• Title/Summary/Keyword: isozyme

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Further induction of amylase producing mutants from a highly proteolytic mutant strain of asppergillus flavus (돌연변이에 의한 Aspergillus flavus의 아밀라아제 생성능의 개량)

  • 이영록;고상균;김봉수
    • Korean Journal of Microbiology
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    • v.18 no.4
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    • pp.161-171
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    • 1980
  • A mutant strain having increased productivity of both enzymes, protease and amylase, was obtained from A. flavus KU 153, isolatd from South Korea for its high protease production by successive ultra-violet light irradiation, Two glucoamylases from the mutant strain selected were purified from wheat branculture by successive salting out, followed by dialysis and column chromatography, and their characteristics were compared with those of the wild strain. Glucoamylase production of the mutant selected was increased about 3.3 times compared with the wild strain, and 2.1 times compared with the parental strain, ${\alpha}-amylase$ activity of the mutant selected was about 2 times hugher than that of the wild strain or the parental strain. Protease and cellulase productivities of the muant selected were all alike compared with those of the highly proteolytic mutant, the parental strain. Therefore, it was considered that the back mutation on the protease production did not occurred in the formation process of the glucoamylase producing mutant. Total activities of glucoamylase I and II from the mutant selected were 2.86 and 3.65 times higher compared with those from the wild strain, respectively. Considering the optimal pH-thermal stability and Km-Vmax value of glucoamylase I and II from both strains, wild and mutant, it was deduced that the characteristics of glucoamylase I and II from the wild strain did not altered during the mutation process. Therefore, it was concluded that the selected mutant did not induce the formation of another glucoamylase isozyme, or the changes in the characteristics of the glucoamylase, but induce the productivity of the same glucoamylase I and II by the action of regulatory gene.

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The nervous system of Fibricola seoulensis by acetylcholinesterase histochemistry (Acetylcholinesterase 조직화학법으로 관찰한 Fibricola seoulensis 신경계의 분포)

  • 전은우;김창환
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.321-330
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    • 1993
  • F. seoulensis were obtained from artificially infected albino rats at 3, 4, 5, 6, 7 days after infection. The worms and metacercariae were washed in physiological saline solution, and fled with 10% neutral formalin. The acetylcholinesterase (AchE) stained by one histochemistry using acetylthiocholine iodide as substrate. Eserine, ism-OMPA and BW284C51 were used as inhibitors of AchE. The nervous system consists of three pairs longitudinal nerve trunks interconnected with excretory plexus in posterior half, and phinmc and oral sucker in anterior half of metacercariae and adults. The longitudinal nerve trunks are interconnected with transverse commissures and numerous circular commissures. Considerable numbers of circular commissures are interconnected with longitudinal nerve trunks tying on the surface of the worms. At each stage of juvenile worms, AchE and nonspecific cholinesterase activites were observed in the oral sucker, ventral sucker, pharynx and nerve system. Isoxymes of AchE in f seoderuts were separated into the two bands, 69 kDa and 132 kDa. The major band was 69 kDa.

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Classification of Sweet Potato Varieties Based on Esterase Isozymes and Protein Patterns (Esterase 동위효소 및 단백질 패턴에 의한 고구마 품종 분류)

  • Kim, Deog-Su;Oh, Sung-Kun;Chin, Moon-Sup;Ryu, Jeom-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.3
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    • pp.332-339
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    • 1996
  • Electrophoretic method was utilized to classify 100 varieties of sweet potato germplasm maintained at the National Crop Experiment Station of Korea in 1993. The esterase isozyme patterns in the leaves were classified into 14 different types. Type Ⅸ included the most of the varieties (46) tested and Ⅶ, I, III, Ⅷ, II and V types of all included 47 varieties in order. The other 7 varieties had different band pattern with each other. Type I having many kind of band pattern included Shinyulmi, Beniastma and High starch which had the dry type of tuberous roots varieties. The esterase isozymes pattern in the tuberous roots were classified with 18 kinds of types. The C type included 22 varieties and B, K, A, E, I and N in order. The proteins pattern in the tuberous roots were classified with 7 kinds of types. I type included 36 varieties, and IV type included 27 varieties and II, III, Ⅶ and Ⅵ types in order.

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Raw Inonotus obliquus polysaccharide counteracts Alzheimer's disease in a transgenic mouse model by activating the ubiquitin-proteosome system

  • Shumin Wang;Kaiye Dong;Ji Zhang;Chaochao Chen;Hongyan Shuai;Xin Yu
    • Nutrition Research and Practice
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    • v.17 no.6
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    • pp.1128-1142
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    • 2023
  • BACKGROUND/OBJECTIVES: Inonotus obliquus has been used as antidiabetic herb around the world, especially in the Russian and Scandinavian countries. Diabetes is widely believed to be a key factor in Alzheimer's disease (AD), which is widely considered to be type III diabetes. To investigate whether I. obliquus can also ameliorate AD, it would be interesting to identify new clues for AD treatment. We tested the anti-AD effects of raw Inonotus obliquus polysaccharide (IOP) in a mouse model of AD (3×Tg-AD transgenic mice). MATERIALS/METHODS: SPF-grade 3×Tg-AD mice were randomly divided into three groups (Control, Metformin, and raw IOP groups, n = 5 per group). β-Amyloid deposition in the brain was analyzed using immunohistochemistry for AD characterization. Gene and protein expression of pertinent factors of the ubiquitin-proteasome system (UPS) was determined using real-time quantitative polymerase chain reaction and Western blotting. RESULTS: Raw IOP significantly reduced the accumulation of amyloid aggregates and facilitated UPS activity, resulting in a significant reduction in AD-related symptoms in an AD mouse model. The presence of raw IOP significantly enhanced the expression of ubiquitin, E1, and Parkin (E3) at both the mRNA and protein levels in the mouse hippocampus. The mRNA level of ubiquitin carboxyl-terminal hydrolase isozyme L1, a key factor involved in UPS activation, also increased by approximately 50%. CONCLUSIONS: Raw IOP could contribute to AD amelioration via the UPS pathway, which could be considered as a new potential strategy for AD treatment, although we could not exclude other mechanisms involved in counteracting AD processing.

Effects of Chicory Inulin and Oligosaccharides on Lipid Metabolism in Rats Fed a High-Cholesterol Diet (고콜레스테롤 식이 섭취 흰쥐에서 치커리 이눌린과 올리고당이 지질대사에 미치는 영향)

  • 성혜영;정현진;최영선;조성희;윤종원
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.2
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    • pp.305-310
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    • 2004
  • The present study was aimed at investigating effects of chicory inulin and three kinds of oligosaccharides on lipid metabolism in rats fed a high-cholesterol diet. Nine Sprague-Dawley male rats weighing, about 190g were given one of five experimental diets, which were basal cholesterol diet (Control) isomaltooligosaccharide diet (IMO), Iructooligosaccharide diet (FO), chicory inulooligosaccharide diet (CIO) and chicory inulin diet (CI) for 5 weeks. In the oligosaccharide and inulin diets, 6% was added at the expense of sucrose. Rats were pair-fed to the intake of FO group which consumed the least amount, and their feces were collected during the last 4 days. Body weight gain was lower in Fo and CI groups compared with the Control group. Plasma glucose levels of FO and CIO groups were lower and plasma triglyceride concentrations of FO, CIO, and CI groups were lower than those of IMO group. Plasma cholesterol concentration did not differ among groups. Relative liver weight was lower in CIO group. Hepatic triglyceride and cholesterol did not differ among. groups. Fecal excretion of neutral steroid and bile acid were not different among groups, but fecal triglyceride excretion was significantly increased in FO and CI groups compared with the Control group. In conclusion, supplementation of oligosaccharides and chicory inulin at 6% of diets showed no significant hypolipidemic effect in rats fed a high cholesterol diet.

Difference of Classification, Growth and Herbicidal Tolerance in Collected Weedy Rice(Oryza sativa) (수집(蒐集) 잡초성(雜草性)벼(Oryza sativa)의 분류(分類), 생장(生長) 및 제초제(除草劑) 내성차이(耐性差異))

  • Kuk, Y.I.;Guh, J.O.;Chon, S.U.
    • Korean Journal of Weed Science
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    • v.17 no.1
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    • pp.31-43
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    • 1997
  • This study was carried out to investigate classfication of weedy rice (Oryza sativa) based on isozymes esterase and peroxidase, growth and developmental difference of weedy rices and rices grown under dry and water condition, and weedy rice control and tolerant difference of weedy rices in various herbicides using weedy rices collected from thirteen strains of Chonnam, one Chonbuk, two Kyeongki and two rice cultivars. 1. The collected weedy rices were classified into three groups based on isozyme esterase and peroxidase using polyacrylamide gel electrophoresis(PAGE) method. The classified groups were not same each other. 2. Plant height was taller in collected weedy rices than rice cultivars at 18 days after seeding under dry and water conditions, but number of leaves, shoot fresh weight, root fresh weight and root length were not significantly different between collected weedy rices and rice cultivars. In addition, growths of collected weedy rices were greater in dry- than water-condition. 3. After thiobencarb(S-4-chlorobenzyl diethythiocarbamate), molinate(S-ethyl hexahydro-1H-azepine-1-carbothioate) and oxadiazon(5-tert-butyl-3(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one) were applied at 6 days before seeding, the weedy rices controlled 100% by thiobencarb at 2.1kg ai/ha and 024kg ai/ha oxadiazon treatment but controlled 26% to 67% by molinate at 6.5kg ai/ha. Rice due to the herbicides was injured severely(25% to 100%) in flood condition at time of rice seeding after oxadiazon at 0.48kg ai/ha and 2.1kg ai/ha thiobencarb application, except for molinate which injured rice slightly(4% to 13%) in drain condition. The collected weedy rices to all experimented herbicides showed slight intraspecific variations. The intraspecific variations of weedy rices decreased in the order of thiobencarb>molinate>oxadiazon.

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Effect of Exogenous Hormones on Anthocyanin Accumulation and Phenylalaine Ammonia-lyase and Chalcone-synthase Activity in the Hairy Root Cultures of Raphanus sativus cv. Chungpihongsim (청피홍심무우의 모상근 배양에서 안토시아닌의 생성 및 PAL, CHS의 활성에 미치는 호르몬의 영향)

  • 안준철;백윤웅조혜선황백
    • KSBB Journal
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    • v.9 no.1
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    • pp.26-34
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    • 1994
  • When effects of exogenous hormone on hairy root cultures of Raphanus sativs cv. Chungpihongsim examined, the highest anthocyanin synthesis and disorganization were observed when 2, 4-D was supplemented to the culture medium Cytokinins showed early weak induction after transfer and ABA showed inhibitory effect and GA3 showed no effects in anthocyanin synthesis. Hormones except for 2, 4-D in 1 mg/$\ell$ concentration did not induce disorganization of hairy root and retarded growth of hairy root. Time-course changes in anthocyanin synthesis, phenylalanine ammonia-lyase activity and chalcone synthase activity were examined in culture condition contalning 2, 4-D and kinetin. In a medium containing 2, 4-D, anthocyanin synthesis began to increase on the 9th day and reaching maxima on the 18th day after transfer. Maximum peak of PAL activity appeared on the 3-9th day and another minor peak appeared on the 18th day. CHS activity increased from 9th day, reaching maximum on the 18th day and remained at a relatively high level for culture period. In a medium containing kinetin, anthocyanin synthesis increased temporarily on the 6-9th days, early days after transfer and maintained at a low level for remaining culture period. Peak of PAL activity appeared on the 6th day and CHS activity increased from the 6th days, reaching maxima about 18th day and remained at a relatively high level. In particular, addition of kinetin after preculture in hormone free medium for 2 weeks which was thought of wound healing period showed no effects in anthocyanin synthesis. This results showed that stimulation of anthocyanin synthesis by 2, 4-D and kinetin was meaningfully connected with changes of PAL, CHS activity, and then suggested rate-limiting role of CHS on anthocyanin synthesis in that there is close correlation between anthocyanin synthesis and changes of CHS activity in time-course. Besides, it is considered that cytoklnins involving kinetin stimulated anthocyanin synthesis be due to "wound response" by cutting of young roots, and that difference in time-course peak and PAL, CHS activities expressed by 2, 4-D and kinetin result from occurrence of isozyme which have different regulatory mechanism.mechanism.

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Biochemical Characterization of Phospholipase C$\delta$from liver of Mud loach (Misgurnus mizolepis) (미꾸라지 간으로부터 포스포리파아제 C델타 단백질의 생화학적 특성)

  • Seo, Jung-Soo;Lim, Sang-Uk;Kim, Na-Young;Lee, Sang-Hwan;Oh, Hyun-Suk;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.67-80
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    • 2005
  • Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.

Studies on the Resistance of Rice Varieties to Biotypes of the Brown Planthopper, Nilaparvata lugens $ST{\AA}L$ (벼멸구생태형(生態型)에 대한 수도품종(水稻品種)의 저저성(抵抵性)에 관한 연구(硏究))

  • Kim, Jeong-Wha;Kim, Doo-Ho
    • Korean journal of applied entomology
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    • v.24 no.4 s.65
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    • pp.209-218
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    • 1986
  • This study was performed to evaluate the differences in resistance of rice varieties to biotypes of the brown planthopper (BPH), capable of surving on the Milyang 30 and Milyang 63 varieties which have Bph 1 and bph 2 gene for resistance, respectively. The rice varieties tested were Milyang 30, Cheongcheong, Milyang 63 and Gaya which have been reported as having resistant genes for the BPH. The check varieties were Chucheong and Sangpoong which have no resistant gene. The degree of resistance to the BPH biotypes indicated that Milyang 30, Cheongcheong, Milyang 63 and Gaya varieties were highly resistant to the biotype 1. But their reactions against biotype 2 and 3 were variable, namely Milyang 30 and Cheongcheong were susceptible to biotype 2, and Milyang 63 was susceptible to biotype 3. In the esterase isozyme patterns of brown rice the bands of ${\beta}-1,\;{\beta}-3\;and\;{\beta}-5$ were detected in Chucheong and Sangpoong, while the bands of ${\alpha}-1,\;{\beta}-2\;and\;{\beta}-5$ were detected in the test varieties which have genes for resistance. However, the bands of ${\beta}-5$ in Milyang 63 and Gaya were stronger than those of Milyang 30 and Cheongcheong varieties. In the root of 10-days seedling, the esterase bands of ${\alpha}-2,\;{\beta}-2\;{\beta}-4\;and\;{\beta}-5$ were detected in Chucheong and Sangpoong, while the bands of ${\alpha}-1,\;{\beta}-1\;{\beta}-3\;and\;{\beta}-5$ were detected in the tested different varieties. But the bands of ${\alpha}-1\;and\;{\beta}-5$ in Milyang 63 and Gaya were stronger than those of Milyang 30 and Cheongcheong varieties.

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Studies on the Mechanism of Varietal Resistance of Rice to the Brown Planthopper(Nilaparvata lugens $ST{\AA}L$ (벼멸구에 대한 수도품종(水稻品種)의 저항성기작(抵抗性機作)에 관(關)한 연구(硏究))

  • Kim, J.W.;Choi, S.Y.;Park, J.S.
    • Korean journal of applied entomology
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    • v.24 no.2 s.63
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    • pp.51-60
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    • 1985
  • This paper was performed to study the nature of varietal resistance of some Korean-new rice cultivars to the brown planthopper (BPH), Nilaparvata lugens ($ST{\AA}L$). The rice cultivars tested were Cheongcheong, Gaya, Hangangchal, Samgang, Nampoong and Yeongpoong which have been reported as having resistant genes for the BPH. The check varieties were Jinheung, Sangpoong and Chucheong for susceptible and IR-36 for resistant. The factors studied were referred to the seedling responses, preference in feeding and oviposition of BPH, antibiosis (nymphal development, adult emergence and sex ratio, adult body weight, population build-up, feeding amount and amylase activity), and chemical composition (inorganic components, chlorophyll contents, cell wall components, amino acids and esterase isozymes) of leaf- sheath and/or roots of rice plants. In conclusion, the natures of varietal resistance of rice cultivars to the BPH were not only correlated with the resistant reaction of rice plant, but also they were related with the non preference in feeding and oviposition and those resistant cultivars had the antibiotic effects to the insects. Their antibiotic effects of rice cultivars to the BPH would be related with some of the chemical components of rice plants, such as the contents of magnesium oxide (MgO), and chlorophyll and the different esterase isozymes.

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