• 제목/요약/키워드: Trehalose synthesis

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느타리버섯균의 Trehalose 합성(合成)(I) -배양(培養) 조건(條件)- (Synthesis of Trehalose by Pleurotus spp. -Cultural conditions­-)

  • 홍재식;윤세억;김영수;이종배
    • 한국균학회지
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    • 제15권2호
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    • pp.108-115
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    • 1987
  • 합성배지(合成培地)에서 P. sajor-caju와 P. osreatus 201의 배양조건(培養條件)을 달리하여 배양(培養)할 때 trehalose 합성(合成)과 균체양(菌體量)의 변화(變化)를 조사(調査)한 결과(結果)는 다음과 같다. 1. Trehalose 합성(合成)에 적합(適合)한 탄소원(炭素源)은 양균주(兩菌株) 모두 glucose였고 그 다음이 maltose였으며 glucose 최적농도는 1%였다. 균체수률(菌體收率)이 높은 탄소원(炭素源)은 glucose, maltose 순이었다. 2. Trehalose 합성(合成)에 적합(適合)한 탄소원(炭素源)은 양균주(兩菌株) 모두 peptone 이었고 그 농도(濃度)는 0.05%였으며 균체수률(菌體收率)은 peptone, casamino acid, $(NH_4)_2HPO_4$ 순으로 높았다. 3. Trehalose 합성(合成)에 적합(適合)한 $NH_2HPO_4$$MgSO_47H_2O$의 농도(濃度)는 양균주(兩菌株) 모두 0.1%, 0.04% 였고 균체수률(菌體收率)은 $KH_2PO_4$ 0.2%, $MgSO_47H_2O$ $0.04{\sim}0.08%$가 높았다. 4. Trehalose 합성(合成)에 적합(適合)한 온도(溫度)와 pH는 $25^{\circ}C$, pH5.5였고 균체수률(菌體收率)은 $30^{\circ}C$, pH5.5에서 높았으며 trehalose 합성(合成)은 10일간(日間) 배양(培養)했을 때 균체수률(菌體收率)은 14일간(日間) 배양(培養)했을 때가 가장 양호(良好)하였다.

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배추 trehalose 합성 유전자와 연관된 DNA 마커 개발 (Development of DNA Markers for Trehalose Synthesis Genes in Brassica rapa L.)

  • 정예솔;임용표;허윤강;정상민
    • 생명과학회지
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    • 제19권5호
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    • pp.639-643
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    • 2009
  • 배추는 재배 온도에 따라 수확량과 품질에 많은 영향을 받을 수 있다. 고온 스트레스 저항성 배추품종 육성을 위해 본 연구는 스트레스 저항성에 관련이 있다고 알려진 trehalose 유전자 관련 다형성 마커를 개발하였다. 아라비돕시스 trehalose 합성 유전자와 유사한 총 28개의 배추 EST를 NCBI database에서 찾고 고온에 상대적으로 약한 지부품종과 고온에 잘 견디는 품종인 권심을 대상으로 다형성을 조사하였다. 이 중 10개의 EST에서 insertion/deletion 또는 single nucleotide polymorphism을 발견하여 이를 바탕으로 쉽게 이용이 가능한 10개의 다형성 PCR 마커를 개발하였다. 본 연구에서 개발된 trehalose 분자마커는 앞으로 배추 작물에서 환경 스트레스 저항성과 유전적 연관성을 확인하는데 이용될 수 있고 MAS를 이용한 품종육성에 이용될 수 있다고 기대된다.

Role of Trehalose Synthesis in Ralstonia syzygii subsp. indonesiensis PW1001 in Inducing Hypersensitive Response on Eggplant (Solanum melongena cv. Senryo-nigou)

  • Laili, Nur;Mukaihara, Takafumi;Matsui, Hidenori;Yamamoto, Mikihiro;Noutoshi, Yoshiteru;Toyoda, Kazuhiro;Ichinose, Yuki
    • The Plant Pathology Journal
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    • 제37권6호
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    • pp.566-579
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    • 2021
  • Ralstonia syzygii subsp. indonesiensis (Rsi, former name: Ralstonia solanacearum phylotype IV) PW1001, a causal agent of potato wilt disease, induces hypersensitive response (HR) on its non-host eggplant (Solanum melongena cv. Senryo-nigou). The disaccharide trehalose is involved in abiotic and biotic stress tolerance in many organisms. We found that trehalose is required for eliciting HR on eggplant by plant pathogen Rsi PW1001. In R. solanacearum, it is known that the OtsA/OtsB pathway is the dominant trehalose synthesis pathway, and otsA and otsB encode trehalose-6-phosphate (T6P) synthase and T6P phosphatase, respectively. We generated otsA and otsB mutant strains and found that these mutant strains reduced the bacterial trehalose concentration and HR induction on eggplant leaves compared to wild-type. Trehalose functions intracellularly in Rsi PW1001 because addition of exogenous trehalose did not affect the HR level and ion leakage. Requirement of trehalose in HR induction is not common in R. solanacearum species complex because mutation of otsA in Ralstonia pseudosolanacearum (former name: Ralstonia solanacearum phylotype I) RS1002 did not affect HR on the leaves of its non-host tobacco and wild eggplant Solanum torvum. Further, we also found that each otsA and otsB mutant had reduced ability to grow in a medium containing NaCl and sucrose, indicating that trehalose also has an important role in osmotic stress tolerance.

Synthesis and DNA-binding Properties of Trehalose-tethered Monomeric and Dimeric Berberines

  • Wang, Yong-Min;Zhou, Chun-Qiong;Chen, Jin-Xiang;Chen, Wen-Hua
    • Bulletin of the Korean Chemical Society
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    • 제34권3호
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    • pp.749-752
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    • 2013
  • Trehalose-tethered monomeric and dimeric berberines were synthesized in 50% and 30% from the reaction of berberrubine with 6-tosyl-${\alpha}$,${\alpha}^{\prime}$-trehalose and 6,6'-ditosyl-${\alpha}$,${\alpha}^{\prime}$-trehalose, respectively, and fully characterized by MS (HR and ESI) and NMR ($^1H$, $^{13}C$, COSY and HSQC). Spectrophotometric and spectrofluorimetric titrations indicated that compared with berberine, trehalose-tethered monomeric berberine had comparable DNA-binding affinity toward calf-thymus DNA, whereas trehalose-spaced dimeric berberine exhibited higher DNA-binding affinity. The potential application of these conjugates is also briefly discussed.

Enzymatic Synthesis and Characterization of Galactosyl Trehalose Trisaccharides

  • Kim, Bong-Gwan;Lee, Kyung-Ju;Han, Nam-Soo;Park, Kwan-Hwa;Lee, Soo-Bok
    • Food Science and Biotechnology
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    • 제16권1호
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    • pp.127-132
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    • 2007
  • [ ${\alpha},\;{\alpha}$ ]-Trehalose was efficiently modified by a transgalactosylation reaction of Escherichia coli ${\beta}-galactosidase$ using lactose as a donor to yield two galactosyl trehalose trisaccharides. The reaction products of trehalose by the enzyme were observed by thin layer chromatography (TLC) and high performance anion exchange chromatography (HPAEC) and were purified by BioGel P2 gel permeation chromatography and recycling preparative HPLC. Liquid chromatography-mass spectrometry (LC-MS) and ^{13}C$ nuclear magnetic resonance (NMR) analyses revealed that the structures of the main products were $6^2-{\beta}-D-galactosyl$ trehalose (1) and $4^2-{\beta}-D-galactosyl$ trehalose (2). A reaction of 30%(w/v) trehalose and 15%(w/v) lactose at pH 7.5 and $45^{\circ}C$ resulted in a total yield of approximately 27-30% based on the amount of trehalose used. The galactosyl trehalose products were not hydrolyzed by trehalose. In addition the mixture of transfer products (9:1 ratio of 1 to 2) showed higher thermal stability than glucose, lactose, and maltose, but less than trehalose, against heat treatment over $100^{\circ}C$ at pH 4 and 7. It also exhibited better thermal stability than sucrose at pH 4 alone.

Over-expression of BvMTSH, a fusion gene for maltooligosyltrehalose synthase and maltooligosyltrehalose trehalohydrolase, enhances drought tolerance in transgenic rice

  • Joo, Joungsu;Choi, Hae Jong;Lee, Youn Hab;Lee, Sarah;Lee, Choong Hwan;Kim, Chung Ho;Cheong, Jong-Joo;Choi, Yang Do;Song, Sang Ik
    • BMB Reports
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    • 제47권1호
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    • pp.27-32
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    • 2014
  • Plant abiotic stress tolerance has been modulated by engineering the trehalose synthesis pathway. However, many stress-tolerant plants that have been genetically engineered for the trehalose synthesis pathway also show abnormal development. The metabolic intermediate trehalose 6-phosphate has the potential to cause aberrations in growth. To avoid growth inhibition by trehalose 6-phosphate, we used a gene that encodes a bifunctional in-frame fusion (BvMTSH) of maltooligosyltrehalose synthase (BvMTS) and maltooligosyltrehalose trehalohydrolase (BvMTH) from the nonpathogenic bacterium Brevibacterium helvolum. BvMTS converts maltooligosaccharides into maltooligosyltrehalose and BvMTH releases trehalose. Transgenic rice plants that over-express BvMTSH under the control of the constitutive rice cytochrome c promoter (101MTSH) or the ABA-inducible Ai promoter (105MTSH) show enhanced drought tolerance without growth inhibition. Moreover, 101MTSH and 105MTSH showed an ABA-hyposensitive phenotype in the roots. Our results suggest that over-expression of BvMTSH enhances drought-stress tolerance without any abnormal growth and showes ABA hyposensitive phenotype in the roots.

Construction of Bifunctional Fusion Enzyme between Maltooligosyltrehalose Synthase and Maltooligosyltrehalose Trehalohydrolase of Sulfolobus acidocaldarius and Overexpression in E. coli

  • Kim, Chung Ho
    • 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.

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Bifunctional Recombinant Fusion Enzyme Between Maltooligosyltrehalose Synthase and Maltooligosyltrehalose Trehalohydrolase of Thermophilic Microorganism Metallosphaera hakonensis

  • Seo, Ju-Seok;An, Ju-Hee;Cheong, Jong-Joo;Choi, Yang-Do;Kim, Chung-Ho
    • 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.

Influence of preserved brewing yeast strains on fermentation behavior and flocculation capacity

  • Cheong, Chul;Wackerbauer, Karl;Beckmann, Martin;Kang, Soon-Ah
    • Nutrition Research and Practice
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    • 제1권4호
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    • pp.260-265
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    • 2007
  • Preservation methods on the physiological and brewing technical characters in bottom and top brewing yeast strains were investigated. The preserved yeasts were reactivated after 24 months storage and grown up to stationary phase. The samples of filter paper storage indicated a higher cell growth and viability during propagation than those of nitrogen and lyophilization storage independent on propagation temperature. In addition, the filter paper storage demonstrated a faster absorption of free amino nitrogen and a highest level of higher aliphatic alcohols production during propagation than other preservation methods, which can be attributed to intensive cell growth during propagation. Moreover, the filter paper storage showed a faster accumulation for glycogen and trehalose during propagation, whereas, in particular, lyophilization storage noted a longer adaptation time regarding synthesis of glycogen and trehalose with delayed cell growth. In beer analysis, the filter paper storage formed an increased higher aliphatic alcohols than control. In conclusion, the preservation of filter paper affected positively on yeast growth, viability and beer quality independent on propagation temperature. In addition, in this study, it was obtained that the HICF and Helm-test can be involved as rapid methods for determination of flocculation capacity.

미생물의 세포생리에 미치는 전이방사선의 영향에 관한 연구 (제 4 ) -효모균의 수화물대 에 대한 $\gamma$-의 영향에 대하여- (Studies on the cellular metabolism in microorganisms as influenced by gamma-irradiation.(IV) "on the carbohydrate metabolism of yeast irradiated by $\gamma$-ray.")

  • 김종협
    • 미생물학회지
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    • 제6권2호
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    • pp.41-53
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    • 1968
  • Studies on the carbohydrate metabolism of yeast as influenced by gamma-irradiation from cobalt-60 have been carried, then the mechanisms of radiation effect on respiration and fermentation were discussed under considerations of permeable changes of irradiated cell membrane. The cells of baker's yeast (Saccharomyces cerevisiae) which had been gamma-irradiated of 240 k.r. doses for an hour, then were put into aerobic oxidation and anaerobic fermentation without substrate. Total and fractionated carbohydrates of irradiated yeast cells were determined by calorimetric method with anthrone and orcinol reagents, the amounts of total carbohydrate, trehalose, RNA-ribose, PCA-soluble glycogen, alkali-soluble glycogen, acetic acid-soluble glycogen, mannan and glucan were determined according to the course of aerobic oxidation and anaerobic fermentation. It is found that the carbohydrates of irradiated cells leak out and amount of the losses teaches eleven times more than that of control, the volume of losses are seems to be replaced by water, it can be suggested the damage of gamma-irradiation occurs in the site of passive transport of cell membrane. The endogeneous aerobic respiration of irradiated cells are increased much more than control, the synthesis of reserve glycogen, glucan and RNA-ribose promoted much more than control. The anaerobic fermentation of irradiated cells are also increased than that of control, but the breakdown of carbohydrate is less than endogeneous respiration of irradiated cells. The synthetic rate is also less than that of aerobic oxidation. In irradiated yeast cells, trehalose is revealed to be primary substrate for endogeneous carbohydrate metabolism, so it is proved that the enzymic patterns are not changed but the activities of enzymes relating endogeneous respiration and autofermentation is activated. It is to be considerable to distiguish endogeneous respiration and autofermentation from exogeneous respiration and fermentation on irradiation, for membrane permeability changes and loses out carbohydrate by ionizing radiation.

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