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Whole-genome duplication (WGD) plays important roles in plant evolution and function, yet little is known about how WGD underlies metabolic diversification of natural products that bear significant medicinal properties, especially in nonmodel trees. Here, we reveal how WGD laid the foundation for co-option and differentiation of medicinally important ursane triterpene pathway duplicates, generating distinct chemotypes between species and between developmental stages in the apple tribe. After generating chromosome-level assemblies of a widely cultivated loquat variety and Gillenia trifoliata, we define differentially evolved, duplicated gene pathways and date the WGD in the apple tribe at 13.5 to 27.1 Mya, much more recent than previously thought. We then functionally characterize contrasting metabolic pathways responsible for major triterpene biosynthesis in G. trifoliata and loquat, which pre- and postdate the Maleae WGD, respectively. Our work mechanistically details the metabolic diversity that arose post-WGD and provides insights into the genomic basis of medicinal properties of loquat, which has been used in both traditional and modern medicines.
Assuntos
Eriobotrya/genética , Duplicação Gênica , Poliploidia , Triterpenos/metabolismo , Vias Biossintéticas , Eriobotrya/metabolismo , Genoma de PlantaRESUMO
A sensitive chemiluminescent enzyme immunoassay (CLEIA) was established for the determination of gentamicin (GEN) residue levels in animal tissue. This assay is based on a fusion protein of single-chain variable fragment (scFv) and alkaline phosphatase (AP). Initially, VL and VH derived from anti-gentamicin monoclonal antibody were linked by a short peptide to construct a scFv. Subsequently, the constructed scFv sequence was accessed into the pLIP6/GN vector, and a soluble scFv-AP fusion protein was generated. The scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) for the determination of gentamicin. In the dcCLEIA, the half inhibitory concentration (IC50) and limit of detection (LOD) were 1.073 ng/mL and 0.380 ng/mL, respectively. The average recoveries of gentamicin spiked in animal tissue samples ranged from 78% to 96%. These results showed a strong correlation with ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The above results suggest that the anti-GEN scFv-AP fusion protein is suitable for detecting gentamicin residues in edible animal tissues.
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The aim of this study is to develop a simple but rapid method for the determination of foodborne pathogens in complex matrices (beverages) by surface enhanced Raman spectroscopy (SERS) combined with Au nanostar solid-phase substrates. The star-shaped singlet Au nanostructure was formed on the surface of a stainless steel sheet by chemical replacement reaction. Rhodamine 6G verified the sensitivity and reproducibility of this substrate, and the relative standard deviations of the SERS intensity at 1312 cm-1, 1364 cm-1, and 1510 cm-1 displacements were 3.40%, 5.64%, and 3.48%, respectively. By detecting four pathogens in beverage samples on Au nanostar substrates, the utility of the SERS assay was demonstrated, while the combination of principal component analysis (PCA) and hierarchical cluster analysis (HCA) further enabled the isolation and identification of pathogens. The results of spiked beverages were validated in conventional culture identification and Vitek 2 Compact biochemical identification system experiments. Thus, this research demonstrated that Au nanostar substrates can be effectively utilized for the recognition of pathogenic bacteria and have immense promise to advance the progress of quick detection of foodborne pathogens and food safety.
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Nanopartículas Metálicas , Nanopartículas Metálicas/química , Reprodutibilidade dos Testes , Ouro/química , Análise Espectral Raman/métodos , BebidasRESUMO
Electrochemical sensor has great potential in the detection of small molecules by virtues of low cost, fast response, and easy to miniaturization. However, electrochemical sensing of ochratoxin A (OTA) was seriously hindered by the heavy electrode-fouling effect and poor electrochemical activity inherent from OTA molecular. Herein, two-dimensional titanium carbide (2D Ti3C2) MXene incorporated with carboxylic multiwalled carbon nanotubes (cMWCNTs) was developed as a glassy carbon electrode modifier for rapid and sensitive detection of OTA. Physical characterizations combined with electrochemical techniques revealed that cMWCNTs can not only prevent the restacking of 2D Ti3C2nanosheets but also facile its electron transfer, leading to a nanohybrid with a high specific surface and good electrocatalytic activity to OTA. Under optimal conditions, the electrochemical sensor showed a good linear response to OTA in a concentration range from 0.09 to 10µmol·l-1and a low detection limit (LOD) of 0.028µmol·l-1. The proposed sensor was impelled successive times to detect OTA, a good repeatability was obtained, indicating the constructed sensor possessed good anti-fouling property. Moreover, satisfactory recoveries between 91.8% and 103.2% were obtained in the real sample analysis of grape and beer, showing that the developed sensing technique is reliable for the screening of trace OTA in food resources.
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Técnicas Biossensoriais/instrumentação , Nanotubos de Carbono/química , Ocratoxinas/análise , Titânio/química , Incrustação Biológica , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/instrumentação , Limite de DetecçãoRESUMO
Eighteen derivatives of pentacyclic triterpene carboxylic acids (Maslinic acid, Corosolic acid and Asiatic acid) have been prepared by coupling the piperazine complex of l-amino acids at the C-28 site of the parent compounds. The α-glucosidase inhibitory activities of the pristine derivatives were evaluated in vitro. The results indicated that the inhibitory activity of some compounds (15e IC50 = 591 µM, 16e IC50 = 423 µM) was closed to that of the reference acarbose (IC50 = 347 µM) in ethanol-water system. In addition, compound 16e (IC50 = 380 µM) showed superior inhibitory activity than acarbose (IC50 = 493 µM) in the measurement system with DMSO as solvent. The comparison of two different solvent systems showed that the derivatives had better α-glucosidase inhibitory activity in the DMSO system than that of in ethanol-water system. Regrettably, all of the as-synthesized derivatives exhibited inferior α-glucosidase inhibitory activities than those of the parent compounds in both test solvent systems. Furthermore, the result of enzyme kinetics demonstrated that the inhibition mechanism of compound 16e was noncompetitive inhibition with the inhibition constant Ki = 552 µM.
Assuntos
Aminoácidos/farmacologia , Ácidos Carboxílicos/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Triterpenos Pentacíclicos/farmacologia , Piperazina/farmacologia , alfa-Glucosidases/metabolismo , Aminoácidos/química , Ácidos Carboxílicos/síntese química , Ácidos Carboxílicos/química , Relação Dose-Resposta a Droga , Inibidores de Glicosídeo Hidrolases/síntese química , Inibidores de Glicosídeo Hidrolases/química , Humanos , Estrutura Molecular , Triterpenos Pentacíclicos/síntese química , Triterpenos Pentacíclicos/química , Piperazina/química , Relação Estrutura-AtividadeRESUMO
A novel electrochemical sensor has been fabricated for sensitive determination of zearalenone (ZEA) in food samples based on molybdenum disulfide quantum dots (MoS2 QDs) and two-dimensional titanium carbide (2D-Ti3C2Tx MXene) co-modified multi-walled carbon nanotube (MWCNT) heterostructure. Physical and electrochemical characterizations reveal that 2D-Ti3C2Tx and MoS2 QDs co-modified MWCNTs yields synergistic signal amplification effect, together with large specific surface area and excellent conductivity for the heterostructure, endowing the developed sensor with high detection performance to ZEA. Under optimized conditions, the sensor shows a wide linear range from 3.00 to 300 ng mL-1 and a low limit of detection (LOD) of 0.32 ng mL-1, which is far lower than the maximum residue limits (MRLs) settled by the European Commission. In addition, it exhibits excellent selectivity, high reproducibility with a relative standard deviation (RSD) of 1.1%, and good repeatability (RSD 1.1%). In real sample analysis recoveries ranged from 94.8 to 105% showing the proposed electrochemical sensor has high potential in practical applications. This work presents an effective and valuable pathway for the use of novel heterostructure in the biosensing field.
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Dissulfetos/química , Técnicas Eletroquímicas/métodos , Molibdênio/química , Nanotubos de Carbono/química , Pontos Quânticos/química , Titânio/química , Zearalenona/análise , Técnicas Eletroquímicas/instrumentação , Eletrodos , Farinha/análise , Contaminação de Alimentos/análise , Limite de Detecção , Reprodutibilidade dos Testes , Zea mays/químicaRESUMO
Objectives: Colorectal cancer (CRC) screening using stool samples is now in routine use where tumor DNA methylation analysis for leading markers such as NDRG4 and SDC2 is an integral part of the test. However, processing stool samples for reproducible and efficient extraction of human genomic DNA remains a bottleneck for further research into better biomarkers and assays. Methods: We systematically evaluated several factors involved in the processing of stool samples and extraction of DNA. These factors include: stool processing (solid and homogenized samples), preparation of DNA from supernatant and pellets, and DNA extraction with column and magnetic beads-based methods. Furthermore, SDC2 and NDRG4 methylation levels were used to evaluate the clinical performance of the optimal protocol. Results: The yield of total and human genomic DNA (hgDNA) was not reproducible when solid stool scraping is used, possibly due to sampling variations. More reproducible results were obtained from homogenized stool samples. Magnetic beads-based DNA extraction using the supernatant from the homogenized stool was chosen for further analysis due to better reproducibility, higher hgDNA yield, lower non-hgDNA background, and the potential for automation. With this protocol, a combination of SDC2 and NDRG4 methylation signals with a linear regression model achieved a sensitivity and specificity of 81.82 and 93.75%, respectively. Conclusions: Through the systematic evaluation of different stool processing and DNA extraction methods, we established a reproducible protocol for analyzing tumor DNA methylation markers in stool samples for colorectal cancer screening.
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Biomarcadores Tumorais/análise , Neoplasias Colorretais/diagnóstico , DNA/análise , Testes Diagnósticos de Rotina/métodos , Detecção Precoce de Câncer/métodos , Fezes/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/química , DNA/química , Metilação de DNA , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Musculares/genética , Proteínas do Tecido Nervoso/genética , Reprodutibilidade dos Testes , Manejo de Espécimes/métodos , Sindecana-2/genéticaRESUMO
Combining two bioactive moieties by covalent bond into a novel single hybrid biological entity in view of the principle of active splicing, twenty-two C28-modified derivatives of pentacyclic dihydroxytriterpene carboxylic acids with saturated nitrogen heterocycle segments (i.e. 1-deoxynojirimycin or piperazines) have been synthesized. The inhibitory activities of all final target compounds on α-glucosidase were evaluated in vitro. The results of α-glucosidase inhibition assay indicate that some derivatives (e.g. 4b: IC50 = 1468.4 µM; 12b: IC50 = 499.6 µM 12c: IC50 = 768.5 µM, 13c: IC50 = 819.2 µM) show superior inhibitory activity in α-glucosidase than that of the precursor maslinic acid (IC50 = 2540.6 µM) or corosolic acid (IC50 = 1363.7 µM), in which compound 12b (IC50 = 499.6 µM) possesses stronger inhibitory activity than that of acarbose (IC50 = 606 µM). In addition, the result of enzyme kinetics study reveals that the inhibitory mechanism of the compound 12b is non-competitive inhibition and the inhibition constant Ki is 570 µM. The binding interaction between compounds with α-glucosidase are predicted by molecular docking simulation.
Assuntos
Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/farmacologia , Triterpenos/química , Triterpenos/farmacologia , Técnicas de Química Sintética , Inibidores de Glicosídeo Hidrolases/síntese química , Humanos , Modelos Moleculares , Simulação de Acoplamento Molecular , Saccharomyces cerevisiae/enzimologia , Triterpenos/síntese química , alfa-Glucosidases/metabolismoRESUMO
Twenty maslinic acid and corosolic acid derivatives were obtained by coupling with l-amino acids at C-28 position. The α-glucosidase inhibitory activities of the present compounds were evaluated in vitro. Results reveal that some of the derivatives exhibit a better α-glucosidase inhibitory activity than that of acarbose in the test conditions of ethanol-water solution and DMSO. It is worth noting that maslinic acid and corosolic acid derivatives coupled aspartic acid (9f: IC50â¯=â¯382⯵m and 10f: IC50â¯=â¯364⯵m, respectively) have the best water solubility and thus presented higher inhibitory activity than that of acarbose (IC50â¯=â¯484⯵m). Unfortunately, all of the derivatives possess lower inhibitory properties of α-glucosidase than those of the parent compounds in the measurement system of DMSO solution, even if the derivatives exhibit better water solubility than that of the parent compounds.
Assuntos
Aminoácidos/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Triterpenos/farmacologia , alfa-Glucosidases/metabolismo , Aminoácidos/química , Relação Dose-Resposta a Droga , Inibidores de Glicosídeo Hidrolases/síntese química , Inibidores de Glicosídeo Hidrolases/química , Humanos , Conformação Molecular , Solubilidade , Relação Estrutura-Atividade , Triterpenos/síntese química , Triterpenos/química , Água/químicaRESUMO
In this study, a simple, rapid and sensitive method combining surface-enhanced Raman spectroscopy and Schiff base reaction was developed for the detection of tryptophan. This method does not require product separation to obtain a significant Raman signal of the derivatized product, and the derivatization reaction can be controlled by experimental parameters such as reaction temperature, time, concentration of derivatization reagent and concentration of sodium nitrite. The characteristic peak of the derivative of tryptophan (1620 cm-1) was selected for quantitative analysis, and the intensity of the characteristic Raman spectrum peak showed a linear relationship with the concentration of tryptophan (10-8-10-4 mol/L) in the range of with a correlation coefficient R2 of 0.9922. This assay combines surface-enhanced Raman spectroscopy and Schiff base reaction, which is characterized by high sensitivity and easy operation, and has good application prospects in the detection of tryptophan in food.
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Nanopartículas Metálicas , Triptofano , Triptofano/química , Prata/química , Análise Espectral Raman/métodos , Bases de Schiff , Nanopartículas Metálicas/químicaRESUMO
A near-infrared fluorescent "turn on" probe DTMI featuring simple skeleton was constructed easily. It undergoes a structure transformation from an A-π-A to a D-π-A framework towards SO32-. Besides, DTMI is capable of distinctive sensing sulfite with a fast response and a significant Stokes shift as well as with high sensitivity, excellent selectivity, long-term stability of fluorescence signals, and good anti-interference ability. The detection limit (LOD) of DTMI for sulfite within the linear concentration range of 0.5-10 µM is 27.39 nM. More importantly, DTMI has been favorably utilized for detecting sulfite in food samples such as red wine and vermicelli. Based on its low biotoxicity, DTMI has been successfully applied in imaging experiments involving HeLa cells, onion inner epidermal cells, and zebrafish. Therefore, the results show that the presented probe possesses potential sensing activity towards sulfite in complex biological system and food samples.
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Corantes Fluorescentes , Sulfitos , Peixe-Zebra , Sulfitos/análise , Sulfitos/química , Humanos , Células HeLa , Corantes Fluorescentes/química , Animais , Cebolas/química , Limite de Detecção , Análise de Alimentos/métodos , Imagem Óptica/métodos , Vinho/análise , Espectrometria de Fluorescência/métodos , Contaminação de Alimentos/análiseRESUMO
A facile, fast and visible sensing platform for ascorbic acid (AA) detection has been developed based on self-assembled hydrangea-like europium metal-organic framework (HL-EuMOF). HL-EuMOF was synthesized through a simple one-step mixing process with Eu3+ and 1, 10-phenanthroline-2, 9-dicarboxylic acid at room temperature, which exhibited excellent properties including strong red fluorescence, long decay lifetime (548.623 µs) and good luminescent stability. Based on the specific redox reaction between Fe3+ and AA, the HL-EuMOF@Fe3+ was fabricated with "turn-off" response for AA, where the resulting Fe2+ displayed effective fluorescence quenching ability toward HL-EuMOF. The sensor demonstrated low detection limit (31.94 nM), rapid response time (30 s) and high selectivity. Integration of smartphone-assisted RGB analysis with HL-EuMOF@Fe3+ permitted convenient and visible quantitative determination of AA level. This approach also presented good detection performances in complex human serum and beverage samples, which could provide a valuable tool for AA detection in biomedical research and food industry.
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Ácido Ascórbico , Európio , Estruturas Metalorgânicas , Oxirredução , Ácido Ascórbico/química , Ácido Ascórbico/análise , Európio/química , Estruturas Metalorgânicas/química , Humanos , Limite de Detecção , Espectrometria de FluorescênciaRESUMO
By coupling thin-film microextraction (TFME) with surface enhanced Raman scattering (SERS), a facile method was developed for the determination of thiram in the complex matrix (orange juice or grape peel). The substrate of TFME was made by self-assembling silver sol on the silicon wafer to form a three-dimensional (3D) silver nanonetwork structure, without adding any template, which was used for TFME and SERS detection, respectively. The substrate exhibits high reproducibility with a relative standard deviation of about 7.32 % in spot and spot SERS intensity. The SERS signal intensity at a shift of 1384 cm-1 and the thiram concentration showed good linearity in the range of 0.01-5 µg/L and the linear correlation coefficient was 0.9912. The detection limit for thiram was found to be 0.01 µg/L. The TFME-SERS method was applied for the determination of thiram in fruit juice and the results were obtained very well. Therefore, this method is expected to play a role in the detection of trace pollutants.
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Análise Espectral Raman , Tiram , Prata , Reprodutibilidade dos Testes , Sucos de Frutas e VegetaisRESUMO
To determine gentamicin residues in animal tissues, a monoclonal antibody (Mab) was produced and a sensitive indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) was developed. At first, gentamicin was conjugated with bovine serum albumin as immunogens which were used to immunize BALB/c mice. Then, an anti-gentamicin Mab was prepared by hybridoma technology. Finally, a sensitive icCLEIA was developed with an 50% inhibition concentration (IC50) of 0.067 ng/mL for gentamicin. The limit of detection of the icCLEIA was 0.002 ng/mL. The cross reactivity of the Mab with structural analogues were<0.01%. The recoveries of gentamicin ranged from 80 to 101% and coefficient of variation was <6.4% in pork and fish samples. Samples were detected by UPLC-MS/MS for evaluating reliability of the icCLEIA. The results suggested that the prepared anti-gentamicin Mab can be used for rapid and convenient immunoassays to detect gentamicin residues in animal tissues.
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Anticorpos Monoclonais , Gentamicinas , Animais , Cromatografia Líquida , Ensaio de Imunoadsorção Enzimática/métodos , Imunoensaio/métodos , Técnicas Imunoenzimáticas , Luminescência , Camundongos , Camundongos Endogâmicos BALB C , Reprodutibilidade dos Testes , Soroalbumina Bovina , Espectrometria de Massas em TandemRESUMO
In the present study, a novel simple and sensitive method was developed for the determination of pectin based on the decarboxylation derivatization reaction and surface-enhanced Raman scattering (SERS) without complicated separation steps. The derivatization reaction can be controlled by the experimental parameters such as reaction time, temperature and the amount of hydrochloric acid. Additionally, the method was used to accurately and reliably detect pectin added in honey or apple, which can be detected at levels as low as 0.5 or 0.01 ppm, respectively. Based on the conventional decarboxylation reaction, a simple and sensitive SERS method was proposed for the detection of pectin, which shows potential for practical application.
Assuntos
Mel , Malus , Descarboxilação , Malus/química , Pectinas , Análise Espectral RamanRESUMO
A novel efficient metal free sensitizer containing asymmetric double donor-π-acceptor chains (DC) was synthesized for dye-sensitized solar cells (DSSCs). Comparing to 3.80%, 4.40% and 4.64% for the DSSCs based on the dyes with single chain (SC1, SC2) and cosensitizers (SC1 + SC2), the overall conversion efficiency reaches 6.06% for DC-sensitized solar cells as a result of its longer electron lifetime and higher incident monochromatic photon-to-current conversion efficiency.
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Prior to the maturation of next-generation energy storage devices, the actual lithium-ion batteries for commercial purposes are still expected to fulfill some critical requirements, among which the high energy density, wide operating temperature range, and related long-term cycling stability are the most challenging issues. Herein a multiple additives strategy is employed to simultaneously optimize the solid electrolyte interphase on the large-area anode and cathode in a 2 Ah artificial graphite (AGr)/LiNi0.5Co0.2Mn0.3O2 (NCM523) pouch cell with high gravimetric (>260 Wh kg-1) and volumetric (>630 Wh L-1) energy density. By introducing a rational mixture of electrolyte additives, a highly sulfurized surface layer and a uniform and thin passivation layer are separately formed on the anode and cathode of the AGr/NCM523 pouch cell, exhibiting high storage stability at 60 °C, much improved discharge capacity at -10 and -20 °C, high anodic stability at high voltage of 4.4 V, and stable cyclic performance with a capacity retention of 85.5% after 500 cycles, significantly outperforming the value of 75.7% after only 200 cycles of the cell without additional additives. These results demonstrate the critical effect of simultaneous optimizations of anode and cathode interphase layers to construct stable high-energy-density lithium-ion pouch cells.
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In the mol-ecule of title compound, C(17)H(18)FN(2)O(4)PS, both the benzene ring with its conjunction C atom and the benzothia-zole ring with its conjunction N atom are close to planar (the maximum deviations are 0.0267 and 0.0427â Å for the benzene and benzothiazole rings, respectively), the dihedral angle between the planes of the benzothia-zole and benzene rings is 119.05â (3)°. The mol-ecular packing is stabilized by inter-molecular N-Hâ¯O, C-Hâ¯N and C-Hâ¯F hydrogen bonding, and by C-Hâ¯π and π-π stacking inter-actions [centroid-centroid distances = 2.99â (2), 2.96â (3), 2.88â (2) and 3.773â (4)â Å].
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Developing high-efficiency hetero-catalysts for transesterification reaction is of great importance in the production of biodiesel from Jatropha curcas L. seed oil (JO). Here, we synthesized a series of sulfated TiO2 by treating with varying H2SO4 concentration (SO4 2-/TiO2) and TiO2 catalysts and applied to the transesterification of JO. Furthermore, these heterostructures were characterized by many characterization methods including XRD, FT-IR, N2-adsorption, SEM, TEM, TG, py-IR and NH3-TPD, and their catalytic performance was investigated under various operating conditions. The results reveal that both the Brønsted and Lewis acid sites are presented in the SO4 2-/TiO2 catalysts, while only Lewis-type sites are observed in the TiO2 catalyst. And the acid intensity, surface area and mesoporous volume of catalysts are improved obviously after treating TiO2 with sulfuric acid. Then the SO4 2-/TiO2 catalysts exhibit much higher catalytic activity than TiO2 catalyst, which is attributed to the larger surface area and mesoporous volume and stronger acidity. Furthermore, the reusability behaviour of 1.5 SO4 2-/TiO2 catalyst in the transesterification of JO was also studied.
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A series of compounds were synthesized by glycosylation of maslinic acid (MA) and corosolic acid (CA) with monosaccharides and disaccharides, and the structures of the derivatives were elucidated by standard spectroscopic methods including (1)H NMR, (13)C NMR and HRMS. The α-glucosidase inhibitory activities of all the novel compounds were evaluated in vitro. The solubility and inhibitory activity of α-glucosidase assays showed that the bis-disaccharide glycosides of triterpene acids possessed higher water solubility and α-glucosidase inhibitory activities than the bis-monosaccharide glycosides. Among these compounds, maslinic acid bis-lactoside (8e, IC50 = 684 µM) and corosolic acid bis-lactoside (9e, IC50 = 428 µM) had the best water solubility, and 9e exhibited a better inhibitory activity than acarbose (IC50 = 478 µM). However, most of glycosylated derivatives possessed lower inhibitory activities than the parent compounds, although their water solubility was enhanced obviously. Moreover, the kinetic inhibition studies indicated that 9e was a non-competitive inhibitor, and structure-activity relationships of the derivatives are also discussed.