• 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.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.528-530
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• 2002

We have previously reported recombinant deletion mutant was constructed from cycloinulo-oligosaccharide fructanotransferase(CFTase) gene of Xanthmonas oryzae MGL21. Purification of the mutant protein from E. coli and reation condition for the production of inulo-oligosaccharide(ISO) were studied. The molecular mass of the CFTase deletion mutant protein was estimated to be 90kDa by SDS- Polyacrylamide gel electrophoresis. Optimum reaction pH and temperature were pH6.5 and $40^{\circ}C$, respectively. Under optimal reaction conditions, endo-inulinase activating enzyme was rapidly produced ISO from inulin. Components were DP(degree of polymerization) 3and 4 with trace amount of smaller oligosaccharides.

• Journal of Microbiology and Biotechnology
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• v.12 no.6
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• pp.921-928
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• 2002

Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that 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 ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.18-22
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• 1993

An assay system by using antibody was adopted to screen the endoinulinase producingfungi due to its high specificity toward endoinulinase, To determine whether the affinity-purified rabbit serum, which were generated against the purified endoinulinase, can react only with the endoinulinase, rocket immunoelectrophoresis was performed. The results showed that the serum specifically reacts with endoinulinase but not with exoinulinase and other proteins in the culture media. Using this polyclonal antibody, a strain from 62 fungal colonies was selected and it secreted an endoinulinase in the culture media to the amount comparable to that of Aspergillus ficuum A Tee 16882 known as a high endoinulinase producer.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.278-287
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• 2006

Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cycloinulooligosaccharides (cyclofructan, CF) of ${\beta}-(2{\to}1)$-linked D-fructofuranose as well as hydrolysis of cyclofructan. Sequences analysis indicated that CFTase was divided into five distinct regions containing three repeated sequences (R1, R3, and R4) at the N-terminus and C-terminus. Each domain function was investigated by comparison of wild type CFTase enzyme (CFT148) and deletion mutant proteins (CFT108: R1 and R3 deletion; CFT130: R4 deletion; and CFT88: R1, R3, and R4 deletion) of CFTase. The CFT108 mutant had both CFTase and CF hydrolyzing activity as CFT148 did. CFTase activities and CF hydrolysing activities were disappeared in CFT130 and CFT88 mutants. These results indicated that the C-terminal R4 region of P. polymyxa CFTase is necessary for cyclization and hydrolyzing activity.

• Korean Journal of Medicinal Crop Science
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• v.5 no.3
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• pp.169-176
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• 1997

Gibberellin (GA) has been widely used to break seed dormancy for better stand establishment. The experiment was done to clarify the effect of $GA_3$ (concentration; period) and light quality (red; white; dark) after sowing on seed germination and radicle elongation of Campanulaceae (Platycodon grandiflorum; Codonopsis lanceolata; C. pilosula) to get an information on their field emergence. The germination test was carried out with 12 hours irradiation for 9 days after priming treatment. In the darkness, the mean germination rate of all the species was decreased in the order to P. grandiflorum, C. lanceolata, C. pilosula. Their mean germination rates and radicle lengths were increased with increased concentration to 0.1mM of $GA_3$. Earlier germination rate was higher but later one was less in 4-day $GA_3$ treatment than in 1- or 2-day $GA_3$ treatment. Light treatment. especially red light given after $GA_3$ treatment. eliminated the $GA_3$ treatment effect. Red light done after $GA_3$ treatment nearly blocked the germination of P. grandiflorum and C. pilosula but delayed that of C. pilosula compared to the other light quality treatments having the similar rate. In addition. the radicle elongation of all three species affected by light quality treatment showed the same result as the germination rate.