• 한국균학회지
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• 제27권5호
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• pp.328-336
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• 1999

Isozyme comparisons of mycelial extracts from Pleurotus ostreatus were undertaken using isoelectric focusing. Enzyme isozyme patterns were Used to describe the extent of geographical diversity and degree of intraspecific variation in these extracts. A total of 77 bands were resolved from six different enzymes. Cluster analyses were performed using the zymograms for esterase (EST), glucose phosphate isomerase (GPI), leucine aminopeptidase (LAP), malate dehydrogenase(MDH), peroxidase (POX), and phosphoglucomutase (pGM). EST gave multiple banding patterns, while less variability was observed for GPI, MDH, and PGM. Cluster analyses demonstrated that strains of P. ostreatus from geographically different origins are genetically divergent, supporting the idea that there is little or no gene flow between these geographically distant population groups.

• Parasites, Hosts and Diseases
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• 제38권1호
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• pp.45-48
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• 2000

An enzyme analysis of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China was conducted using a horizontal. starch gel electrophoresis in order to elucidate their genetic relationships. A total of eight enzymes was employed from two different kinds of buffer systems. Two loci from each enzyme of aconitase and esterase (${\alpha}-Na{\;}and{\;}{\beta}-Na$) : and only one locus each from six enzymes, gluucose-6-phosphate dehydrogenase (G6PD), ${\alpha}-glycerophosphate$ dehydrogenase (GPD), 3-hydroxybutyrate dehydrogenase (HBDH), malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphoglucomutase (PGM) were detected. Most of loci in two populations of C. sinensis showed homozygous monomorphic banding patterns and one of them, GPD was specific as genetic markers between two different populations. However, esterase (${\alpha}-Na$), GPD, HBDH and PGI loci showed polymorphic banding patterns. Two populations of C. sinensis were more closely clustered within the range of genetic identity value of 0.998-1.0. In summarizing the above results, two populations of C. sinensis employed in this study showed mostly monomorphic enzyme protein banding patterns, and genetic differences specific between two populations.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1200-1210
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• 2004

Metabolic flux analysis was established by adapting previous stoichiometric model developed during growth with cellulose to cell grown with cellobiose for further direct comparison of the bacterial metabolism. In carbon limitation with cellobiose, a shift from acetate-ethanol fermentation to ethanol-lactate fermentation is observed and the pyruvate overflow is much higher than with cellulose. In nitrogen limitation with cellobiose, the cellodextrin and exopolysaccharide overflows are much higher than on cellulose. In carbon and nitrogen saturation with cellobiose, the cellodextrin, exopolysaccharide, and free amino acids overflows reach the highest levels observed but all remain limited on cellulose. By completely shunting the cellulosome, the use of cellobiose allows to reach much higher carbon consumption rates which, in return, highlights the metabolic limitation of C. cellulolyticum. Therefore, the physical nature of the carbon source has a profound impact on the metabolism of C. cellulolyticum and most probably of other cellulolytic bacteria. For cellulolytic bacteria, the use of soluble carbon substrate must carefully be taken into consideration for the interpretation of results. Direct comparison of metabolic flux analysis from cellobiose and cellulose revealed the importance of cellulosome, phosphoglucomutase and pyruvate-ferredoxin oxidoreductase in the distribution of carbon flow in the central metabolism. In the light of these findings, future directions for improvement of cellulose catabolism by this bacterium are discussed.

• Journal of Microbiology and Biotechnology
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• 제9권1호
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• pp.84-90
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• 1999

The growth of Bifidobacterium breve strain HP2 was completely inhibited by the addition of lactate higher than 4.0％ but not by the addition of acetate. Two kinds of lactate-tolerant mutants were isolated by the nitrosoguanidine treatment, enrichment on a liquid medium with 5％ lactate, and selection on agar plates with 5％ lactate. The mutants were not only able to grow in the presence of 5％ lactate but also improved in viable cell stability in the acidic pH range. In a pH-controlled fermentor, mutant N-1-5 grew at a rate slower than that of the wild type but its growth yield was higher. Notably, mutants were more halotolerant and more osmotolerant than the wild type and they were able to grow in the presence of 3％ NaCl or 25％ lactose at which the wild type entirely stopped the growth. The enzyme activities involved in the lactose metabolism in B. breve were measured to elucidate the biochemical basis for lactate tolerance. In the mutants, activities of several enzymes including phosphoglucomutase decreased compared to the wild-type, which may explain their lower growth rate. However, the activity of lactate dehydrogenase or its nature of inhibition by lactate was not altered.

• Parasites, Hosts and Diseases
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• 제50권4호
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• pp.361-364
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• 2012

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.

• 한국패류학회지
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• 제14권1호
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• pp.33-40
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• 1998

A horizontal starch gel electrophoresis for enzyme proteins extracted from 2 species of Korean viviparid snails; Cipangopaludina chinensis malleata and C. japonica was carried out in order to elucidate their genetic relationships. A total of 10 enzymes were employed in three different kinds of buffer systems. Two loci from each enzyme of alcohol dehydrogenase, esterase, glucose phosphate isomerase, isocitrate dehydrogenase, iditol dehydrogenase, malate dehydrogenase and peptidase(VL); and only one locus dach from two enzymes, glycerlo-3-phosphate dehydrogenase and phosphoglucomutase were detected; but, four loci from peptidase(LGG) were observed. Most of loci in two viviparid species showed homozygous monomorphic banding patterns and some of them were specific as genetic markers between two different species. However, EST-1, MDH-1, PEP(VL)-1loci showed polymorphic banding patterns. Foru populations of C. chinensis malleata were more closely clustered in a dendrogram within the range of genetic identity values of 0.928-1.00, and these clusters were lineated with C. japonica at the value of 0.355. In summarizing the above results, two viviparid snail species dmployed in this study mostly showed monomorphic enzyme protein banding patterns, and genetic differences specific between two species.

• Journal of Microbiology and Biotechnology
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• 제29권5호
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• pp.758-764
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• 2019

${\beta}$-Glucan is a chief structural polymer in the cell wall of yeast. ${\beta}$-Glucan has attracted intensive attention because of its wide applications in health protection and cosmetic areas. In the present study, the ${\beta}$-glucan biosynthesis pathway in S. Cerevisiae was engineered to enhance ${\beta}$-glucan accumulation. A newly identified bacterial ${\beta}-1$, 6-glucan synthase GsmA from Mycoplasma agalactiae was expressed, and increased ${\beta}$-glucan content by 43%. In addition, other pathway enzymes were investigated to direct more metabolic flux towards the building of ${\beta}$-glucan chains. We found that overexpression of Pgm2 (phosphoglucomutase) and Rho1 (a GTPase for activating glucan synthesis) significantly increased ${\beta}$-glucan accumulation. After further optimization of culture conditions, the ${\beta}$-glucan content was increased by 53.1%. This study provides a new approach to enhance ${\beta}$-glucan biosynthesis in Saccharomyces cerevisiae.

• Korean Journal of Acupuncture
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• 제29권2호
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• pp.260-270
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• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• 한국수산과학회지
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• 제29권2호
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• pp.256-261
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• 1996

본 연구는 우리나라의 대표적인 연어과 어류인 은연어, 무지개송어 및 그 잡종3배체의 부화자어에 있어서의 동위효소를 분석하여 부화후 초기의 유전적 특징과 유전현상을 규명하고 이를 토대로 이들 종 및 잡종의 식별을 위한 genetic marker를 찾아보고자 5개의 동위효소에 대한 분석을 실시하였다. 이들 중 PGI에서는 종간 차이를 나타내지 않은 반면에 LDH, MDH, IDH 및 PGM 등은 은연어와 무지개송어간에 종 특이적 pattern을 나타내었는데 이는 이들 어종간의 발생 초기에 종의 식별을 위한 유전적 marker로써 유용한 것으로 판단되며, 또한 잡종 및 잡종3배체의 genetic marker로서도 상기 네 가지의 동위효소는 유용한 것으로 나타났다.

• Molecules and Cells
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• 제37권2호
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• pp.172-177
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• 2014

A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-${\alpha}$-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4' and 7 hydroxyl groups, but not at the $5^{th}$ hydroxyl group of the A-ring, resulting in the production of genistein 4'-O-${\beta}$-D-glucoside, genistein 7-O-${\beta}$-D-glucoside (genistin), genistein 4',7-O-${\beta}$-D-diglucoside, biochanin A-7-O-${\beta}$-D-glucoside (sissotrin), daidzein 4'-O-${\beta}$-D-glucoside, daidzein 7-O-${\beta}$-D-glucoside (daidzin), daidzein 4', 7-O-${\beta}$-D-diglucoside, and formononetin 7-O-${\beta}$-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOF-ESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli $BL21(DE3)/{\Delta}pgi{\Delta}zwf{\Delta}ushA$ overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of $200{\mu}M$ of supplemented isoflavonoids, without any additional UDP-${\alpha}$-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides.