium containing inulin 1.0%, yeast extract 1.0%, (NH$_4$)$_2$HPO$_4$ 0.4%, NH$_4$H$_2$PO$_4$0.8%, KCl 0.05%, MgSO$_4$ㆍ7$H_2O$ 0.05%, FeSO$_4$ㆍ7$H_2O$ 0.001% at 96 hours culture in jar fermentor. The endo-type inulase was considered to be an inducible enzyme produced by inulin only.

• Microbiology and Biotechnology Letters
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• v.1 no.1
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• pp.37-42
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• 1973

C-74 belonging to the genus of Aspergillus species was selected from two hundred and seventy microorganisms stored at the lab, of microbiology. 1. Sample Inulase was precipitated mainly at pH 8 to 5 by isoelectric point fraction method. 2. The optimum pH of Inulase activity of the enzyme from C-74 strain was about 3.0 3. The optimum temperature of Inulase activity of the enzyme from C-74 strain was about $55^{\circ}C$ 4. The Inulase from C-74 strain was stable at $50^{\circ}C$ for 10mins. 5. The range of the pH stability of the Inulase from C-74 strain was about 2.5-4.5 6. Effect of metal ions on the Inulase activity of the enzyme from C-74 strain was activated by Mg$\^$++/, Sb$\^$+++/ and inhibited by Ag$\^$++/, Hg$\^$++/.

• 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 $4.54{\times}10^{-4}M$.