• Journal of Life Science
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• v.17 no.9 s.89
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• pp.1182-1190
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• 2007

The gene encoding for isoamylase of the Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 was cloned and expressed into Escherichia coli $DH5{\alpha}$. Isoamylase catalyzes the hydrolysis of ${\alpha}-1,6-glycosidic$ linkages specifically in amylopectin, glycogen, and derived oligosaccharides, while the enzyme did not hydrolyze ${\alpha}-1,4-glycosidic$ linkages of amylose. The isoamylase gene (glgX) had an open reading frame of 1,977 bp encoding 658 amino acid residues with a calculated molecular weight of 74,188 Da. The molecular weight of the enzyme was also estimated to be 74 kDa by activity staining of a SDS-PA gel. The mature GlgX had a calculated pI of 4.91. Isoamylase from Pcc LY34 had 70% amino acid identity with isoamylase from Pectobacterium chrysanthemi and contained the four regions conserved among all amylolytic enzymes. The isoamylase was optimally active at pH 7.0 and $40^{\circ}C$. GlgX was $Ca^{2+}-dependent$. The changes of Asp-335, Glu-370, and Asp-442 into Ala, respectively, using site-directed mutagenesis techniques showed that three residues are essential to isolamyalse (GlgX) activity. The sequences around those residues were highly conserved in isoamylase of different origins and GlgX of the glg operon in glycongen biosynthesis.

• Applied Biological Chemistry
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• v.13 no.3
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• pp.181-186
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• 1970

1) Conditions for the hydrolysis of starch by bacterial liquefying amylase (BLA), saccharifying amylase (BSA) and isoamylase were investigated. Out of four syrups prepared by different combinations of these enzymes, those made by BLA followed by BSA and/or isoamylase were comparable to sucrose syrup in canning of orange segments. 2) Two branched maltooligosaccharides were isolated from the hydrolyzate of starch by BLA and BSA, and their structures were tentatively identified as pentaose and hexaose having an ${\alpha}-1$, 6-linkage at the branching point.

• The Korean Journal of Food And Nutrition
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• v.9 no.1
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• pp.45-51
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• 1996

Korean traditional Sikhye is made from rice and malt, and It's main product Is maltose. However commercial Sikhye differs from traditional Sikhye because it's main component is sucrose. Sikhye industry faces many problems such as contamination of malt with microorganisms, low amylase activity of malt and technical difficulties. There is no commercial Sikhye which is only using rice and malt by these reasons. To produce the traditional Sikhye free from these problems, it is necessary to restrict the microorganisms of malt and to standardize the amylase activity of malt. In addition, the Introduction of effective control and sanitaric process is required. In Sikhye production. if $\beta$-amylase and isoamylase or pullulanase were added, starch could be saccharified 100% as maltose. Accordingly, this method brings us the low cost of Sikhye.

• The Korean Journal of Pharmacology
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• v.16 no.2 s.27
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• pp.15-24
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• 1980

[${\alpha}$]-Amylase catalyses the hydrolysis of starch, glycogen, and related poly- and oligosac-charide by random cleavage of ${\alpha}$-D-(l-4) glucan linkage. In man large amounts of amylase are secreted into the digestive tract by the salivary and exocrine pancreatic gland, minimal amount being produced also in other tissues. It has been known that ${\alpha}$-amylase exists in multiple molecular forms, isoenzyme which can be separated from each other because of difference in their physicochemical properties. By using various methods, several groups of investigator have separated the many isoenzyme in serum, saliva and pancreatic juice. Furthermore, changes of the normal serum isoenzyme pattern is diagnostically useful even when the total serum enzyme activity is noninformative, such as the clinical use of isoenzyme of serum lactate dehydrogenase. Procarboxypeptidase-A which is one of the pancreatic enzymes is also present as isoenzymes. Four forms of procarboxypeptidase-A haye been found in the bovine enzyme and three forms of the porcine enzyme. In human pancreatic juice four forms of procarboxypeptidase-A isoenzyme were found by isoelectric focusing method. Recently, the so-called isoamylase analysis was developed for the diagnostic use of amylase in pancreatic diseases. In alcohotic patients, the serum concentration of pancreatic isoamylase is subnormal and this lowered activity provides strong evidence for pancreatic exocrine insufficiency. The purpose of this study was to elucidate the variations of the isoenzyme of amylase and procarboxypeptidase-A in serum, saliva and pancreatic juice of the experimental animals. The results are as follow. 1) Three main forms of isoenzyme of amylase by isoelectric focusing were found in pancreatic juice of normal rabbit. However, many new bands were appeared in the pancreatic juice of cholic acid administered animal intravenously while the infusion of cholic acid or elastase into pancreatic duct produced the decrease of number of the fractions on the isoelectric focusing. In the case of serum isoenzyme from normal animal, two major and a few minor isoamylases were observed. By injecting alcohol intravenousely the fractions of serum isoamylase were significantly decreased and in contrary to the pattern in the pancreatic juice the infusion of cholic acid or elastase into pancreatic duct exhitited a significant decrease of the isoenzyme of amylase fractions. In saliva from normal animal three main isoamylase were produced of the administration of alcohol. 2) In the case of procarboxypeptidase-A isoenzyme, two major fractions which have isoelectric point at 6.2 and 6.4 and other two minor bands were observed in the pancreatic juice of normal rabbit. By the treatment of the juice with trypsin, only one band was produced on the isoelectric focusing. No procarboxypeptidase was appeared on the electrofocusing by the infusion of cholic acid or phospholipase A into the pancreatic duct of rabbit. However, a single major fraction of procarboxypeptidase-A was appeared at 3 hr after simple ligation of the pancreatic duct. No significant changes were observed in the juice of the alcohol or cholic acid administered group.

• The Korean Journal of Food And Nutrition
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• v.9 no.1
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• pp.92-98
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• 1996

Pectic materials, which are widely spread in the plant cell wall as plant carbohydrates, plays a great role in food Industry that acts as a softening agent of fruits and vegetables, and gel forming agents. To study physiochemical properties and industrial applications of pectic enzymes that hydrolyzes pectin, classification, assay method and Industrial application are reviewed based on previous results.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.143-150
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• 1998

Many enzymes catalyze a primary reaction and/or secondary reaction. Dextransucrase usually synthesize dextran from sucrose as a primary reaction. The secondary reaction of dextransucrase is the transfer of glucose from sucrose to carbohydrate accepters. We have reacted dextransucrase from Leuconostoc mesenteroides B-742CB with sucrose and starches; granule or gelatinized starches, and Small or Potato starches. The yield of modified starch was ranged from 46% to 72%(s.d.<${pm}$5%) of theoretical depends on various reaction conditions. Modified products were more resistant against the hydrolysis of ${alpha}$-amylase, isoamylase, pullulanase and endo-dextranase than those of native starch. Based on the reactions from enzyme hydrolysis and methylation followed by acid hydrolysis modification of granule starch was more efficient than the modification of gelatinized starch. After modification of granule starch with dextransucrase, there produced a soluble modified starch. After modification the starch granules were fractionated to small size. The positions of glucose substitution of the modified products were determined by methylation followed by acid hydrolysis and analyzed by TLC. The products were modified by the addition of glucose to the position of C3, C4 and C6 free hydroxyl group of glucose residues in the starch.

• Korean Journal of Food Science and Technology
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• v.53 no.5
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• pp.570-577
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• 2021

This study elucidated the biological activities and structural characteristics of polysaccharides isolated from ginseng leaves fermented using Cordyceps sinensis (GLF). GLF comprised at least 18 glycosyl linkages, including 4-linked glucose residues (24.0%). To characterize the neutral polysaccharides in GLF, it was further fractionated by anion exchange chromatography, and the unabsorbed fraction (GLF1) was isolated. Peritoneal macrophages stimulated with GLF1 produced various cytokines in a dose-dependent manner. The properties and activities of the four subfractions (PHI, PHIA1-PHIA3) obtained after sequential enzymatic digestion were examined. PHI and PHIA3 primarily comprised glucose, whereas PHI exhibited an iodine-color reaction. Furthermore, the PHIA1-3 fractions indicated that cytokine production was completely inhibited. These results suggest at the immune activities of GLF1 may be due to the α-(1→4)-glucan branched at the C(O)6 position, which was produced by C. sinensis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.2
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• pp.255-260
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• 1996

증편반죽의 발효에 따른 해면상의 조직(망상구조)형성능의 mechanism을 조사하기 위하여 반죽의 구성성분의 경시적 물성 및 효소(diastase 및 protease) 활성의 변화, 구성 전분 분획의 glucose chain length 변화 및 1% SDS에 의해서 용출되는 단백질 분획의 변화에 대해서 검토하였다. 발효가 진행됨에 따라 증편반죽의 pH는 감소한 반면 점성 및 부피는 발효경과 10시간가지 증가하다가 그 이후로는 약간 감소하는 경향을 보였다. 발효의 진행과 더불어 diastase의 활성은 증가하였으며, 쌀전분 분획 중 amylose의 함량은 약간 감소하였다. 그리고 전분분자의 $\alpha-1,6-glucoside$ 결합을 isoamylase(debranching enzyme)로 가수분해시킨 후 Sephadex G-75를 이용한 chromatogram 분석에 의하면 쌀가루 전분 분획과 증편반죽 전분 분획의 glucose chain length 분포는 거의 유사하지만 발효가 진행됨에 따라 중합도가 낮은 부분이 우선적으로 diastase의 작용을 받은 것이라고 생각되어진다. 한편, 발효가 진행된에 따라 protease의 활성은 증가하고 있었음에도 불구하고 증편반죽의 당에 의한 단백질 분획의 고분자화가 Suprose CL-12 column chromatogram 상에서 관찰되었는데, 이것은 아마도 발효과정 중 증편반죽에 공존하는 미생물들의 발효산물인 당질(gum질)을 매개로 한 단백질 분자의 회합에 의한 결과라고 생각되어진다. 따라서 이러한 결과를 미루어 볼 때 증편반죽의 망상구조 형성능은 발효과정 중 일어나는 당과 단백질간의 상호작용에 의한 결과라 생각되어진다.

• Korean Journal of Food Science and Technology
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• v.33 no.1
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• pp.50-54
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• 2001

The amylose content, based on iodine blue value, of eight rice cultivars decreased in order of Nampungbyeo>Whachungbyeo>Punchilmi>Nampung CB243>Whachung du-1>Nampung EM90>Whachungchalbyeo>shr. The amylopectin chain length distribution was obtained by enzyme treatments followed by high-performance size-exclusion chromatographic separation. Chain length distribution profiles of the isoamylase-debranched starches showed distinct patterns according to cultivars. Based on the sensory evaluation result of the bread prepared from gluten and rice flours of eight rice cultivars, chewiness of the product was related with the presence of amylose while the short-chain amylopectin fraction was contributed to the texture and overall quality.

• Journal of Plant Biotechnology
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• v.36 no.3
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• pp.207-213
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• 2009

Starch serves not only as an energy source for plants, animals, and humans but also as an environmentally friendly alternative for fossil fuels. Progress in understanding of starch biosynthesis, and the isolation of many genes involved in this process have enabled the genetic modification of crops in a rational manner to produce novel starches with improved functionality. Starch is composed of two glucose polymers, amylose and amylopectin. The amylose and amylopectin ratio in starch affects its physical and physicochemical properties. Alteration in starch structure can be achieved by modifying genes encoding the enzymes responsible for starch biosynthesis and starch hydrolysis. Here, we describe recent findings concerning the starch modification in sweetpotato. Sweetpotato [Ipomoea batatas (L.) Lam] ranks seventh in annual production among food crops in the world as an important starch source. To develop transgenic sweetpotato plants with modifying starch composition, we constructed transformation vectors overexpressing granule bound starch synthase I and inhibiting amylopectin synthesis genes such as starch branching enzyme and isoamylase under the control of 35S promoter, respectively. Transformation of sweetpotato (cv. Yulmi) is in progress.