• Journal of Microbiology and Biotechnology
• /
• 제17권5호
• /
• 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
• /
• 제18권2호
• /
• 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
• /
• 제35권3호
• /
• pp.196-201
• /
• 1992

분질 고구마인 수원 147과 점질인 황미로부터 전분을 분리하고 산가수분해 특성 및 X-ray 회절도를 통해 내부결합력 및 결정강도를 비교하였으며 겔크로마토그래피를 통해 산분해 잔사의 구조적 특성을 조사하였다. 수원 147이 황미보다 산가수분해에 강한 저항성을 보였으며 생전분의 결정화도와 산처리에 따른 전분들의 상대적 결정화도의 증가는 수원 147이 더 크게 나타났다. 산처리에 의하여 ${\beta}$-Amylolysis limit(%)는 점차 증가하였으며 겔크로마토그래피로 용출되는 분자크기가 크게 감소했다. 산분해 잔사는 중합도(DP) 25와 DP 15 부근의 2개의 peak로 분획되었다. Debranching 한 후의 분획은 DP 15부근의 단일 peak로 나타났고 peak의 DP에서 분질과 점질간 차이가 나타났다.

• Journal of Chest Surgery
• /
• 제47권6호
• /
• 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
• /
• 제47권6호
• /
• 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
• /
• 제48권1호
• /
• 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
• /
• 제29권1호
• /
• 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
• /
• 제4권1호
• /
• pp.72-79
• /
• 2007

목 적 : 제3형 당원병은 상염색체 열성으로 유전되는 드문 글리코겐 대사 질환이다. 글리코겐 debranching 효소는 두 가지 효소의 기능을 가지는데, amylo-1,6-glucosidase와 4-alpha-glucanotransferase가 그것이며, 제3형 당원병에서는 글리코겐 debranching 효소의 결핍으로 글리코겐의 불완전한 분해가 초래되며, 다양한 임상 및 생화학적 양상을 보이는 것으로 알려져 있다. 본 연구에서는 3명의 한국인 환자의 임상 및 생화학적 양상을 분석하고, AGL 유전자의 돌연변이 형태를 밝히고자 하였다. 방 법 : 서로 혈연 관계가 없는 3명의 한국인 제3형 당원병 환자를 대상으로, 생화학적, 조직학적, 방사선학적 특징을 포함한 임상 양상을 의무 기록을 통하여 조사하였다. 환자의 말초혈액에서 백혈구를 분리하여 추출한 genomic DNA를 사용하여 직접적 염기 서열 분석법으로 AGL 유전자의 35개 exon 및 exon과 intron의 경계 부분을 조사하여 돌연변이를 조사하였다. 결 과 : 간비대, 경련, 저신장, 고지혈증, 간효소 수치의 증가, creatine kinase 수치의 증가, 경도의 심근증 등 다양한 임상 양상이 관찰되었고, 한 명의 환자는 진행성 간섬유화로 인하여 간이식 수술을 시행 받았다. 생옥수수 전분가루의 복용은 모든 환자에서 정상 혈당을 유지시키고, 생화학적 검사 소견을 개선시키며 정상적인 성장 속도를 보이게 하였다. AGL 유전자 분석 결과 6개의 대립유전자 중 5개에서 돌연변이를 확인할 수 있었으며, 이중에 p.R428K를 제외한 4개의 돌연변이는 이제까지 보고된 적이 없는 새로운 돌연변이(c.1306delA, c.1510-1511insT, c.3416 T>C, c.1735+1 G>T)였다. 결 론 : 제3형 당원병은 임상 증상 및 중등도가 다양한 질환으로, 제1형 당원병의 증상과 유사하여 초기에 감별이 쉽지 않으며, 한국인 환자에서의 AGL 유전자의 돌연변이 양상도 매우 이질적이다.

• 대한유전성대사질환학회지
• /
• 제6권1호
• /
• 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.

• 한국식품과학회지
• /
• 제21권1호
• /
• pp.80-85
• /
• 1989

겉보리와 쌀보리의 전분 및 이들의 구성성분인 아밀로오스와 아밀로펙틴의 분자 구조적 성질을 비교 검토하였다. 쌀보리의 전분 및 아밀로오스와 아밀로펙틴의 ${\beta}-amylosis\;limit(%)$는 각각 56.3, 87 및 52% 이었고, 겉보리 58.6, 77.7 및 57.6%이었다. 쌀보리 아밀로펙틴의 outer chain length${\overline{OCL})$과 inner chain length ${\overline{ICL}}$을 각각 15.8 및 7.8 이었고 겉보리는 각각 17.0 및 7.8이었고, 두 chain 길이의 비는 각각 약 2.1:1 및 2.0:1이었다. 두 시료 전분 모두 pullulanase 로 debranching 한 후의 Sephadex G-75에 의한 용출곡선의 양상은 ${\overline{dp}}55$ 이상의 아밀로오스 성분을 나타내는 분획과 아밀로펙틴 성분으로 볼 수 있는 ${\overline{dp}}35-45$ 및 ${\overline{dp}}10-20$의 bimodal 분포를 갖는 분획 특징을 나타내었다. 또 debranching과 동시에 ${\beta}-amylase$로 처리한 경우는 가수분해 되지 않는 ${\overline{dp}}55$ 이상의 분획성분이 $0.3{\sim}1.5%$ 존재하였다. 아울러 ${\beta}-amylase$로만 가수분해한 경우도 시료차이 없이 ${\overline{dp}}\;55$ 이상의 ${\beta}-limit\;dextrin$ 성분과 ${\overline{dp}}\;10$ 이하의 과당성분으로 분획되는 비슷한 분획양상을 보였으나 분획성분의 분자량 분포사이에는 현저한 시료 차이를 나타내었다.