• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.123-129
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• 2007

The trehalose $({\alpha}-D-glucopyranosyl-[1,1]-{\alpha}-D-glucopyranose)$ biosynthesis genes MhMTS and MhMTH, encoding a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively, have been cloned from the hyperthermophilic archaebacterium Metallosphaera hakonesis. The ORF of MhMTS is 2,142 bp long, and encodes 713 amino acid residues constituting a 83.8 kDa protein. MhMTH is 1,677 bp long, and encodes 558 amino acid residues constituting a 63.7 kDa protein. The deduced amino acid sequences of MhMTS and MhMTH contain four regions highly conserved for MTSs and three for MTHs that are known to constitute substrate-binding sites of starch-hydrolyzing enzymes. Recombinant proteins obtained by expressing the MhMTS and MhMTH genes in E. coli catalyzed a sequential reaction converting maltooligosaccharides to produce trehalose. Optimum pH of the MhMTS/MhMTH enzyme reaction was around 5.0 and optimum temperature was around 70 C. Trehalose-producing activity of the MhMTS/ MhMTH was notably stable, retaining 80% of the activity after preincubation of the enzyme mixture at $70^{\circ}C$ for 48 h, but was gradually abolished by incubating at above $85^{\circ}C$. Addition of thermostable $4-{\alpha}-glucanotransferase$ increased the yield of trehalose production from maltopentaose by 10%. The substrate specificity of the MhMTS/MhMTH-catalyzed reaction was extended to soluble starch, the most abundant maltodextrin in nature.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1426-1431
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• 1998

The research was undertaken to characterize enzymatic properties of Streptomyces albus amylase expressed in recombinant Bacillus subtilis. Molecular weight and pI of the purified enzyme were estimated to be 50 kD by SDS-PAGE and 4.3 by isoelectric focusing. The optimum temperature and optimum pH were $45^{\circ}C$ and 6.0, respectively. D-and Z-value were estimated to measure thermostability of the purified enzyme. The Z-value was estimated $17.7^{\circ}C$, which is lower than typical amylase. Maltotetraose was produced as a major component from soluble starch in the early state of reaction but gradually degraded to maltose. Thin layer chromatography was also performed to analyze the reaction products. The parameters involved in Michaelis-Menten enzyme kinetics were found to be the maximum velocity of 0.37 mM/min and the Michaelis constant of 0.13%, respectively.

• Journal of Life Science
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• v.21 no.2
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• pp.221-226
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• 2011

The gene encoding 4-$\alpha$-glucanotransferase (mgtA) from Thermotoga neapolitana was cloned and expressed in Escherichia coli in order to investigate whether this enzyme was capable of producing cycloamylose for industrial applications. MgtA was purified to homogeneity by HiTrap Q HP and Sephacryl S-200 HR column chromatographies. The size of the enzyme as determined by SDS-PAGE was about 52 kDa, which was in good agreement with its deduced molecular mass of 51.9 kDa. The optimal temperature and pH for the activity of the 4-$\alpha$-glucanotransferase was found to be $85^{\circ}C$ and 6.5, respectively. The enzyme hydrolyzed the 1,4-$\alpha$-glucosidic bonds in oligomeric 1,4-$\alpha$-glucans and transferred oligosaccharides (maltotriose being the shortest one) to acceptor maltodextrins. However, the enzymes had no activity against pullulan, glycogen, and other di- or trioligosaccharides with rare types of $\alpha$-bond. MgtA is distinguished from 4-$\alpha$-glucanotransferase from Thermotoga maritima in that it can convert maltotriose into maltooligosaccharides. The treatment of glucoamylase after the reaction of MgtA with maltotriose, maltotetraose, maltopentaose, or maltohexaose as sole substrate revealed that MgtA yielded linear maltooligosaccharides instead of cycloamylose.

• Food Science and Preservation
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• v.15 no.1
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• pp.99-104
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• 2008

To utilize non-heat treated alcoholic by-products of brown rice (Goami) as food sources, the quality characteristics changes according to the treatment conditions of cellulase were evaluated. Results showed that the increase of hydrolysis temperature correspondingly increased the soluble solids and total sugar amounts in the by-products of Goami, and total dietary fiber amount was found to be around 0.67% Reducing sugar concentration was the highest at the hydrolysis temperature of $70^{\circ}C$. Maltooligosaccharides amounts were detected to be the highest at the hydrolysis temperature of $80^{\circ}C$ and were also, maltopentose and maltopentose were found. In the soluble solid, total dietary fiber, reducing sugar and total sugar according to the cellulase concentration, the content of hydrolysates with enzyme were higher than control, and the content of hydrolysates with enzyme was similar (6.30 and 0.69% 3,600 and 5,500 mg% respectively). The content of maltooligosaccharides was increased with the increase of enzyme concentration, and the content was similar at more than 0.6%(w/w) of enzyme concentration. The soluble solids and total dietary fiber by hydrolysis time were found to be 6.25% and 0.70%, respectively at more than 60 min. of hydrolysis. The content of reducing sugar, total sugar and maltooligosaccharides were increased with the increase of hydrolysis time, and the content was similar at more than 120min. of hydrolysis (3,800, 5,680 and 1,950 mg% respectively). Based upon these results, the byproducts of Goami are expected to be valuable as various food sources showing the highest dietary fiber and maltooligosaccharides contents by the hydrolysis at $80^{\circ}C$ for 120 min. with the addition of 0.6%(w/w) of cellulase.