• Applied Biological Chemistry
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• 제11권
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• pp.43-47
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• 1969

본(本) 연구(硏究)에서 얻어진 결과(結果)를 요약(要約)하면 다음과 같다. 1. Glucose Isomerizing Enzyme을 분비(分泌)하는 Actinomyces과(科) 균주(菌株)를 토양(土壤)으로부터 분리(分離)하였으며 2. 본(本) 균주(菌株)가 생성(生成)하는 Glucose Isomerizing Enzyme를 Glucose에 작용(作用)시킨 결과(結果) 생성(生成)된 당(糖)은 Fructose 일종(一種)만을 생성(生成)하였으며 그외(外)의 당(糖)의 부생물(副生物)은 검출(檢出)되지 아니 하였다.

• 한국미생물·생명공학회지
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• 제20권4호
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• pp.437-444
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• 1992

분쇄마찰매체함유 불균일상 효소반응계를 활용한 생전분의 무증자당화법은 고농도, 고순도의 포도당을 고효율로 직접 얻을 수 있는 새로운 당화법으로 이를 high fructose corn syrup(HFCS) 제조공정에 응용코져 연구하였다. 400g/$\ell$ 생전분을 24시간 무증자당화시켜 이성화반응에 적합한 고농도인 398g/$\ell$와 고순조인 98 포도당 용액을 농축과정을 거치지 않고 직접 얻을 수 있었다. 전분에 대한 포도당수율은 0.90으로, 미반응 잔유전분은 쉽게 원심분리되었으며, 재당화도 용이하였다. 또한 무증자당화액은 단백질성 변성물, 이온 등 불순물의 함량이 매우 적어, 이성화 반응 전단계인 분리 정제과정을 단순화할 수 있었다. 생성된 당용액의 고정화 포도당 이성화효소 기질로서의 적합성을 검토하여 우수한 결과를 얻었다. 생전분 직접당화법을 도입한 새로운 HFCS 제조공정은 액화/당화공정을 경유하는 기존의 공정에 비하여 여러가지 장점이 있어 산업적 활용이 기대되며, 이를 위한 후속연구가 요망된다.

• Molecules and Cells
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• 제28권4호
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• pp.397-401
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• 2009

Flavonoids are a group of polyphenolic compounds that have been recognized as important due to their physiological and pharmacological roles and their health benefits. Glycosylation of flavonoids has a wide range of effects on flavonoid solubility, stability, and bioavailability. We previously generated the E. coli BL21 (DE3) ${\Delta}pgi$ host by deleting the glucose-phosphate isomerase (Pgi) gene in E. coli BL21 (DE3). This host was further engineered for whole-cell biotransformation by integration of galU from E. coli K12, and expression of calS8 (UDP-glucose dehydrogenase) and calS9 (UDP-glucuronic acid decarboxylase) from Micromonospora echinospora spp. calichensis and arGt-4 (7-O-glycosyltransferase) from Arabidopsis thaliana to form E. coli (US89Gt-4), which is expected to produce glycosylated flavonoids. To test the designed system, the engineered host was fed with naringenin as a substrate, and naringenin 7-O-xyloside, a glycosylated naringenin product, was detected. Product was verified by HPLC-LC/MS and ESI-MS/MS analyses. The reconstructed host can be applied for the production of various classes of glycosylated flavonoids.

• KSBB Journal
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• 제19권5호
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• pp.406-409
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• 2004

Matrix Assisted Laser Desorption ionization Time of Flight (MALDI-TOF) was used for enzymes identification related to B -1,3-glucan synthesis. Agrobacterium sp. ATCC31750 was cultivated with two stage Continuous Stirrer Tank Reactor (CSTR) and the cells were harvested and their protein profiles were analysed by two dimensional electrophoresis. The specific enzyme spot was treated with trypsin and ana lysed by MALDI-TOF to get peptide molecular weight. The peptide molecular weights were matched with Agrobacterium tumefacience's Data Base from the matrix science site, then could identify the avaliable key enzymes. In this study, we identified key metabolite of synthesis of beta-1,3-glucan, such as glucose-6-phosphate isomerase, phosphoglucomutase, B-1,3-glucan synthase and glucokinase, and we also identified uracil phosphoribocyl transferase and Ribosome recycling factor also.

• 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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• 제55권2호
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• pp.121-128
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• 2017

Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected ($V^+$) and uninfected ($V^-$) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in $V^+$ compared with $V^-$ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in $V^+$ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in $V^+$ and $V^-$ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.

• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.606-612
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• 2018

The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

• Applied Biological Chemistry
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• 제39권6호
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• pp.443-448
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• 1996

xylA 유전자의 프로모터상에서 조절양상을 조사하기 위하여 xylA 유전자의 프로모터(Pxyl)와 lacZ 유전자를 연결한 Pxyl-lacZ 융합 유전자를 제작하여 xylose에 의한 ${\beta}-galactosidase$ 생산의 조절양식을 조사하였다. xylA 프로모터 부위를 분리하여 lac 프로모터가 없는 고복제수의 lac 오페론 백터인 pMC1403에 클로닝시켜 pMCX191을 제작하여 reading frame에 변화가 없는 Pxyl-lacZ 융합 유전자를 만들었으며 이 벡터에서 Pxyl-lacZ 단편을 분리한 후 저복제수 벡터인 pLG339에 클로닝시켜 pLGX191을 제작하였다. 상기 플라스미드들을 xylA 변이주인 DH77에 형질전환시켜 Pxyl-lacZ 융합 유전자에서 ${\beta}-galactosidase$의 발현조절을 조사한 결과 xylose 농도에 따른 유도, glucose에 의한 발현억제 및 cAMP에 의한 억제해제 양상 등이 염색체상의xylA 유전자의 발현조절과 같은 경향을 나타내었다. pMCX191과 pLGX191을 이용하여 유전자 투여 량 효과를 본 결과도 복제수에 따른 차이가 크지 않았다. xylA 프로모터 부위내 조절영역를 추정하기 위해 구조유전자 상류 -209 bp를 포함한 xylA 유전자를 pUC19에 클로닝시킨 pUX30에서 프로모터 부위가 부분결손된 벡터들을 제작하여 결손부위의 염기서열을 확인하였다. 이들 부분결손 xylA 프로모터를 가진 xylA 유전자에서 xylose isomerase의 발현을 조사한 결과, 번역 개시점에서 -166 bp 이상의 영역을 결손시킨 pUX31과 pUX32의 경우pUX30과 비슷한 발현 양상을 보인 반면 -120 bp 이상의 영역을 결손시킨 pUX33과 pUX34에서는 모벡터에 비해 약 30% 수준의 발현을 보였다. 또한 pUX33과 pUX34에서는 xylose 유도시 cAMP에 의한 발현 촉진효과도 볼 수 없었다. -59 bp 이상의 부위가 결손된 pUX35의 경우에는 전혀 xylA 유전자의 발현이 일어나지 않았다. 이러한 결과로 볼때 xylA 프로모터내의 조절부위는 -165 bp에서 -59 bp 사이에 존재하는 것으로 추정되었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제5권2호
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• pp.205-211
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• 2002

Previously, genetic relationships among 321 silkworm races preserved in the Department of Sericulture and Entomology, NIAST, RDA were studied using six isozymes comprising 20 loci. As a part of additional studies, three additional isozymes with 7 loci [phos-phoglucomutase (PGM), glucose phosphate isomerase (GPI), malete dehydrogenase (MDH)] from hemolymphs, midgets, and eggs were employed to reevaluate the previous dendrogram (UPGMA method). Although the current study supported the recognition of the previously identified major groups, some minor changes were apparent among the selected 47 forms. The current study showed that the polymorphic loci of GPI from eggs and MDH from hemolymph appear to be responsible characters for generating a new major group in the genetic relationship. Interpreting the data with current additive isozymes may represent more robust genetic relatioships among 321 silkworm races preserved in Korea, until further evidence is available.

• 한국패류학회지
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• 제11권1호
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• pp.51-61
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• 1995

A horizontal starch gal electrophoresis for enzyme proteins extracted from 3 species of Korean planorbid snails; Gyraulus convexiusculus, Hippeutis cantori and Segmentina hemisphaerula was carried out in order to elucidate their genetic relationships.The results from 12 enzymes employed in three different kinds of buffer systems are summarized as follows:1) Two loci from each enzyme of aldehyde oxidase, esterase, glucose phosphate isomerase. isocitrate dehydrogenase, leucine aminopeptidase, malate dehyogenase, peptidase and xanthine oxidase were detected, and only one locus was observed from each of the following four enzymes: 6-phosphogluconate dehydrogenase, glyceraldehyde-phosphate dehydrogenase, glutamate oxaloacetate transaminase and glycerol-3-phosphate dehydrogenase.2) Most of loci in 3 species of planorbid snails employed showed homozygous and monomorphic banding patterns and some of them were specifis as genetic markers among different species. However, a few of loci (EST-1. EST-2 and GPI-2)showed polymorphic banding patterns. 3)Hippeutis cantori and Segmentina hemisphaerula were more closely clustered in a dendrogram with the genetic iddentity value of 0.431, and these two species were lineated with Gyraulus convexiusculus as another cluster at the value of 0.294.In summarizing the above results, three species of Korean planorbid snails employed in this study mostly showed monomorphic enzyme protein banding patterns and genetic differences specific among 3 species.