• 한국가축번식학회지
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• 제20권3호
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• pp.251-258
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• 1996

본 연구는 착성전 Rat 8-세포기 난자의 체외배양시 glucose95.56 mM와 0mM)와 phosphate(1.19mM와 0 mM)의 억제효과를 규명하기 위하여 실시하였다. 48시간 배양후 발육율은 glucose＋phosphate군에서는 37%(31/84), glucose군은 70%(64/91), phosphate군은 69% (59/85) 그리고 glucose와 phosphate가 없는 무처리군에서는 77%(67/85)를 나타냈다. Glucose＋phosphate군의 발육율은 다른 처리군에 비해 유의적으로 낮았고, 그 외 처리군 사이에는 유의성이 없었다. 발육난자의 핵염색에 의한 세포수의 측정에서는 glucose군에서 가장 많은 세포수를 나타냈고 (29.3$\pm$0.97, P<.001), 그외 처리에서는 유의성이 없었다 (glucose＋phosphate, 17.5$\pm$1.04; phosphate, 18.6$\pm$1.01; no glucosephosphate, 19.8$\pm$1.01). 본 실험의 결과로 glucose와 phosphate는 Rat 8-세포기 난자의 발육에 억제효과를 나타냈고, glucose는 난자의 세포분열을 증진시키는 효과가 있는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.685-690
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• 2007

The deoxysugar biosynthetic gene cluster of Sch 47554/Sch 47555 was cloned from Streptomyces sp. SCC-2136. One of the ORFs, schS6, appeared to encode glucose-1-phosphate thymidylyltransferase, which converts dTTP and glucose-1-phosphate to TDP-D-glucose and pyrophosphate. The dTDP-D-glucose is a key metabolite in prokaryotics as a precursor for a large number of modified deoxysugars, and these deoxysugars are a maj or part of various antibiotics, ranging from glycosides to macrolides. SchS6 was expressed in E. coli vector pSCHS6 and the expressed protein was purified to apparent homogeneity by ammonium sulfate precipitation and Ni-NTA affinity column chromatography. The specific activity of the purified enzyme increased 4.7-fold with 17.5% recovery. It migrated as a single band on SDS-PAGE with an apparent molecular mass of 56kDa. The purified protein showed glucose-1-phosphate thymidylyltransferase activity, catalyzing a reversible bimolecular group transfer reaction. In the forward reaction, the highest activity was obtained with combination of dTTP and ${\alpha}-D-glucose-1-phosphate$, and only 12% of that activity was obtained with the substrates $UTP/{\alpha}-D-glucose-1-phosphate$. In the opposite direction, the purified protein was highly specific for dTDP-D-glucose and pyrophosphate.

• 농업과학연구
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• 제22권1호
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• pp.96-102
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• 1995

본 연구는 돼지 체외수정란의 발육에 있어서 적절한 에너지 급원을 구명하기 위하여 수행하였는 바, BMOC-II 배양액을 기본배양액으로 하여 여기에 해당과정을 통하여 에너지를 생산하는 glucose와 이 과정에 중요하게 작용하는 phosphate를 실험설계에 따라 첨가한 배 양액에 체외에서 발생된 돼지의 수정란을 배양하여 수정란의 발육에 미치는 영향을 조사하였다. 1. 배양액에 Glucose와 phosphate가 모두 첨가되지 않은 배양액에서 배양된 돼지의 수정란은 15.2%가 4-세포란 이상으로 발육하였으나, 상실배로는 전혀 발육하지 못하였다. 2. 배양액에 첨가된 glucose의 농도는 phosphate의 첨가유무에 관계없이 농도가 높은 처리에서 4-세포기 이상으로 발달한 비율은 감소하였으나, 상실배에 도달한 비율은 증가하는 경향을 나타냈고, phosphate와 함께 첨가되었을 때보다 단독으로 첨가하였을 때가 첨가농도에 관계없이 돼지 체외수정란의 발육능을 증가시켰다.

• BMB Reports
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• 제34권3호
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• pp.230-236
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• 2001

dTDP-rhamnose is synthesized from dTTP and glucose-1-phosphate by four enzymatic steps in the gram-negative bacteria. By using a homologous PCR product, a gene cluster encoding four genes (rfbA, rfbB, rfbC, rfbD) involved in L-rhamnose biosynthesis by Acinetobacter calcoaceticus was isolated and sequenced. The four genes were clustered on the biosynthetic operon in the order of rfbB, D, A, C. A gene, rfbA, encoding glucose-l-phosphate thymidylyltransferase (RfbA), was cloned from A. calcoaceticus pathogenic and encapsulated in the gram-negative bacterium. This enzyme catalyzes the formation of dTDP-D-glucose From $\alpha$-D-glucose-1-phosphate and dTTP.RfbA was amplified by PCR and inserted into the $T_7$ expression system. The activity of RfbA was determined by the capillary electrophoresis. The $K_m$ values for dTTP and $\alpha$-D-glucose-1-phosphate were calculated to be 1.27 mM and 0.80 mM, respectively by using the Line-Weaver Burk plot. RfbA is inactivated by diethylpyrocarbonate.

• KSBB Journal
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• 제22권5호
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• pp.345-350
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• 2007

인산당은 모든 유기체에서 발견되며 무척 다양한 유용성을 지니고 있다. 특히 glucose 1-phosphate (G1P), glucose 6-phosphate (G6P), fructose 6-phosphate (F6P) 등은 해당과 정, 당합성과정, 5탄당 인산화과정 및 캘빈회로와 같은 탄수화물 대사와 에너지 생산 대사의 주요한 핵심 중간물질이다. 특히 해당과정에서 F6P는 G6P의 이성질화반응에 의하여 생성된다. F6P의 대량생산은 전분을 이용하는 것이 가능한데, 우선 전분에 인산화효소를 가하여 G1P를 얻고, 이 G1P를 자리옮김효소 (phosphoglucomutase, GM)와 이성질화효소 (phosphoglucoisomerase, GI)를 순차적으로 적용하여 G6P와 F6P를 생산하게 된다. 효소반응의 경우 전분의 용해도 증가, 반응속도의 향상 및 미생물의 오염방지 등을 위하여 중온성 효소보다는 고온성 효소 혹은 내열성 효소가 선호된다. 본 연구는 세 가지 내열성 효소를 이용하여 전분으로부터 두 단계반응으로 F6P를 생산하는 것에 관한 것이다. 실험에 사용된 효소는 대장균에서 발현된 재조합 효소로서, 효소의 생산은 유가식 배양을 이용하였다. 1.2% 가용성 전분 200 L를 이용하여 1,253 g의 순수한 G1P를 생산하였으며 이를 이용하여 최종적으로 30% 수율로 F6P를 생산할 수 있었다. 최대수율을 얻기 위하여 반응표면분석법을 이용하여 GM : GI = 1 : 1.23, 63.5$^{\circ}C$, pH 6.85의 조건이 도출되었으며, 이 조건하에서 실험을 통하여 20 g/L의 전분을 이용하여 30% 수율로 F6P가 생성됨을 확인할 수 있었다.

• 한국미생물·생명공학회지
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• 제21권6호
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• pp.570-576
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• 1993

For the production of glucose-1-phosphate from sucrose, bacteria having sucrose phosphorylase were isolated from Kimchi. Among them, JS-05, newly isolated strain having high activity of sucrose phosphorylase was selected and identified as Leuconostoc sp. The specific activity of sucrose phosphorylase of Leuconostoc sp. JS-05 was the highest when the strain was cultured at 25C for 20 hrs in the medium (pH 7.5) containing 10 g sucrose, 5g corn steep liquor, and 2.5g yeast extract per liter.

• 한국동물학회지
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• 제7권2호
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• pp.25-36
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• 1964

The present paper deals with the comparative study on phylogenic difference in the patterns of energy metabolism of brain slices of several vertebrate species by measuring oxygen consumptionwith glucose-6-phosphate, glucose-1-phosphate, glyceraldehyde-3-phosphate or glutamate as respiratory substrate employing Warburg's manometric method, by determination of the utilization rate of glucose using glucose-1-C14 by analyzing patterns of free amino acid distribution , and by histochemical determination using glucose-1-C14 by analyzing patterns of free amino acid distribution acid distribution , and by histochemical determination of glycogen contents. 1. Glucose enhances the oxygen consumption of brain slices of animals belinging to reptile, aves and mammalia while it shows a tendency to decrease that of animals belonging to pisces and amphibia. 2. Glucose-6--phosphate increase oxygen consumption more than glucose in every species examined, while glucose-1-phosphate and glyceraldehyde-3-phosphate increase that of Rana nigromaculata only . In general m, it appears that phosphosugars are more effective as a respiratory substrate to those species which have less endogenous respiration than to those having larger endogenous respiration. 3. Similar patterns of free amino acid distribution and the relative amount are found among the species and in every species examined glutamic acid is detected in the larges amount . ${\gamma}$-Amino butyric acid, glycine, alanine and aspartic acid are found in every species. 4. Ophicephalus showed less oxygen consumption than endogenous respiration when glutamate was added to the medium. When sodium fluoride was added, the oxygen consumption was some what increased . Such phenomenon wasnot found in the frog. 5. The result of histochemcial analysis of the brain showed that glycogen was abundantly present in the fish , amphibia , and especially in the reptile and that no distinctive grains of glycogen were found in the bird and mammal . From these facts, it may be supposed that anaerobic glycolysis as energy source dominates in fish and amphibia and aerobic respiration through the oxidation of glucose dominates in bird and mamal , the reptile occupying transitional position between these two categories. The way of obtaining energy for brain activity by the oxidation of glucose supplied from the circulating blood is seemed to be first acquired by reptile and the function is completed both in aves and mammal.

• Reproductive and Developmental Biology
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• 제28권2호
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• pp.101-105
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• 2004

This study was carried out to evaluate the effects of glucose and sodium phosphate on in vitro development of porcine oocytes matured and fertilized in vitro. When the culture medium was supplemented with various concentrations of glucose, the higher proportions (23 and 26%) of oocytes developed to morular or blastocyst stages were at the concentrations of 2.78 and 5.56 mM than 0 (9%; P<0.05) and 11.12 mM (18%). In experiment to evaluate effect of sodium phosphate during in vitro development of porcine oocytes, a significantly (P<0.05) higher proportions of embryos developed to morular or blastocyst stages was obtained with sodium phosphateof 0.28 (25%) and 0.53 (27％) mM than 0 (15%), 1.05 (19%) and 2.10 (10%) mM. On the other hand, when oocytes were cultured in medium with (0.53 mM) sodium phosphate, the proportions of developed embryos were significantly (P<0.05) higher in medium without (29%) that than with (14%) 5.56 mM glucose. However, a higher proportion of embryos developed to morular or blastocyst stages were obtained in medium with (23%) that than without (8%) glucose (P<0.05). The minimum essential medium (MEM) added to the culture medium were higher regardless of presence of sodium phosphate and glucose on the development of embryos. Although sodium phosphate and glucose could support morular and blastocyst development to a limited extend (10∼24%), significantly higher proportion (36%) at morular or blastocyst stages was obtained by MEM adding in the medium with sodium phosphate and glucose. These results suggest that the early development of in vitro fertilized porcine oocytes can be maintained efficiently by glucose and sodium phosphate when they were cultured in medium with MEM.

• BMB Reports
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• 제37권4호
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• pp.503-506
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• 2004

Glucose-1-phosphate uridylyltransferase from E. coli K12 was used to convert uridine-5'-triphosphate and glucose-1-phosphate to UDP-D-glucose. The conversion was efficient and completed within 5 minutes under the employed conditions. In addition, thymidine-5'-monophosphate kinase and acetate kinase were proven to be non-specific, converting udridine-5'-monophosphate to uridine-5'-triphosphate with 55% conversion after 6 h, which was much slower than the production of TTP under the same conditions (complete conversion within one hour). Since these two reactions could proceed under the same conditions, a one-pot synthesis of UDP-D-glucose with ATP regeneration was designed from easily available starting materials, and conversion up to 40% by HPLC peak integration was achieved given a reaction time of 4 h.

• 미생물학회지
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• 제34권4호
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• pp.207-211
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• 1998

메탄올을 이용하여 성장하는 Methylovorus sp. strain SS1은 formaldehyde의 산화를 위한 linear route의 주효소인 $NAD^+$-linked formaldehyde dehydrogenase 및 $NAD^+$-linked formate dehydrogenas와 cyclic route의 주효소인 hexulose-6-phosphate synthase, glucose-6-phosphate isomerae, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase 등의 활성을 나타내었는데, cyclic route에 관여하는 효소의 활성이 상대적으로 더 높았다. 이 세균은 formaldehyde의 동화와 관련된 ribulose monophosphate 경로의 주효소와 Entner-Doudoroff 경로의 주효소 및 transaldolase 활성과 함께 세포외 다당류 합성과 관련된 phosphoglucomutase, UDP-glucose pyrophyosphorylase, mannose-6-phosphate isomerase의 활성도 나타내었다. 2.3 mM의 ammonium sulfate가 포함된 배지에서 성장한 세균은 7.6 mM의 ammonium sulfate가 포함된 배지에서 성장한 세균보다 더 많은 세포외 다당류를 생산하였지만 균체 수율은 낮았고, 6-phosphogluconate dehydrogenase와 phosphoglucomutase 및 UDP-glucose pyrophoshorylase의 활성은 높게 나타내었으나 6-phosphogluconate dehydratase/2-keto-3-deoxy-6-phosphogluconate aldolase의 활성은 낮았다.