Trehalose Assay Kit

Reference code: K-TREH
SKU: 700004348

100 assays (manual) / 1000 assays (microplate) / 1100 assays (auto-analyser) 

Content: 100 assays (manual) / 1000 assays (microplate) / 1100 assays (auto-analyser)
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 2 years under recommended storage conditions
Analyte: Trehalose
Assay Format: Spectrophotometer, Microplate, Auto-analyser
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Increase
Linear Range: 4 to 80 µg of trehalose per assay
Limit of Detection: 37.5 mg/L
Reaction Time (min): ~ 10 min
Application examples: Honey, mushrooms, bread, beer, seafood (e.g. lobster and shrimp), fruit juices, purees and fillings, nutrition bars, surimi, dehydrated fruits and vegetables, fruit products, white chocolate, sports drinks, dairy products, egg products, soups and sauces, confectionery, chewing gum, cosmetics, pharmaceuticals and other materials (e.g. biological cultures, samples, etc.).
Method recognition: Novel method

The Trehalose assay kit is a simple method for the rapid and reliable measurement and analysis of trehalose in foods, beverages and other materials.

Note for Content: The number of manual tests per kit can be doubled if all volumes are halved.  This can be readily accommodated using the MegaQuantTM  Wave Spectrophotometer (D-MQWAVE).

Browse our full range of monosaccharide and disaccharide test kits.

Scheme-K-TREH TREH Megazyme

Advantages
  • Only enzymatic kit available 
  • Very cost effective 
  • All reagents stable for > 2 years after preparation 
  • Very rapid reaction 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Standard included 
  • Suitable for manual, microplate and auto-analyser formats
Documents
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Validation Report
Publications
Publication

Engineering a high-sugar tolerant strain of Saccharomyces cerevisiae for efficient trehalose production using a cell surface display approach.

Tulsook, K., Bussadee, P., Arnthong, J., Mhuantong, W., Trakarnpaiboon, S., Champreda, V. & Suwannarangsee, S. (2024). Bioresources and Bioprocessing, 11(1), 1-12.

Trehalose production via a one-step enzymatic route using trehalose synthase (TreS) holds significant promise for industrial-scale applications due to its simplicity and utilization of low-cost substrates. However, the development of a robust whole-cell biocatalyst expressing TreS remains crucial for enabling practical and economically viable production. In this study, a high-sugar tolerant strain of S. cerevisiae was screened and employed as a host cell for the cell surface display of TreS from Acidiplasma aeolicum. The resultant strain, S. cerevisiae I3A, exhibited remarkable surface displayed TreS activity of 3358 U/g CDW and achieved approximately 64% trehalose yield (10.8 g/L/h productivity) from maltose. Interestingly, no glucose by-product was observed during trehalose production. The S. cerevisiae I3A cells exhibited reusability for up to 12 cycles leading to potential cost reduction of trehalose products. Therefore, our study demonstrated the development of a high-sugar tolerant S. cerevisiae strain expressing TreS on its surface as a whole-cell biocatalyst for efficient and economical trehalose production with potential applications in the food and pharmaceutical industries.

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Publication

Phenotypic Characterization of Fermentation Performance and Stress Tolerance in Commercial Ale Yeast Strains.

Chen, A., Si, Q., Xu, Q., Pan, C., Cheng, Y. & Chen, J. (2024). Fermentation, 10(7), 364.

Yeast plays a crucial role in the fermentation industry, particularly in alcoholic beverage production, where robustness and metabolic flexibility are essential. This study aimed to investigate the stress tolerance and metabolic capabilities of seven commercial ale yeast strains under various stress conditions, including temperature, pH, osmotic pressure, glucose starvation, and ethanol concentration. Detailed growth assays and stress tolerance tests were utilized to evaluate fermentation efficiency, carbon source utilization, and stress adaptation. Significant variability was observed among the strains. ACY169 and ACY150 demonstrated high overall stress tolerance, making them suitable for high-gravity brewing and processes involving extreme temperature fluctuations. ACY10 showed robust performance under acid stress, making it ideal for sour beer production. In contrast, ACY5 exhibited limited adaptability under stress, with longer doubling times and reduced metabolic activity. The study also revealed differences in carbon source utilization, with ACY169 displaying exceptional metabolic versatility by efficiently fermenting various sugars, including glucose, fructose, maltose, and raffinose. ACY10 and ACY150 exhibited balanced fermentation profiles with high ethanol production rates, while ACY9 demonstrated the highest glucose consumption rate but lower ethanol yields and significant acidification.

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Publication

The characteristics of differentiated yeast subpopulations depend on their lifestyle and available nutrients.

Čáp, M. & Palková, Z. (2024). Scientific Reports, 14(1), 3681.

Yeast populations can undergo diversification during their growth and ageing, leading to the formation of different cell-types. Differentiation into two major subpopulations, differing in cell size and density and exhibiting distinct physiological and metabolic properties, was described in planktonic liquid cultures and in populations of colonies growing on semisolid surfaces. Here, we compare stress resistance, metabolism and expression of marker genes in seven differentiated cell subpopulations emerging during cultivation in liquid fermentative or respiratory media and during colony development on the same type of solid media. The results show that the more-dense cell subpopulations are more stress resistant than the less-dense subpopulations under all cultivation conditions tested. On the other hand, respiratory capacity, enzymatic activities and marker gene expression differed more between subpopulations. These characteristics are more influenced by the lifestyle of the population (colony vs. planktonic cultivation) and the medium composition. Only in the population growing in liquid respiratory medium, two subpopulations do not form as in the other conditions tested, but all cells exhibit a range of characteristics of the more-dense subpopulations. This suggests that signals for cell differentiation may be triggered by prior metabolic reprogramming or by an unknown signal from the structured environment in the colony.

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Publication

Morphological and biochemical responses of a neotropical pest insect to low temperatures.

León-Quinto, T., Madrigal, R., Cabello, E., Fimia, A. & Serna, A. (2024). Journal of Thermal Biology, 119, 103795.

As traditionally cold areas become warmer due to climate change, temperature could no longer be a barrier to the establishment of non-native insects. This is particularly relevant for pest insects from warm and tropical areas, mainly those with some tolerance to moderately low temperatures, which could expand their range into these new locations. From this perspective, in this work we studied the morphological and biochemical responses of the Neotropical pest Paysandisia archon to low temperatures, as part of a possible strategy to colonize new areas. To that end, wild larvae were exposed for 7 days to either low (1 and 5°C) or ambient (23°C) temperatures. We then quantified the inner and outer morphological changes, by X-Ray Computer Tomography and Digital Holographic Microscopy, as well as the accumulation of metabolites acting as potential endogenous cryoprotectants, by Spectrophotometry. We found that Paysandisia archon developed a cold-induced response based on different aspects. On the one hand, morphological changes occurred with a significant reduction both in fluids susceptible to freezing and fat body, together with the thickening, hardening and increased roughness of the integument. On the other hand, we found an increase in the hemolymph concentration of cryoprotective substances such as glucose (6-fold) and glycerol (2-fold), while trehalose remained unchanged. Surprisingly, this species did not show any evidence of cold-induced response unless the environmental temperature was remarkably low (1°C). These results could be useful to improve models predicting the possible spread of such a pest, which should incorporate parameters related to its resistance to low temperatures.

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Publication

Deletion of NTH1 and HSP12 increases the freeze-thaw resistance of baker’s yeast in bread dough.

Chen, B. C. & Lin, H. Y. (2022). Microbial Cell Factories, 21(1), 1-10.

Background: The intracellular molecule trehalose in Saccharomyces cerevisiae may have a major protective function under extreme environmental conditions. NTH1 is one gene which expresses trehalase to degrade trehalose. Small heat shock protein 12 (HSP12 expressed) plays a role in protecting membranes and enhancing freezing stress tolerance. Results: An optimized S. cerevisiae CRISPR-Cpf1 genome-editing system was constructed. Multiplex genome editing using a single crRNA array was shown to be functional. NTH1 or/and HSP12 knockout in S. cerevisiae enhanced the freezing stress tolerance and improved the leavening ability after freezing and thawing. Conclusions: Deleting NTH1 in the combination with deleting HSP12 would strengthen the freezing tolerance and protect the cell viability from high rates of death in longer-term freezing. It provides valuable insights for breeding novel S. cerevisiae strains for the baking industry through a more precise, speedy, and economic genome-editing system.

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Publication

Increased Stress Levels in Caged Honeybee (Apis mellifera)(Hymenoptera: Apidae) Workers.

Lattorff, H. M. G. (2022). Stresses, 2(4), 373-383.

Honeybees, Apis mellifera, usually live in large colonies consisting of thousands of individuals. Within the colony, workers interact with their social environment frequently. The large workforce, division of labour, and other features may promote the ecological success of honeybees. For decades, artificial mini colonies in cages within the laboratory have become the gold standard, especially in experiments related to toxicology, effects of pesticides and pathogens. Experiments using caged bees and full-sized colonies yielded contradictory results. Here, the effect of cage experiments on the stress level of individual bees is analysed. Two different stress response were targeted, the heat shock response and the mobilization of energetic resources. While no differences were found for varying group sizes of bees, very strong effects emerged by comparing caged workers with bees from natural colonies. Caged workers showed increased levels of hsp expression and reduced haemolymph titres for trehalose, the energy storage sugar. These results reveal that the lack of the social environment (e.g., lack of queen, lack of sufficient group size) induce stress in caged bees, which might act synergistically when bees are challenged by additional stressors (e.g., pesticides, pathogens) resulting in higher mortality than observed under field conditions.

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Publication

Chronic exposure to field-realistic doses of imidacloprid resulted in biphasic negative effects on honey bee physiology.

Kim, S., Kim, J. H., Cho, S., Clark, J. M. & Lee, S. H. (2022). Insect Biochemistry and Molecular Biology, 144, 103759.

There have been many investigations on the negative effects of imidacloprid (IMD) on honey bees. IMD is known to disrupt honey bee physiology and colony health at a relatively low concentration compared to other pesticides. In this study, honey bee colonies were chronically exposed to field-realistic concentrations (5, 20, and 100 ppb) of IMD, and the body weight, flight performance, carbohydrate reserve, and lipid contents of forager bees analyzed. Transcriptome analyses followed by quantitative PCR were also conducted for both nurse and forager bees to elucidate any changes in energy metabolism related to phenotypic disorders. The body weights of newly emerged and nurse bees showed decreasing tendencies as the IMD concentration increased. In forager bees, however, IMD induced a biphasic change in body weight: body weight was decreased at the lower concentrations (5 and 20 ppb) but increased at the higher concentration (100 ppb). Nevertheless, the flight capability of forager bees significantly decreased in a concentration-dependent manner. The effects of IMD on target gene transcription in forager bees showed biphasic patterns between low (5 and 20 ppb) and high (100 ppb) concentrations. Nurse bees showed typical features of premature transition to foragers in a concentration-dependent manner. When exposed to low concentrations, forager bees exhibited downregulation of genes involved in carbohydrate and lipid metabolism and in the insulin/insulin-like growth factor signaling pathway, upregulation of transporter activity, and a dose-dependent body weight reduction, which were similar to insulin resistance and diabetic symptoms. However, increased lipid metabolism and decreased energy metabolism with body weight gain were observed at high IMD concentration. Considered together, these results suggest that field-realistic doses of IMD alter honey bee energy metabolism in distinctly different ways at low and high concentrations, both of which negatively affect honey bee colony health.

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Publication

Effects of Water Stress on Quality and Sugar Metabolism in ‘Gala’Apple Fruit.

Tao, H., Sun, H., Wang, Y., Wang, X. & Guo, Y. (2022). Horticultural Plant Journal, In Press.

Sugar plays an important role in apple fruit development, appearance and quality as well as contributing to a plant's water stress response. Trehalose and the trehalose biosynthetic metabolic pathways are part of the sugar signaling system in plants, which are important regulator of water stress response in apple. The effect of water stress treatments applied to apple trees and the corresponding effects of ABA on developmental fruit quality were examined for indicators of fruit quality during fruit development. The results indicated that the severe water stress treatment (W2) occurring after the last stage of fruit cell division caused a decrease in the color and size of fruit. The moderate water stress (W1) occurring after the last stage of fruit cell enlargement (S2) caused an increase in the content of fructose and sorbitol while the apple fruit shape was not affected. These changes in sugar are related to the activity of sugar metabolic enzymes. While the enzymatic activity of vacuolar acid invertase (vAINV) was higher, that of sucrose-phosphate synthase (SPS) was lower in water stress treated fruit throughout the developmental period. This indicates that enhanced sucrose degradation and reduced sucrose synthesis leads to an overall reduced sucrose content during times of drought. Thus, water stress reduced sucrose content. Whereas the content of endogenous trehalose and ABA were the highest in water stress treated fruit. A moderate water stress (W1) imposed on apple trees via water restriction (60%–65% of field capacity) after the fruit cell enlargement phase of fruit development yielded sweeter fruit of higher economic value.

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Publication

Trehalose provisioning in Daphnia resting stages reflects local adaptation to the harshness of diapause conditions.

Santos, J. L. & Ebert, D. (2022). Biology Letters, 18(2), 20210615.

Environmental fluctuations often select for adaptations such as diapause states, allowing species to outlive harsh conditions. The natural sugar trehalose which provides both cryo- and desiccation-protection, has been found in diapause stages of diverse taxa. Here, we hypothesize that trehalose deposition in resting stages is a locally adapted trait, with higher concentrations produced in harsher habitats. We used resting stages, produced under standardized conditions, by 37 genotypes of Daphnia magna collected from Western Palaearctic habitats varying in their propensity to dry in summer and freeze in winter. Resting eggs produced by D. magna from populations from summer-dry habitats showed significantly higher trehalose than those from summer-wet habitats, suggesting that trehalose has a protective function during desiccation. By contrast, winter-freezing did not explain variation in trehalose content. Adaptations to droughts are important, as summer dryness of water bodies is foreseen to increase with ongoing climate change.

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Publication

The photosynthetic efficiency and carbohydrates responses of six edamame (Glycine max. L. Merrill) cultivars under drought stress.

Hlahla, J. M., Mafa, M. S., Van der Merwe, R., Alexander, O., Duvenhage, M. M., Kemp, G. & Moloi, M. J. (2022). Plants, 11(3), 394.

Vegetable-type soybean, also known as edamame, was recently introduced to South Africa. However, there is lack of information on its responses to drought. The aim of this study was to investigate the photosynthetic efficiency and carbohydrates responses of six edamame cultivars under drought stress. Photosynthetic efficiency parameters, including chlorophyll fluorescence and stomatal conductance, were determined using non-invasive methods, while pigments were quantified spectrophotometrically. Non-structural carbohydrates were quantified using Megazyme kits. Structural carbohydrates were determined using Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Drought stress significantly increased the Fv/Fm and PIabs of AGS429 and UVE17 at pod filling stage. Chlorophyll-a, which was most sensitive to drought, was significantly reduced in AGS429 and UVE17, but chlorophyll-b was relatively stable in all cultivars, except UVE17, which showed a significant decline at flowering stage. AGS354 and AGS429 also showed reduced chlorophyll-b at pod filling. UVE17 showed a significant reduction in carotenoid content and a substantial reduction in stomatal conductance during pod filling. Drought stress during pod filling resulted in a significant increase in the contents of trehalose, sucrose and starch, but glucose was decreased. Chlorophyll-a positively correlated with starch. The FTIR and XRD results suggest that the cell wall of UVE14, followed by UVE8 and AGS429, was the most intact during drought stress. It was concluded that carotenoids, stomatal conductance, starch and hemicellulose could be used as physiological/biochemical indicators of drought tolerance in edamame. This information expands our knowledge of the drought defense responses in edamame, and it is essential for the physiological and biochemical screening of drought tolerance.

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Publication

Roles of PINK1 in regulation of systemic growth inhibition induced by mutations of PTEN in Drosophila.

Han, Y., Zhuang, N., & Wang, T. (2021). Cell Reports, 34(12), 108875.

The maintenance of mitochondrial homeostasis requires PTEN-induced kinase 1 (PINK1)-dependent mitophagy, and mutations in PINK1 are associated with Parkinson’s disease (PD). PINK1 is also downregulated in tumor cells with PTEN mutations. However, there is limited information concerning the role of PINK1 in tissue growth and tumorigenesis. Here, we show that the loss of pink1 caused multiple growth defects independent of its pathological target, Parkin. Moreover, knocking down pink1 in muscle cells induced hyperglycemia and limited systemic organismal growth by the induction of Imaginal morphogenesis protein-Late 2 (ImpL2). Similarly, disrupting PTEN activity in multiple tissues impaired systemic growth by reducing pink1 expression, resembling wasting-like syndrome in cancer patients. Furthermore, the re-expression of PINK1 fully rescued defects in carbohydrate metabolism and systemic growth induced by the tissue-specific pten mutations. Our data suggest a function for PINK1 in regulating systemic growth in Drosophila and shed light on its role in wasting in the context of PTEN mutations.

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Sulfolobus acidocaldarius uses a complex trehalose metabolism for salt stress response involving a novel TPS/TPP pathway.

Stracke, C., Meyer, B. H., Hagemann, A., Jo, E., Lee, A., Albers, S. V., Cha, J., Brasen, C. & Siebers, B. (2020). Applied and Environmental Microbiology, 86(24).

The crenarchaeon Sulfolobus acidocaldarius has been described to synthesize trehalose via the maltooligosyltrehalose synthase (TreY) and maltooligosyltrehalose trehalohydrolase (TreZ) pathway, and the trehalose glycosyltransferring synthase (TreT) pathway has been predicted. Deletion mutant analysis of strains with single and double deletions of ΔtreY and ΔtreT in S. acidocaldarius revealed that in addition to these two pathways, a third, novel trehalose biosynthesis pathway is operative in vivo: the trehalose-6-phosphate (T6P) synthase/T6P phosphatase (TPS/TPP) pathway. In contrast to known TPS proteins, which belong to the GT20 family, the S. acidocaldarius TPS belongs to the GT4 family, establishing a new function within this group of enzymes. This novel GT4-like TPS was found to be present mainly in the Sulfolobales. The ΔtreY ΔtreT Δtps triple mutant of S. acidocaldarius, which lacks the ability to synthesize trehalose, showed no altered phenotype under standard conditions or heat stress but was unable to grow under salt stress. Accordingly, in the wild-type strain, a significant increase of intracellular trehalose formation was observed under salt stress. Quantitative real-time PCR showed a salt stress-mediated induction of all three trehalose-synthesizing pathways. This demonstrates that in Archaea, trehalose plays an essential role for growth under high-salt conditions.

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Engineering cyanobacteria as cell factories for direct trehalose production from CO2.

Qiao, Y., Wang, W. & Lu, X. (2020). Metabolic Engineering, 62, 161-171.

Trehalose is a non-reducing disaccharide with a wide range of applications in food, cosmetic, and pharmaceutical industries. Cyanobacteria are promising cell factories to produce biochemicals by using solar energy and CO2. Trehalose is biosynthesized at low intracellular concentrations as a salt-inducible compatible solute in some cyanobacteria. In the current study, we demonstrated the efficient trehalose production without salt induction in cyanobacteria by metabolic engineering. The trehalose transporter 1 (TRET1) from an anhydrobiotic insect (Polypedilum vanderplanki) was successfully expressed in the engineered strains and the intracellular trehalose was efficiently secreted to the medium. As the results, the engineered strain co-expressing maltooligosyl trehalose synthase (MTS), maltooligosyl trehalose trehalohydrolase (MTH) and TRET1 secreted 97% of trehalose to the medium, and the titer was up to 2.7 g/L in 15 days. In addition, 5.7 g/L trehalose was produced by semi-continuous cultivation in 34 days. Taken together, this work demonstrates cyanobacteria can be applied as cell factories for direct sunlight-driven conversion of CO2 into excreted trehalose.

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The effect of trehalose on autophagy-related proteins and cyst growth in a hypomorphic Pkd1 mouse model of autosomal dominant polycystic kidney disease.

Atwood, D. J., Brown, C. N., Holditch, S. J., Pokhrel, D., Thorburn, A., Hopp, K. & Edelstein, C. L. (2020). Cellular Signalling, 75, 109760.

Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disorder characterized by kidney cyst growth often resulting in end-stage renal disease. There is growing attention on understanding the role of impaired autophagy in ADPKD. Trehalose (TRE) has been shown to increase both protein stability and aggregate clearance and induce autophagy in neurodegenerative diseases. TRE treatment in wild type mice compared to vehicle resulted in increased expression in the kidney of Atg12-5 complex and increased Rab9a, autophagy-related proteins that play a role in the formation of autophagosomes. Thus, the aim of the study was to determine the effect of TRE on cyst growth and autophagy-related proteins, in the hypomorphic Pkd1RC/RC mouse model of ADPKD. Pkd1RC/RC mice were treated 2% TRE in water from days 50 to 120 of age. TRE did not slow cyst growth or improve kidney function or affect proliferation and apoptosis in Pkd1RC/RC kidneys. In Pkd1RC/RC vs. wild type kidneys, expression of the Atg12-5 complex was inhibited by TRE resulting in increased free Atg12 and TRE was unable to rescue the deficiency of the Atg12-5 complex. Rab9a was decreased in Pkd1RC/RC vs. wild type kidneys and unaffected by TRE. The TRE-induced increase in p62, a marker of autophagic cargo, that was seen in normal kidneys was blocked in Pkd1RC/RC kidneys. In summary, the autophagy phenotype in Pkd1RC/RC kidneys was characterized by decreases in crucial autophagy-related proteins (Atg12-5 complex, Atg5, Atg16L1), decreased Rab9a and increased mTORC1 (pS6S240/244, pmTORS2448) proteins. TRE increased Atg12-5 complex, Rab9a and p62 in normal kidneys, but was unable to rescue the deficiency in autophagy proteins or suppress mTORC1 in Pkd1RC/RC kidneys and did not protect against cyst growth.

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Combination of hydrophobically modified γ-poly (glutamic acid) and trehalose achieving high cryosurvival of RBCs.

Zhang, Q., Liu, B., Chong, J., Ren, L., Zhao, Y. & Yuan, X. (2020). Science China Technological Sciences, 1-11.

Trehalose is expected to be an alternative for toxic glycerol as a biocompatible cryoprotectant of red blood cells (RBCs). In this work, γ-poly(glutamic acid) (PGA) is modified by grafting hydrophobic phenethylamine, 3,4-dimethoxyphenylethylamine and dodecylamine, respectively. The graft-modified PGA can significantly enhance cryosurvival of RBCs in combination with trehalose. Analyses of dynamic light scattering, hemolysis assay, atomic force microscope and confocal laser scanning microscope suggest that the modified PGA polymers can self-assemble into nanoparticles in phosphate buffer saline solutions at the pH range of 6.0-7.4, and exhibit membrane-disruptive activity due to hydrogen bond, conjugation and hydrophobic interactions with cell membranes. It is assumed that the modified PGA polymers can improve the cryosurvival of RBCs by promoting membrane permeability of trehalose. Among the three graft-modified polymers, phenethylamine-grafted PGA (PGA-g-PEA) can significantly increase the intracellular trehalose-loading content to 11.3±2.4 mM at pH 7.4, much higher than the value 0.17±0.66 mM when trehalose is used without any polymers. In view of the aforementioned merit, the cryosurvival rate of sheep RBCs is increased to about 90% by incubation with 1.0 mg mL-1 PGA-g-PEA and 0.36 M trehalose. In vitro cell culture of L929 fibroblasts demonstrates low cytotoxicity of PGA-g-PEA. Therefore, hydrophobic PEA-modified PGA with enhanced intracellular trehalose-loading ability can be potentially applied in glycerol-free RBC cryopreservation or other related biomacromolecule delivery systems.

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Safety Information
Symbol : GHS05, GHS08
Signal Word : Danger
Hazard Statements : H314, H360
Precautionary Statements : P201, P202, P260, P264, P280
Safety Data Sheet
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