• Journal of Microbiology and Biotechnology
• /
• v.17 no.5
• /
• pp.792-799
• /
• 2007

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 $85^{\circ}C$. The enzyme had an optimum pH of 5.5 and was highly stable from pH 4.5 to 6.5. The substrate specificity of SSGDE suggested that it possesses characteristics of both amylo-1,6-glucosidase and $\alpha$-1,4-glucanotransferase. SSGDE clearly hydrolyzed pullulan to maltotriose, and $6-O-\alpha-maltosyl-\beta-cyclodextrin(G2-\beta-CD)$ to maltose and $\beta$-cyclodextrin. At the same time, SSGDE transferred maltooligosyl residues to the maltooligosaccharides employed, and maltosyl residues to $G2-\beta-CD$. 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.

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

• Applied Biological Chemistry
• /
• v.35 no.3
• /
• pp.196-201
• /
• 1992

Characteristics on lintnerization of dry type (Suwon 147) and moist type (Hwangmi) sweet potato starches were investigated. Chain distribution of lintnerized starches was also studied by debranching with pullulanase. Hydrolytic patterns of two starches showed two distinct stages and hydrolysis extents of Suwon 147 starch were lower than those of Hwangmi starch. The relative crystallinities of Suwon 147 starches were higher than those of Hwangmi starch. The elution profiles of lintnerized starches were composed of two peaks about degree of polymerization (DP) 25 and DP 15. The elution profiles of debranched samples showed only one peak about DP 15 and peak DP of Suwon 147 lintnerized starch was higher than that of Hwangmi.

• Journal of Chest Surgery
• /
• v.47 no.6
• /
• pp.548-551
• /
• 2014

Type II chronic dissecting thoracoabdominal aortic aneurysms are a surgically challenging disease. The conventional thoracoabdominal aortic aneurysm repair technique using cardiopulmonary bypass is a high-risk procedure. However, a recently developed endovascular technique may be an alternative treatment for the disease, but faces the obstacle of lesional restriction. This new technique uses a hybrid strategy to overcome the limits of endovascular thoracoabdominal aortic aneurysm repair. Herein, we report on a successful outcome after performing the hybrid visceral debranching procedure.

• Journal of Chest Surgery
• /
• v.47 no.6
• /
• pp.552-555
• /
• 2014

Thoracic endovascular aortic repair has become a widespread alternative treatment option for thoracic aortic aneurysm. The debranching of arch vessels may be required to provide an acceptable landing zone for an endovascular stent graft. We report a case where the bypass graft used in the thoracic endovascular aortic repair procedure compressed the left internal jugular vein, causing acute thrombotic occlusion.

• Journal of Chest Surgery
• /
• v.48 no.1
• /
• pp.74-77
• /
• 2015

Chylothorax is a rare postoperative complication of a thoracic surgical procedure. Here, we report a case of chylothorax after thoracic endovascular aortic repair with debranching for the distal arch aneurysm of the aorta. First, the patient was treated by a medical method (nil per os, fat-free diet, and octreotide), but this method failed. The patient strongly refused surgical treatment. Therefore, we tried to occlude the thoracic duct by lymphangiography Lipiodol, and this line of treatment was successful.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.1
• /
• pp.37-43
• /
• 2019

The gene encoding an ${\alpha}-{\text\tiny{L}}-arabinofuranosidase$ (BvAF) GH51 from Bacillus velezensis FZB42 was cloned and expressed in Escherichia coli. The corresponding open reading frame consists of 1,491 nucleotides which encode 496 amino acids with the molecular mass of 56.9 kDa. BvAF showed the highest activity against sugar beet (branched) arabinan in 50 mM sodium acetate buffer (pH 6.0) at $45^{\circ}C$. However, it could hardly hydrolyze debranched arabinan and arabinoxylans. The time-course hydrolyses of branched arabinan and arabinooligosaccharides (AOS) revealed that BvAF is a unique exo-hydrolase producing exclusively ${\text\tiny{L}}-arabinose$. BvAF could cleave ${\alpha}-(1,2)-$ and/or ${\alpha}-(1,3)-{\text\tiny{L}}-arabinofuranosidic$ linkages of the branched substrates to produce the debranched forms of arabinan and AOS. Although the excessive amount of BvAF could liberate ${\text\tiny{L}}-arabinose$ from linear AOS, it was extremely lower than that on branched AOS. In conclusion, BvAF is the arabinan-specific exo-acting ${\alpha}-{\text\tiny{L}}-arabinofuranosidase$ possessing high debranching activity towards ${\alpha}-(1,2)-$ and/or ${\alpha}-(1,3)-linked$ branches of arabinan, which can facilitate the successive degradation of arabinan by $endo-{\alpha}-(1,5)-{\text\tiny{L}}-arabinanase$.

• Journal of Genetic Medicine
• /
• v.4 no.1
• /
• pp.72-79
• /
• 2007

Purpose : Glycogen storage disease type III (GSD-III) is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme, amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), is responsible for the debranching of the glycogen molecule during catabolism. The disease shows clinical and biochemical heterogeneity, reflecting genotype-phenotype heterogeneity among different patients. In this study, we aim at analyzing mutations of the AGL gene in three unrelated Korean GSD-III patients, and characterizing their clinical and laboratory findings. Methods : We characterized the clinical features of three unrelated Korean GSD-III patients by biochemical, histological and imaging studies. The 35 exons and part of exon-intron boundaries of AGL were analyzed by direct sequencing using genomic DNA extracted from the peripheral leukocytes of patients. Results : Diverse clinical features were observed in these patients including hepatomegaly (all patients), seizures (patient 2), grow th failure (patients 1 and 2), hyperlipidemia (patients 1 and 3), raised transaminase and creatine kinase concentrations (all patients), and mild cardiomyopathy (patient 2). Liver transplantation w as performed in patient 2 due to progressive hepatic fibrosis. A dministration of uncooked corn starch maintained normoglycemia and improved biochemical and growth profiles. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 had c.1510_1511insT (p.Y 504L fsX 10), and patient 3 had c.3416 T >C (p.L 1139P) and c.1735+1 G>T (p.Y 538_R578delfsX 4) mutations. A part from the p.R428K mutation, the 4 other substitutions identified w ere nov el. Conclusion : GSD-III patients display variable phenotypic characteristics resembling those of GSD-Ia. Molecular defects in the AGL gene of Korean GSD-III patients are genetically heterogeneous.

• Journal of The Korean Society of Inherited Metabolic disease
• /
• v.6 no.1
• /
• pp.15-23
• /
• 2006

Purpose: Glycogen storage disease type III (GSD-III), is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme is amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), which is responsible for the debranching of the glycogen molecule during catabolism. The disease has been demonstrated to show clinical and biochemical heterogeneity, reflecting the genotype-phenotype heterogeneity among different patients. In this study, we analyzed mutations of the AGL gene in three unrelated Korean GSD-III patients and discussed their clinical and laboratory implications. Methods: We studied three GSD-III patients and the clinical features were characterized. Sequence analysis of 35exons and part exon-intron boundaries of the AGLgene in patients were carried out by direct DNA sequencing method using genomic DNA isolated from patients' peripheral leukocytes. Results: The clinical features included hepatomegaly (in all patients), seizures (in patient 2), growth failure (in patients 1), hyperlipidemia (in patients 1 and 3), raised transaminases and creatinine kinase concentrations (in all patients) and mild EKG abnormalities (in patients 2). Liver transplantation was performed in patient 2due to progressive hepatic fibrosis. Administration of raw-corn-starch could maintain normoglycemia and improve the condition. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 with c.1510_1511insT (p.Y504LfsX10), and patient 3 with c.3416 T>C (p.L1139P) and c.l735+1 G>T (Y538_R578delfsX4) mutations. Except R428K mutation, 4 other mutations identified in3 patients were novel. Conclusion: GSD-III patients have variable phenotypic characteristics resembling GSD-Ia. The molecular defects in the AGL gene of Korean GSD-III patients were genetically heterogeneous.

• Korean Journal of Food Science and Technology
• /
• v.21 no.1
• /
• pp.80-85
• /
• 1989

The structural properties of covered and naked barley starches and those components were investigated. ${\beta}-amylosis\;limit(%)$ of covered and naked barley starches were 58.6 and 56.3%, respectively and those of their amyloses and amylopectins were 87.0, 77.7, 57.6 and 52.0%, respectively. The ratios of outer chain length ${\overline{OCL}}$ and inner chain length${\overline{ICL}}$ for amylopectins of covered and naked barley were about 2.2:1 and 2.0:1, respectively. The elution curves by Sephadex G-75 after debranching starches with pullulanase were similar patterns for two starches and yielded two peaks consisting of amylose component${\overline{dp}}>55$ and bimodal amylopectin components of ${\overline{dp}_(35-45)$ and ${\overline{dp}}(10-20)$ hydrolysates. Also, hydrolysis products by sequential hydrolysis with pullulanase and ${\beta}-amylase$ contained 0.03-0.5% non-hydrolyzed peak components of ${\overline{dp}}>55$. The elution profiles of ${\beta}-amylase$ hydrolysates yielded two peaks consisting of the inner components${\overline{dp}}>55$ and the outer chain components of amylopectin${\overline{dp}}>10$ regardless of samples. However, ${\overline{dp}}$ distributions of ${\overline{dp}>55}$ hydrolysates exhibited the significant differences due to the varieties.