• Proceedings of the Korean Society of Life Science Conference
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• 2002.03a
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• pp.90-90
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• 2002

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.259-264
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• 1988

Two forms of extracellular inulinase, designated as PI and PII were detected in the crude enzyme preparation from n species of Pseudomonas isolated from soil. PI and PII were purified to homogeneity by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, Sephadex G-100 and Sephadex G-200 gel filteration. Both isoenzymes catalyzed specifically and endowise the cleavage of the $\beta$-2,1-fructofranoside linkage of inulin, and displayed no action upon sucrose, raffinose and levan. The optimal pH values for the PI and PII enzyme were pH 5.5 and 6.0, respectively and the highest activity of the two enzymes was observed at 55$^{\circ}C$. The Km values of PI and PII were calculated to be 2$\times$10$^{-3}$M and 5$\times$10$^{-3}$M, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.4
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• pp.306-308
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• 2001

A recombinant human growth hormone(hGH) was expressed as a secretory product in the yeast Saccharomyuces cerevisiae. There different leader sequences derived from the mating fac-tor $\alpha$1(MF$\alpha$1) inulinase and invertase were used to direct the secretion of hGH into the extracel-lular medium. Among three leader sequences tested, the inulinase leader sequence was found to be the most efficient in the secretory expression of hGH. In contrast, no hGH was detected in the ex-tracellular medium with the invertase leader sequence. After 48 h shake-flask culture, the yields of hGH secreted into th emedium by the invertase. MF$\alpha$1 inulinase and invertase leader sequences were approximately 0, 0.3 and 0.9 mg/L, respectively. The secretion efficiencies were also found to be 0, 3.8 and 13% for the invertase , MG$\alpha$1 and inulinase leader sequences, respectively.

• Microbiology and Biotechnology Letters
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• v.20 no.1
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• pp.20-24
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• 1992

In order to reuse inulinase effectively, a method for immobilizing both endo- and exoinulinase to vinylsulfone activated agarose via covalent bond was investigated. The immobilized enzyme preparation had, respectively, 400 U for exoinulinase activity and 80 U for endoinu- Iinase activity per gram gel. A thermal stability by immobilization had increased in the case of exoinulinase. Optimum pHs for two immobilized enzymes were 4.4 to 5.0. Synergistic effect which depends on mixed ratio of two immobilized enzymes was the best when the mixed ratio of endo/exo lay between 0.1 and 0.5, and its activity of the mixed enzyme increased 1.7 times as compared to that of each immobilized enzyme. Inulinase activities of both of the immobilized enzymes did not change during 20 times experimental runs in a batch reactor.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.90-93
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• 1997

Recombinant exoinulinase was partially purified form the culture supernatant of S.cerevisiae by(NH4)2SO4 precipitation and PEG treatment. The purfied inulinase was immobilized onto Amino-cellulofine with glutaraldeyde as a cross-linking agent. Immobilization yield based on the enzyme activity was about 15%. Optimal pH and temperature of immobilized enzyme were found to be 5.0 and 6$0^{\circ}C$, respectively. The enzyme activity was stably maintained in the pH ranges of 4.5 to 6.0 at 6$0^{\circ}C$. 100% of enzyme activity was observed even after incubation for 24 hr at 6$0^{\circ}C$. In the operation of a packed-bed reactor containing 412U inulinase, dahalia inulin of 7.5%(w/w) concentration was completely hydrolyzed at flow rate of 2.0mL/min at 6$0^{\circ}C$, resulting in a volumetric productivity of 693 g-reducing sugars/L/h. Under the reaction conditions of 1.0mL/min flow rate with 2.5% inulin at 6$0^{\circ}C$, the reactor was successfully operated over 30 days without loss ofinulinase activity.

• Applied Biological Chemistry
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• v.39 no.2
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• pp.99-103
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• 1996

A bacterial strain producing a novel endo-inulinase, hydrolysing inulin into oligosaccharides was isolated from soil and identified as Arthrobacter sp. S37 The enzyme production was induced by inulin and jerusalem artichoke extract. The maximum enzyme production was obtained with medium containing 1.5% jerusalem artichoke extract, 1.0% yeast extract, $0.5%\;NaNO_3,\;0.05%\;MgSO_4{\cdot}7H_2O,\;0.05%\;KCl,\;0.0016%\;FeCl_3{\cdot}6H_2O\;and\;0.05%\;KH_2PO_4$. The optimum temperature and pH for the enzyme production were $30^{\circ}C$ and 8.0, respectively. Under the optimum condition, the enzyme activity in the culture broth reached at maximum, 10.8 units/ml after cultivation for 24 hours.

• Applied Biological Chemistry
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• v.40 no.1
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• pp.34-38
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• 1997

For the effective production of functional oligosaccharides(DP 3-5) from inulin in chicory extracts, the acid hydrolysis and enzymatic endo-inulinase reaction were compared. Acid hydrolysis was unfavorable ; the content of oligosacharides in total sugar increased to 26.0% for 12 min at $55^{\circ}C$ and 24.6% at 6 min at $65^{\circ}C$ and showed little change for 30 min. The content of high DP(DP 6) decreased from 83.5 to 49.5% and 23.0% for 30 min, repectively. Glucose, fructose and sucrose increased to 24.6% and 50.3%, respectively. Hydrolysis of chicory extracts with purified endo-inulinase from Arthrobacter sp. S37 was carried out at $40^{\circ}C$ and pH 7.5 for 44 hrs. The content of high DP($DP{\geq}6$) in total sugar decreased from 83.5 to 23.0% and that of inulobiose(F2) and DP 3-5 increased to 66.1%. Glucose, fructose and sucrose were not produced. The hydrolysis of chicory extracts without DP 1 and DP 2 with crude or with purified enzyme were also carried out. In contrast to the hydrolysate of crude enzyme, that of purified endo-inulinase did not contain glucose, fructose, sucrose, F2 and 1-kestose(GF2). The content of oligosaccharides in the hydrolysate of the purified endo-inulinase were 79.2%, composed mainly of inulotriose(F3), inulotetraose(F4) and inulopentaose(F5), which shows that the enzymatic hydrolysis using purified endo-inulinase from Arthrobacter sp. S37 is the best method for oligosaccharides production from inulin in chicory extracts.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1101-1106
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• 2010

Pulsed electric field (PEF) of 1Hz, 1.5 kV, and 1ms increased the activities of catalase and inulinase over the whole range of applied Se concentrations compared with the non-treated cultures. A significant effect of selenium concentration (in the range of 5-14 ${\mu}g/ml$) on both intra- and extracellular enzyme activities was noted. At a Se concentration of 10 ${\mu}g/ml$, the activities of intra- and extracellular inulinases and extracellular catalase in the PEF-treated cultures reached the maximum of 71 U/g d.m., 46 U/g d.m., and approx. 8 U/ml, respectively. The maximum activity of intracellular catalase of approx. 6 U/ml (with and without PEF) was recorded at 5 ${\mu}g$ Se/ml. Further increasing of selenium concentration caused a decrease in the activity of the enzymes.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.121-126
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• 1997

A bacterial strain LC-413, producing an extracellular inulin fructotransferase (depolymerizing) which converts inulin into di-D-fructofuranose dianhydride (DFAIII), was isolated from soil. Inulin fructotransferase from the isolate identified as a strain Flabobacterium sp. was purified from the culture broth by ammonium sulfate precipitation, followed by column chromatograpies on DEAE-Toyopearl 650 M and phenyl-Toyopearl 650 M. The purified enzyme gave a single band on an electrophoretic disc-gel. The molecular weight of the enzyme was estimated to be 44, 000 Da by SDS-polyacrylamide gel electrophoresis, and 45, 000 Da by gel filtration, suggesting the monomeric state of the enzyme. The isoelectric point of the enzyme was about pH 4.5. The optimal pH and temperature for the enzyme reaction were 6.0 and $50^{\circ}C$, respectively. The purified enzyme digested inulin into di-D-fructofuranose-l, 2': 2, 3'-dianhydride, confirming the enzyme was an inulin fructotransferase (inulinase II).

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.582-585
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• 2003

Sequence analysis indicated that Bacillus polymyxa MGL21 CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase. Furthemore, CFTase possessed a highly conserved core region. In order to understand the role of the core region on the function of CFTase from B. polymyxa MGL21 CFTase ${\Delta}NC$ was prepared. The molecular weight of the purified wild type CFTase and $CFTase{\Delta}NC$ were 148kDa, 90kDa, respectively.