• Title/Summary/Keyword: Trehalose

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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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    • v.16 no.1
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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.

Trehalose가 발광미생물의 동결건조시 생존 및 발광강도에 미치는 영향과 첨가방법에 대한 연구

  • Park, Ji-Eun;Jang, Deok-Jin
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.390-393
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    • 2000
  • In this work, effects of trehalose on the recovery of bioluminescence and viability of luxAB-containing recombinant bacteria were investigated. In case of a recombinant, UV2, only 2.5% of bioluminescence and 2.7% of cell viability were restored after 3.5hr of freeze-drying without trehalose, which implies that cells were heavily damaged during the dehydration. To improve these losses, trehalose was added before freeze-drying on different modes. Trehalose increased the bioluminescence and viability of freeze-dried UV2 in all conditions tested and it was observed that addition of trehalose into the broth(final concentraion, 0.08M) for 15min before the freeze-drying resulted in 45% of bioluminescence and 50% of cell viability. For the other luminescent recombinant, YH9, trehalose showed a similar efficacy.

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Effect of Trehalose Accumulation on the Intrinsic and Acquired Thermotolerance in a Natural Isolate, Saccharomyces cerevisiae KNU5377

  • PAIK, SANG-KYOO;HAE-SUN YUN;HO-YONG SOHN;INGNYOL JIN
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.85-89
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    • 2003
  • The difference in the thermotolerance between Saccharomyces cerevisiae KNU5377 and ATCC24858 was compared by assaying the amounts of trehalose accumulated under growth and heat shock conditions. Both strains exhibited similar trehalose accumulation during the growth period, but an intrinsic thermotolerance was much higher in KNU5377 than in the control strain. This result implied that some strain-specific characteristics of KNU5377, other than trehalose accumulation, primarily were responsible fur its higher intrinsic thermotolerance. Heat shock at $43^{\circ}C$ for 90 min to the exponentially growing cells resulted in the maximum level of trehalose In both strains. Trehalose accumulated at least twice more in KNU5377 by the heat shock than in the control, due to the maintenance of its neutral trehalase activity even after the heat shock. Consequently, the Increase of acquired thermotolerance in both strains correlated with an increase in the trehalose content in each strain. In conclusion, KNU5377 exhibited a well-modulated trehalose-related mechanism to accumulate more trehalose by means of maintaining neutral trehalase activity after heat shock than the control strain, thereby contributing to its acquired thermotolerance.

동결 건조법을 이용한 레트로바이러스 저장의 안정화에 미치는 trehalose의 영향

  • Jo, Su-Hyeong;Kim, Byeong-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.309-312
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    • 2000
  • We investigated the effect of various additives on the freeze-drying efficiency of recombinant retrovirus. Trehalose was the more effective protectant compared with sucrose or glucose and the optimize trehalose concentration was 300 mM. To improve the effect of trehalose, trehalose-polymers mixtures were tested as protectants so that Poly(vinylpyrollidon)(PVP) improved the recovery yield slightly, but dextran and Poly(ethylenglycol)(PEG) showed a negative effect.

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Production of Recombinant Trehalose Synthase from Thermus caldophilus GK24 (재조합 내열성 트레할로스 합성효소의 생산)

  • Choi, Jae-Youl;Cha, Wol-Suk;Shin, Hyun-Jae
    • KSBB Journal
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    • v.21 no.4
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    • pp.298-301
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    • 2006
  • A gene(GeneBank AF 135796) coding for a trehalose synthase from Thermus caldophilus GK24 was cloned into Escherichia coli K12 using five vector systems. The constitutive expression system(pHCETS) which shows the highest trehalose synthase activity from flask culture of recombinant E. coli was selected for the production of trehalose from maltose. For the shake flask culture, the final dry cell weight was 0.9 g/L and the trehalose synthase activity was 25 U/mL. Fed-batch culture of recombinant E. coli harboring plasmid pHCETS which uses the glycerolas a carbon source was performed in jar fermentor: the dry cell weight of 20 g/L and the trehalose synthase activity of 13.7 U/mL were attained in 48 h.

Physical Properties and Sensory Evaluation of Muffins with Trehalose (트레할로스를 첨가한 머핀의 물리적 특성 및 관능평가)

  • Heo, Soo-Jin;An, Hye-Lyung;Lee, Kwang-Suck
    • Culinary science and hospitality research
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    • v.16 no.1
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    • pp.13-23
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    • 2010
  • The principal objective of this study was to develop the optimal recipe for muffins prepared with replacement of sucrose with trehalose. The effects of trehalose on properties and staling of muffins during storage days(0, 1, 3, 5 days) were evaluated in terms of height, volume, weight, specific volume, baking loss rate, crumbscan, colorimeter, texture analyzer and sensory evaluation. Crust thickness of muffins containing trehalose evaluated with crumbscan decreased as the content of trehalose increased. Lightness(L value) of muffins with trehalose increased for the storage days, but muffins without trehalose decreased. yellowness(b value) increased significantly as the trehalose content increased. Hardness value of muffins was reduced by adding trehalose; however, the resilience value of muffins with trehalose increased significantly. Finally, the sensory evaluation revealed that muffins with 25% of trehalose showed the best result in texture, taste and overall preference.

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Effect of Trehalose on Bioluminescence and Viability of Freeze-Dried Bacterial Cells

  • PARK, JI-EUN;KYU-HO LEE;DEOKJIN JAHNG
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.349-353
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    • 2002
  • Two recombinant bacteria containing luxAB showed an increased tolerance to stresses associated with lyophilization, when the cells were freeze-dried in the presence of trehalose. In the case of a recombinant, UV2, only $2.5\%$ of the original bioluminescence and $2.7\%$ of the cell viability were restored after 4 h of freeze-drying without trehalose, which implies that the cells were heavily damaged during the dehydration. To improve these losses, trehalose was added before freeze-drying using different modes. Trehalose increased the bioluminescence and the viability of freeze-dried UV2 under all conditions tested, and it was also observed that the addition of trehalose to the cultures (final concentration of 0.08 M) for 15 min before the freeze-drying resulted in the restoration of $45\%$ of the original bioluminescence and $50\%$ of the cell viability. Trehalose also showed a similar efficacy with the other luminescent recombinant, YH9. Therefore, it was tentatively concluded that trehalose played a role as a protective agent in the freeze-drying of bacterial cells.

In Vitro Development of Porcine Oocytes Following Intracytoplasmic Injection of Freeze-Dried Spermatozoa with Trehalose (Trehalose에 의하여 동결 건조된 정자의 돼지 난자 내 직접주입 후 체외 배발달)

  • Kang, Hwa-Hyung;Lee, Ji-Woong;Kang, Man-Jong;Kim, Kwang-Hyun;Moon, Seung-Ju
    • Journal of Embryo Transfer
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    • v.29 no.1
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    • pp.51-57
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    • 2014
  • The objectives of this study were to investigate the effects of trehalose as a cryoprotectant for porcine freeze-dried spermatozoa, to find the optimal freeze-drying time and storage periods of freeze-dried spermatozoa, and to find out pronuclear formation rates, cleaved rates, and embryo development through intracytoplasmic injection of freeze-dried spermatozoa on porcine oocytes. The survival rates of spermatozoa after freeze-drying with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). The highest survival rates were found at 75 mM trehalose treatment. The longer storage periods after freeze-drying seemed to have a lower survival rates. Development in culture of pig by ICSI with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). Shorter freeze-drying time of spermatozoa was resulted in the highest cleaved rates and embryo development.

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

  • Hong, Jai-Sik;Yun, Se-Eok;Kim, Young-Soo;Lee, Jong-Bae
    • The Korean Journal of Mycology
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    • v.15 no.2
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    • pp.108-115
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    • 1987
  • Nutritional characteristics and cultural conditions for trehalose synthesis and mycelial yield of Pleurotus sajor-caju and Pleurotus ostreatus 201 in submerged culture were investigated. The results were as follows: Among the carbon sources, glucose was most excellent for trehalose synthesis and mycelial yield. The optimal concentration of glucose was 1%. Among the nitrogen sources, peptone was most excellent for trehalose synthesis and mycelial yield. The optimal concentration of peptone was 0.05%. The optimal concentration of $KH_2PO_4$ and $MgSO_47H_2O$ for trehalose synthesis and mycelial yield was 0.1%,0.04% and 0.2%,0.04-0.08%, respective­ly. The optimal temperature and pH for trehalose synthesis were $25^{\circ}C$ and pH 5.5. but optimal temperature and pH for mycelial yield were $30^{\circ}C$ and pH 5.5. The maximum yield of trehalose was obtanined after 10 day cultivation.

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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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    • v.37 no.6
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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.