• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.31-36
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• 2001

Paenibacillus sp. JB-13 producing the cyclodextrin glucan-otransferase(CGTase) [EC 2.4.1.19] that glucosylated ascorbic acid(AA) at the C-2 position was isolated form soil and the optimal conditions for the production of 2-O-$\alpha$-D- Glucopyranosl L-Ascorbic acid(AA-2G) with CGTase were investigated. CGTase produced AA-2G efficiently using dextrin as a substrate and AA as an aceptor. Several AA-2-oilgosaccharides(AA-2Gs) were also produced in this reaction mixture, and these were efficiently hydro-lyzed to AA-2G and glucose by the treatment with glucoamylase. The optimal temperature for AA-2G production was $37^{\circ}C$ and the optimal pH was around 6.5. CGTase also utilized $\alpha$-,$\beta$-,${\gamma}$-CDs, soluble starch, com statch, dia-static solution from rice and diastatic solution from malt as substrate, but not glucose. The reaction mixture for the maximal production of AA-2G was following; 15% total substrate concentration, 2,500 units/ml of CGTase and a mixing ration of 3:2(g of AA: g of dextrin). Under this condition, 56 mM of AA-2G ,which corresponded to 12.4% yield based on AA. was produced after incubation for 44 hrs at $37^{\circ}C$ and pH 6.5.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.74-79
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• 2008

The purpose of this study is to clone cgt gene from Paenibacillus sp. JB-13 and to overexpress the protein in E. coli. For this purpose, the cgt gene was amplified from Paenibacillus sp. JB-13 genomic DNA by PCR using degenerate oligonucleotide primers. The sequence analysis results showed that the cgt gene from Paenibacillus sp. JB-13 has 98% homology with the cgt gene of Bacillus sp. To overexpress the protein, the cgt gene was cloned into pEXP7 expression vector and transformed into E. coli. The production of CGTase by recombinant E. coli was optimized under following conditions: 0.5% glucose, 3.0% polypeptone, 0.3% $K_2HPO_4$, 0.5% NaCl, and 7.0 of initial pH, 2.0% of inoculum, $37^{\circ}C$ of culture temperature for 14 hr. And the optimal agitation was found at 0.1 vvm. The synthesis of 2-O-${\alpha}$-D-Glucopyranosyl L-Ascorbic acid (AA-2G) using the CGTase expressed in E. coli was identified as AA-2G by HPLC and HPLC confirmed that treating AA-2G made by cloned CGTase with ${\alpha}$-glucosidase substantially produced AA and glucose.

• Journal of Food Hygiene and Safety
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• v.35 no.1
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• pp.83-92
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• 2020

This study investigated the optimal conditions of enzymatic reaction for production of isomaltooligosaccharides (IMO) using rice flour. To manufacture IMO, commercial enzymes (Termamyl 2X, Maltogenase L, Promozyme D2, Fungamyl 800L and Transglucosidase L) were used. The sugar composition and amount of IMO were examined by HPLC with charged aerosol detector (HPLC-CAD) in each manufacturing process. Liquefaction reaction was performed according to different Termamyl 2X concentrations (0.025%, 0.05%, 0.075%, 0.1%) and reaction times (1 h, 2 h). As a result, the reducing sugar content was the highest at 138.26 g/L when 0.075% Termamyl 2X was added for 2 hours. In order to optimize simultaneous saccharification and transglucosylation, experiments on enzyme selection, enzyme concentration and enzyme reaction time were conducted. Reaction with 0.0015% Maltogenase L, 0.05-0.1% Promozyme D2 and 0.1% Tansglucosidase L was effective in decreasing glucose content and increasing content of IMO with a high degree of polymerization. A change in sugar content was observed every 6 hours to determine the optimal reaction time, and the highest IMO was produced after 36 hours of reaction (75.36 g/L). The IMO prepared under optimal conditions showed isomaltose, 35.11 g/L; panose, 11.97 g/L; isomaltotriose, 19.95 g/L; isomaltotetraose, 7.46 g/L; isomaltopentaose, 1.05 g/L at 18 brix and the ratio of IMO in the total sugar was 56.37%.