TY - JOUR
T1 - Cloning and characterization of glycogen-debranching enzyme from hyperthermophilic archaeon Sulfolobus shibatae
AU - Kim, Van Trinth Thi
AU - Ryu, Soo In
AU - Lee, Kyung Ju
AU - Kim, Eun Ju
AU - Lee, Soo Bok
PY - 2007/5
Y1 - 2007/5
N2 - A gene encoding a putative glycogen-debranching enzyme in Sulfolobus shibatae (abbreviated as SSGDE) was cloned and expressed in Escherichia coli. The recombinant enzyme was purified to homogeneity by heat treatment and Ni-NTA affinity chromatography. The recombinant SSGDE was extremely thermostable, with an optimal temperature at 85°C. The enzyme had an optimum pH of 5.5 and was highly stable from pH 4.5 to 6.5. The substrate specificity of SSCIDE suggested that it possesses characteristics of both amylo-1,6-glucosidase and α-1,4-glucanotransferase. SSGDE clearly hydrolyzed pullulan to maltotriose, and 6-O-α-maltosyl-β-cyclodextrin (G2-β-CD) to maltose and β-cyclodextrin. At the same time, SSGDE transferred maltooligosyl residues to the maltooligosaccharides employed, and maltosyl residues to G2-β-CD. The enzyme preferentially hydrolyzed amylopectin, followed in a decreasing order by glycogen, pullulan, and amylose. Therefore, the present results suggest that the glycogen-debranching enzyme from S. shibatae may have industrial application for the efficient debranching and modification of starch to dextrins at a high temperature.
AB - A gene encoding a putative glycogen-debranching enzyme in Sulfolobus shibatae (abbreviated as SSGDE) was cloned and expressed in Escherichia coli. The recombinant enzyme was purified to homogeneity by heat treatment and Ni-NTA affinity chromatography. The recombinant SSGDE was extremely thermostable, with an optimal temperature at 85°C. The enzyme had an optimum pH of 5.5 and was highly stable from pH 4.5 to 6.5. The substrate specificity of SSCIDE suggested that it possesses characteristics of both amylo-1,6-glucosidase and α-1,4-glucanotransferase. SSGDE clearly hydrolyzed pullulan to maltotriose, and 6-O-α-maltosyl-β-cyclodextrin (G2-β-CD) to maltose and β-cyclodextrin. At the same time, SSGDE transferred maltooligosyl residues to the maltooligosaccharides employed, and maltosyl residues to G2-β-CD. The enzyme preferentially hydrolyzed amylopectin, followed in a decreasing order by glycogen, pullulan, and amylose. Therefore, the present results suggest that the glycogen-debranching enzyme from S. shibatae may have industrial application for the efficient debranching and modification of starch to dextrins at a high temperature.
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M3 - Article
C2 - 18051301
AN - SCOPUS:34250184680
VL - 17
SP - 792
EP - 799
JO - Journal of Microbiology and Biotechnology
JF - Journal of Microbiology and Biotechnology
SN - 1017-7825
IS - 5
ER -