• Food Science and Biotechnology
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• v.18 no.2
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• pp.570-573
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• 2009

The debranching enzyme of Nostoc punctiforme (NPDE) is a novel enzyme that catalyzes the hydrolysis of $\alpha$-1,6-glycosidic linkages in starch, followed by the sequential hydrolysis of $\alpha$-1,4-glycosidic linkages. The debranching activity of NPDE is highly specific for branched chains with a degree of polymerization (DP)>8. Moreover, the rate of hydrolysis of $\alpha$-1,4-linkages by NPDE is greatly enhanced for maltooligosaccharides (MOs) with a DP>8. An analysis of reaction mixtures containing various starches revealed the accumulation of maltooctaose (G8) with glucose and maltose. Based on the novel enzymatic properties of NPDE, an MO mixture containing more than 60% G8 with yield of 18 g G8 for 100 g starch was prepared by the reaction of NPDE with soluble starch, followed by ethanol precipitation and gel permeation chromatography (GPC). The yield of the G8-rich mixture was significantly improved by the addition of isoamylase. In summary, a 4-step process for the production of a G8-rich mixture was developed involving the enzymatic hydrolysis of starch by NPDE.

• The Korean Journal of Food And Nutrition
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• v.10 no.1
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• pp.82-86
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• 1997

A Korean traditional sweet rice drink "Sikye" was produced from the raw material of 20% of rice and 4% malt supplemented with 2l of tap water, by incubating the mixture at 6$0^{\circ}C$ for 7 hours. The product was found to contain 11.01% of maltose, 5.31% of isomaltooligosaccharides, 1.75% of maltotriose and 0.28% of glucose. Maltose, maltotriose and isomaltooligosaccharides in Sikye were seperated by ethanol (3 volume) precipitation repeated three times, followed by gel chromatography of Toyopearl HW-40S. 1H-NMR analysis revealed that the products of G2 and G3 size had only $\alpha$-1, 4-glucosidic linkage. but isomaltooligosaccharides showed both signal of $\alpha$-1, 4 and $\alpha$-1, 6-glucosidic linkage with its estimation ratio of 5:1. Isomaltooligosaccharides were hydrolyzed to produce maltooligosaccharide series from maltose to maltohexaose by pullulanase. These results, suggest that isomaltooligosaccharides were constructed by maltohexaose main chain with maltose or maltotriose and maltotetraose side chain.ide chain.

• KSBB Journal
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• v.19 no.4
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• pp.269-273
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• 2004

A simple, fast and reproducible quantitative analysis method for sugar concentration composed in oligosaccharide mixture was developed. Two glass TLC plates were prepared per sample. After dipping one plate into the copper bicinchoninate reagent and the other plate into 5% sulfuric acid solution, both plates were baked in microwave oven until sugar spots were developed or the surface temperature of TLC plate becomes 60 to 70 $^{\circ}C$. The corrective factor values [F value =(the value of total sugar concentration converted as glucose unit/the value of reducing sugar concentration converted as glucose unit)/(polymerization degree of sugar)] of different molecular weight sugars were determined. Within the concentration of 0.25∼1.0 $\mu\textrm{g}$ in each sample loaded, the fructose-F (corrective factor value of fructose) was 0.45, yet for the higher concentration (2.5∼7.5 $\mu\textrm{g}$) fructose-F was 1.0. In case of glucose, in the range of 0.5∼7.5 $\mu\textrm{g}$, glucose-F was same as fructose-F, 1.0. However, as the molecular weight of sugar was increased, the F values were decreased in both maltodextrin and isomaltodextrin oligosaccharides in 0.5∼7.5 $\mu\textrm{g}$ of each sample loaded. Interestingly, F values were equal for the same molecular weight sugars, although the structures were different from each other. Using F value of each sugar, we could determine and compare the exact total sugar concentration of different molecular weight maltooligosaccharide and isomaltooligosaccharide. We also could determine if the unknown sugar was a reducing or non-reducing compound by using optimized TLC with microwave oven method.

• Korean Journal of Microbiology
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• v.36 no.3
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• pp.203-208
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• 2000

A Baczllus circdans KCTC3004 $\alpha$-amylase gene contained in a recombinant plasmid pAL850 was transferred into a new shuttle vector plasmid pALSIlI by ligating linearlzed DNAs of pUC19 and pUB110. B. subtilis RM125 and B. megatenurn ATCC14945 transfonned with pALS111 produced the $\alpha$-amylase substantially Most of the enzyme was produced during the exponential growth period. The maxiinurn activities of the $\alpha$-amylase produced by the Bucillus transformants were compared with that of the B. circulans gene donor strain. The B. subtilis RM125(pALS111) enzyme showed the actlvicy 95 times higher than that of the gene donor cells, followed by the B, nzegaterium ATCC14945(pALSlll) enzyme with activity 34 limes higher than that of the gene donor cells. While E coli secreted about 10% of the produced enzyme, B. subtilis excreted the enzyme inlo the medium wholly and B. megaterirun about 98% ofthe total product. The plasmid pALSI11 was quite stable inB. nzegaterium (92%), inoderately stable in B. subtilis (76%), but was unstable in E. coli (38%). The SDS-PAGE and zymogram of this enzyme produced in E. coli(pALS111), B. subtilis( pALS111) or B. megateril~m (pALS111) indicated a molecular weight of 55,000. The enzymes overproduced in three different host cells hydrolyzed starch to produce mainly maltoaiose and mallooligosaccharides.