• Journal of Applied Biological Chemistry
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• 제43권4호
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• pp.240-245
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• 2000

Two genes encoding maltooligosyltrehalose synthase (SaMTS) and maltooligosyltrehalose trehalohydrolase (SaMTH) were isolated from a hyperthermophilic microorganism, Sulfolobus acidocaldarius (ATCC 49462). ORFs of the SaMTS and SaMTH genes are 2,163 and 1,671 bp long and encode 720 and 556 amino acid residues, respectively. A bifunctional fusion enzyme (SaMTSH) was constructed through the gene fusion of SaMTS and SaMTH. Recombinant SaMTS, SaMTH, and SaMTSH fusion enzyme were overexpressed in E. coli BL21. SaMTS and SaMTH produced trehalose and maltotriose from maltopentaose in a sequential reaction. SaMTSH fusion enzyme catalyzed the sequential reaction in which the formation of maltotriosyltrehalose was followed by hydrolysis leading to the synthesis of trehalose and maltotriose. The SaMTSH fusion enzyme showed the highest activity at pH 5.0-5.5 and $70-75^{\circ}C$. SaMTS, SaMTH, and SaMTSH fusion enzyme were active in soluble starch, which resulted in the production of trehalose.

• Journal of Ginseng Research
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• 제24권2호
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• pp.83-88
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• 2000

6년생 고려인삼근(Panax ginseng C.A. Meyer) 중의 Sucrose synthase, UDP-glucose pyrophosphorylase 및 ADP-glucose pyrophosphorylase의 활성을 생육 시기별로 조사한 결과, Sucrose synthase 와 ADP-glucose pyrophosphorylase는 뿌리저장활성 지표로서 adaptive enzyme의 특성을 나타내는 반면, UDP-glucose pyrophosphorylase는 maintenance enzyme으로서 존재하였다. 평균기온이 24。C 이상일 때 전분합성이 저하되고 중심부의 산소소비량이 급격히 증가되었다.

• 한국미생물·생명공학회지
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• 제26권5호
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• pp.442-449
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• 1998

CGTase를 이용한 당전이 xylitol의 합성과 당전이 xylitol의 비피더스균 생육증식 효과에 대한 연구를 수행하였다. 수종의 세균류가 분비하는 CGTase의 xylitol에 대한 당전이능을 비교하였으며, Thermoanaerobacter sp. 유래의 CGTase가 가장 우수한 당전이능을 보였다. 각종 당공여체를 검토한 결과 압출전분이 가장 우수한 결과를 보였으며, 당전이 효소반응의 최적 조건을 검토하였다. 생성된 당전이 xylitol을 활성탄-셀라이트 칼럼 크로마토그래피를 이용하여 분리하여 두 개의 fraction인 F-I, F-II를 얻었다. 이들의 당쇄결합 양상을 FAB mass spectrometer와 $^{13}$C-NMR spectrometer, 그리고 glucoamylase을 이용한 효소소화법을 이용하여 분석한 결과 xylitol에 glucose와 maltose 분자가 $\alpha$-1,4 결합되어 있는 것으로 유추되었다. 얻어진 당전이 xylitol은 xylitol과는 달리 Bifidobacterium breve에 대한 생육촉진효과를 보였다.

• 한국작물학회지
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• 제41권5호
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• pp.569-577
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• 1996

본 실험은 콩(황금콩, 방사콩), 땅콩(남대땅콩, 진풍땅콩) 및 옥수수(진주옥, 수원19호)를 공시재료로 하여 PEG처리에 의한 수분 포텐셜이 발아 및 묘의 생육과 저장양분의 함량변화에 미치는 영향을 구명하기 위하여 실시되었다. 이 실험에서 발아 배지로 사용된 PEG(M.W. 10,000)용액의 수분 포텐셜은 각각 0.0, -0.2, -0.SMPa이었다. 묘의 수분함량, 묘장이 조사되었으며, 단백질, 지방, 전분은 종자의 함량에 대한 감소율과 가용성 당은 증가율로 조사되었다. 1. 세 작물중 콩의 수분흡수율이 가장 높았다. 2. 세 작물 모두 -0.5MPa의 수분포텐셜에서 수분함량이 현저하게 감소하였고 묘의 신장은 수분포텐셜이 감소함에 따라 지연되었다. 3. 수분포텐셜이 감소함에 따라 종자발아에 의한 단백질의 감소율은 콩과 땅콩에서 낮았고, 전분의 감소율은 옥수수의 -0.5MPa 처리에서 낮았다. 4. 수분포텐셜에 의한 당의 증가율은 세 작물 모두 -0.5MPa에서 현저하게 감소하였다. 5. 세 작물 모두에서 발아를 위한 수분의 이용은 -0.5MPa 이하의 수분포텐셜에서 영향을 받으며, 당의 합성에 큰 영향을 미치는 것으로 나타났다

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.227-227
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• 2017

This study was conducted to analyze the reduction of quality and ripening rate of rice due to submergence during ripening stage and identify the physiological cause. Korean japonica type rice cultivars, Nampyeong was used in the experiment. The following 7, 14 day after heading, they were then moved into submergence treatment facility and we conducted the tests under three different submergence conditions - T1(clear water and overhead submergence), T2(Muddy water and exposure of Flag leaf end ), T3(Muddy water and overhead submergence), and the treatment lasted for 4days. The decrease of ripening rate was most severe in T3 treatment at 7days after heading, and the decrease of head rice ratio was most severe in T3 treatment at 14days after heading. Meanwhile the starch synthesis was inhibited, as the supply of assimilation products was inhibited in grain during the submergence treatment. And in stem, sucrose content was increased because the soluble carbohydrates accumulated before heading were decomposed. These changes may be due to the consumption of respiratory substrate in anaerobic conditions and the inhibition of the production of photosynthetic products by light interception. In order to see what kind of reaction occurs at the molecular level, we examined the degree of RNA expression in grain, stem and leaf. First, the expression of rna associated with starch synthesis in grain was decreased in all submergence treatment. and especially Ospul(DBE) was more decreased in 14days after heading treatment than 7days after heading treatment. This difference can explain the result that the decrease of head rice ratio was more severe at 14days after heading. And in stem, the expression of rna associated with the supply of assimilation products was decreased in submergence treatment. Finally in leaf, the expression of rna(ADH, ALDH) associated with anaerobic respiration was increased, while the expression of rna associated with photosynthesis was decreased. These results of physiological analysis can used to develop the cultivation technique and to offer the information for breeding the cultivars with tolerant characteristics to submergence stress during ripening stage in rice.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.59-59
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• 2017

Rice (Oryza sativa L.) is the most important crop and its yield must be improved to feed the increasing global population. Recently developed high-yielding varieties with extra-large sink capacity often have a problem in unstable grain-filling. Therefore, understanding limiting factors for improving grain-filling and controlling them are essential for further improvement of rice grain yield. However, since grain-filling rate was determined by complex sink-source balance, the ability of grain-filling was very difficult to evaluate. Source ability for 'grain' was not only determined by the ability of carbon assimilation in leaves, but also that of carbon translocation from leaves to panicles. Sink strength was determined by the complex carbon metabolism from sucrose degradation to starch synthesis. Hence, to evaluate the grain-filling ability and determine its regulatory steps, the whole picture of carbon flow from photosynthesis at leaves to starch synthesis at grains must be revealed in a metabolite level. In this study, the yield and grain growth rate of three high-yielding varieties, which show high sink capacity commonly, were compared. Momiroman showed lower grain filling rate and slower grain growth rate than the other varieties, Hokuriku 193 and Tequing. To clarify the limiting point in the carbon flow of Momiroman, $CO_2$ labeled by stable isotope ($^{13}C$) was fed to three varieties during ripening period. The ratio of $^{13}C$ left in the stem was higher in Momiroman 24 hours after feeding, suggesting inefficient carbon translocation of Momiroman. More interestingly, $^{13}C$ translocation from soluble fraction to insoluble one in the grain seemed to be slower in Momiroman. To get the further insight in a metabolite level, we are now trying the $^{13}C$ labeling metabolome analysis in the developing grains.

• 한국근적외분광분석학회:학술대회논문집
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• 한국근적외분광분석학회 2001년도 NIR-2001
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• pp.1158-1158
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• 2001

Fast and dynamic biochemical, enzymatic and morphological changes occur during the so-called generative development and during the vegetative processes in seeds. The most characteristic biochemical and compositional changes of this period are the formation and decline of storage components or their precursors, the change of their degree in polymerization and an extensive change in water content. The aim of the present study was to detect the maturation processes in seed nondestructively and to verify the applicability of near infrared spectroscopic methods in the measurement of physiological, chemical and biochemical changes in wheat seed. The amount and variation of different water “species” has been changed intensively during maturation. Characteristic changes of three water absorption bands (1920, 1420 and 1150 nm) during maturation were analysed. It was concluded that the free/bound transition of water molecules could be followed sensitively in different region of NIR spectra. Kinetic changes of carbohydrate reserves were characteristic during maturation. An intensive formation and decline of carbohydrate reserves were observed during early stage of maturation (0 -13 days, high energy demand). An accelerated formation of storage carbohydrates (starch) was detected in the second phase of maturation. Five characteristic absorption bands were analysed which were sensitive indicators the changes of carbohydrates occurred during maturation. Precursors of protein synthesis and the synthesis of reserve proteins and their kinetic changes during maturation were followed from NIR spectra qualitative and qualitatively. Dynamic formation of amino acids and the changes of N forms were detected by spectroscopic, chromatographic and by capillary electrophoresis methods. Calibration equations were developed and validated in order to measure the optimal maturation time protein and moisture content of developing wheat seeds. The spectroscopic methods are offering chance and measurement potential in order to detect fine details of physiological processes. The spectra have many hidden details, which can help to understand the biochemical background of processes.

• Journal of Microbiology and Biotechnology
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• 제33권10호
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• pp.1337-1350
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• 2023

Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35℃, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.

• Journal of Microbiology and Biotechnology
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• 제18권9호
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• pp.1544-1549
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• 2008

MhMTS and MhMTH are trehalose ($\alpha$-D-glucopyranosyl-[1,1]-$\alpha$-D-glucopyranose) biosynthesis genes of the thermophilic microorganism Metallosphaera hakonensis, and encode a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively. In this study, the two genes were fused in-frame in a recombinant DNA, and expressed in Escherichia coli to produce a bifunctional fusion enzyme, MhMTSH. Similar to the two-step reactions with MhMTS and MhMTH, the fusion enzyme catalyzed the sequential reactions on maltopentaose, maltotriosyltrehalose formation, and following hydrolysis, producing trehalose and maltotriose. Optimum conditions for the fusion enzyme-catalyzed trehalose synthesis were around $70^{\circ}C$ and pH 5.0-6.0. The MhMTSH fusion enzyme exhibited a high degree of thermostability, retaining 80% of the activity when pre-incubated at $70^{\circ}C$ for 48 h. The stability was gradually abolished by incubating the fusion enzyme at above $80^{\circ}C$. The MhMTSH fusion enzyme was active on various sizes of maltooligosaccharides, extending its substrate specificity to soluble starch, the most abundant natural source of trehalose production.

• Applied Biological Chemistry
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• 제41권4호
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• pp.222-227
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• 1998

The activities of enzymes of carbohydrate metabolism have been determinated in the host cytosolic and bacteroid fractions of cowpea (Vigna unguiculata) nodules formed with B. japonicum I 16 and in roots of nodulated cowpeas. The host cytosolic fraction of the nodules contained the enzymes of glycolytic pathway and the pentose phosphate pathway, whereas the bacteroids had only limited capacity for carbohydrate metabolism and appeared to be insufficient for the complete glycolytic pathway as well as starch synthesis and degradation. In a time-course study, using plants grown in a glasshouse, the acetylene-reducing activity (ARA) of the nodules increased in parallel with the total N content of plants and protein of the nodules until approximately 8 weeks after planting. Subsequently, the weight and size of the nodules and the weight of the plants continued to increase, but there was a sharp decrease in the ARA and the total N content of the plants.