CARBOHYDRATE POLYMERS
·
2023
·
IF 11.2
Yazhen Wu,Xiaoxiao Li,Zhengyu Jin,Birte Svensson,Yuxiang Bai
How to improve the solubility of linear dextrins (LD) and retain their characteristic helix amphiphilic cavities with flexible embedding capability, is a question worth exploring without adding new chemical groups. The strategy presented in this study is to attach a highly flexible (α-1 → 6) glucochain at the reducing end of LD by preparing a new type of dextrin, referred to as single-arm linear dextrin (SLD). In the actual synthesis, an (α-1 → 6) linked oligosaccharide of DP ¯ 10.7 (PDI = 1.28) was formed by extension of glucose units onto sucrose (2 M) by using L940W mutant of the glucansucrase GTF180-ΔN firstly. Next using γ-CD as glucosylation donor γ-CGTase extended this (α-1 → 6) glucochain with (α-1 → 4) bonds. SLD is a chimeric glucosaccharide comprising an (α-1 → 4) linked part ( DP ¯ 10.5) attached to the nonreducing end of an (α-1 → 6) glucochain as verified by enzyme fingerprinting and 1 H NMR. Furthermore, SLD was validated to show greatly improved solubility and dispersibility of resveratrol in water, as indicated by a 3.12-fold enhancement over the solubility in the presence of 0.014 M SLD. This study provided a new strategy for solving the solubility problem of LD and opens possibilities for new design of the fine structure of starch-like materials.
研究领域:
生物化学
·
食品科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Huimin Tian,Yachun Gu,Zili Lv,Liang Wang
Yoghurt fermented by Leuconostoc mesenteroides XR1 from Kefir grains was found to produce a unique silk drawing phenomenon. This property was found to be associated with the exopolysaccharides (EPS), X-EPS, produced by strain XR1. In order to better understand the mechanism that produced this phenomenon, the X-EPS was extracted, purified and characterized. The molecular weight and monosaccharide composition were determined by size exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS) and ion chromatography (IC) analysis, respectively. The results showed that its molecular weight was 4.183 × 10 6 g/mol and its monosaccharide composition was glucose, and glucuronic acid , with the contents of 567.6148 and 0.2096 μg/mg, respectively. FT-IR and NMR analyses showed that X-EPS was an α-pyranose polysaccharide and was composed of 92.22 % α-(1 → 6) linked d -glucopyranose units and 7.77 % α-(1 → 3) branching. Furthermore, it showed a chain-like microstructure with branches in atomic force microscopy (AFM) and scanning electron microscopy (SEM) experiments. These results suggested that the unique structure of X-EPS, gave the yoghurt a strong viscosity and cohesiveness, which resulted in the silk drawing phenomenon. This work suggested that X-EPS holds the potential for food and industrial applications.
研究领域:
食品科学
·
微生物学
·
聚合物化学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2021
·
IF 7.514
Chenlu Zhu,Zebin Liu,Lishi Ren,Siming Jiao,Xuebing Zhang,Qiukuan Wang,Zhimin Li,Yuguang Du,Jian-Jun Li
Endo -fucoidanases are important in structural analysis of fucoidans and preparation of fuco-oligosaccharides. However their enzymological properties and analysis of degradation products are scarcely investigated. Truncated endo - α (1 → 3)-fucoidanase Fda1 (tFda1B from Alteromonas sp. was overexpressed and characterized, showing highest activity at pH 7.0, 35 °C, and 1.0 M NaCl. Its K m and k cat were 3.88 ± 0.81 mg/mL and 0.82 ± 0.17 min −1 . Fe 3+ and Mn 2+ enhanced activity by 100% and 19.5% respectively. Co 2+ and Cu 2+ completely inactivated tFda1B, whereas Ni 2+ , Mg 2+ , Zn 2+ , Pb 2+ , Ca 2+ , Ba 2+ and Li + decreased activity by 58.8%, 56.0%, 50.6%, 47.7%, 28.9%, 15.6% and 37.5%, respectively. Catalytic residues were identified through structure and sequence alignment, and confirmed by mutagenesis. Degradation products of Kjellmaniella crassifolia fucoidan by tFda1B were characterized by LC-ESI-MS/MS, confirming tFda1B belongs to endo -(1 → 3)-fucoidanases, and backbone of K. crassifolia fucoidan is 1 → 3 fucoside linkage. This endo - α (1 → 3)-fucoidanase would be useful for elucidating fucoidan structures, and be used as a food enzyme.
研究领域:
生物化学
·
酶学
·
海洋生物技术
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Yu Zhang,Niuniu Wu,Jingyi Wang,Zehong Chen,Zhijing Wu,Mengzi Song,Ziming Zheng,Kaiping Wang
Grifola frondosa polysaccharide (GFP) is mainly composed of α-1,4 glycosidic bonds and possesses multiple pharmacological activities. However, the absence of pharmacokinetic studies has limited its further development and utilization. Herein, GFP was labeled with 5-DTAF (FGFP) and cyanine 5.5 amine (GFP-Cy5.5) to investigate its gastrointestinal metabolism characteristics and mechanism. Significant distributions of the polysaccharide in the liver and kidneys were observed by near infrared imaging. To investigate the specific distribution form of the polysaccharide, in vitro digestion models were constructed and revealed that FGFP was degraded in saliva and rat small intestine extract. The metabolites were detected in the stomach and small intestine, followed by further degradation in the distal intestine in the in vivo experiment. Subsequent investigations showed that α-amylase was involved in the gastrointestinal degradation of GFP, and its metabolite finally entered the kidneys, where it was excreted directly with urine.
研究领域:
食品科学
·
生物化学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2024
·
IF 4.3
Zhe-Ying Hu,Huan-Yu Sui,Qi-Feng Zhong,Lu Hu,Jie-Hua Shi,Shao-Liang Jiang,Liang Han
Dacomitinib is a highly selective second-generation tyrosine kinase inhibitor that can irreversibly bind to tyrosine kinase and is mainly used in the treatment of lung cancer. The binding characteristics of dacomitinib with human α 1-acid glycoprotein (HAG) were analyzed by multispectral and computational simulation techniques. The fluorescence spectra showed that dacomitinib can quench the fluorescence of HAG by forming the HAG-dacomitinib complex with a molar ratio of 1:1 (static quenching). At the temperature similar to that of the human body, the affinity of dacomitinib to HAG (8.95 × 10 6 M −1 ) was much greater than that to BSA (3.39 × 10 4 M −1 ), indicating that dacomitinib will give priority to binding onto HAG. Thermodynamics parameters analysis and driving force competition experiments showed that hydrogen bonding and hydrophobic forces were the major sources for keeping the complex of HAG-dacomitinib stable. The experimental outcomes also showed that the binding of dacomitinib can lead to the loosening of the skeleton structure of HAG, which led to a slight change in the secondary structure , and also reduces the hydrophobicity of the microenvironment of Trp and Tyr residues. The binding sites of dacomitinib on HAG and the contribution of key amino acid residues to the binding reaction were determined by molecular docking and molecular dynamics (MD) simulation. In addition, it was found that there was a synergistic effect between dacomitinib and Mg 2+ and Co 2+ ions. Mg 2+ and Co 2+ could increase the K b of dacomitinib to HAG and prolong the half-life of dacomitinib.
研究领域:
药理学
·
生物化学
·
计算生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2024
·
IF 4.3
Huan-Yu Sui,Dong Chen,Jia-Ping Huang,Zhe-Ying Hu,Lu Hu,Jie-Hua shi,Shao-Liang Jiang
Approval in 2019 was granted for the highly selective, targeted agent lorlatinib, which primary target is ROS1 and ALK. The purpose of this work was to examine the binding mechanism between lorlatinib (LOR) and HAG employing multispectral and molecular modeling techniques. Fluorescence data demonstrated that LOR quenched HAG fluorescence as a static quenching, interecalated into the hydrophobic cavity of HAG with a moderate affinity. Thermodynamic and competitive experiments pointed out that LOR bound with HAG primarily through hydrogen bonding, hydrophobic, and van der Waals forces. Circular dichroism, three-dimensional and synchronous fluorescence spectroscopic studies noted that the secondary structure of HAG and microenvironments around tyrosine (Tyr) and tryptophan (Trp) residues were altered due to binding with LOR. The contribution of each energy involved in binding process of LOR and HAG has been analyzed by molecular simulation techniques. Besides, the environmental conditions with metal ions have also been studied. The present study is expected to provide a theoretical basis for further studying the metabolism of LOR in vivo, which may help to gain a deeper understanding of the general pharmacological activity of the drug.
研究领域:
药理学
·
生物化学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Foods
·
2022
·Vol. 11
·期 10
·1442
·
IF 5.561
Yamei Wang,Yixiong Tian,Xiaofeng Ban,Caiming Li,Yan Hong,Li Cheng,Zhengbiao Gu,Zhaofeng Li
Amylo-α-1,6-glucosidase (EC 3.2.1.33, AMY) exhibits hydrolytic activity towards α-1,6-glycosidic bonds of branched substrates. The debranching products of maltodextrin, waxy corn starch and cassava starch treated with AMY, pullulanase (EC 3.2.1.41, PUL) and isoamylase (EC 3.2.1.68, ISO), were investigated and their differences in substrate selectivity and debranching efficiency were compared. AMY had a preference for the branched structure with medium-length chains, and the optimal debranching length was DP 13–24. Its optimum debranching length was shorter than ISO (DP 25–36). In addition, the debranching rate of maltodextrin treated by AMY for 6 h was 80%, which was 20% higher than that of ISO. AMY could decompose most of the polymerized amylopectin in maltodextrin into short amylose and oligosaccharides, while it could only decompose the polymerized amylopectin in starch into branched glucan chains and long amylose. Furthermore, the successive use of AMY and β-amylase increased the hydrolysis rate of maltodextrin from 68% to 86%. Therefore, AMY with high substrate selectivity and a high catalytic capacity could be used synergistically with other enzyme preparations to improve substrate utilization and reduce reaction time. Importantly, the development of a novel AMY provides an effective choice to meet different production requirements.Keywords:amylo-α-1,6-glucosidase;substrate selectivity;pullulanase;isoamylase;debranching efficiency
研究领域:
生物化学
·
酶学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
STAR Protocols
·
2024
·
IF 1.3
Yuanyan Wei,Shuting Geng,Qihang Chen,Jiayue Chen,Zhijun Fan,Yi Li,Yu Si,Yuerong Yang,Ziwei Liang,Jianhai Jiang
Summary CD133 is widely used as a marker to isolate cancer stem cells (CSCs). However, the structural ambiguity of N-glycan of CD133 limits its application in the isolation of CSCs. Here, we present a protocol to sort CSCs from tumor samples by combining CD133 with α-1,2-high-mannose type glycan chains. We describe steps for purifying and biotinylating cyanovirin-N (CVN) proteins and sorting tumor cells and CD133 + α-1,2-Man + (α-1,2-linked mannose + ) cells to provide more specific markers for CSCs. For complete details on the use and execution of this protocol, please refer to Wei et al. 1
研究领域:
肿瘤学
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Shao-Liang Jiang,Wan-Jun Wang,Zhe-Ying Hu,Rong-Juan Zhang,Jie-Hua Shi
The primary coverage of this paper is to investigate the molecular interaction of JAK2 inhibitor, fedratinib (FED) with BSA/HAG proteins through multispectral approaches and molecular docking as well as MD calculation. Arrival at a conclusion, the endogenous fluorescence of BSA/HAG protein was quenched separately in the form of static and mixed quenching. The FED-BSA and FED-HAG complexes with the stoichiometric ratio of 1:1 were formed in the interaction process. And, The K b values of these complexes were of 10 4 –10 5 M −1 and 10 5 –10 6 M −1 , respectively, representing that the FED-HAG complex exhibited a comparatively high affinity compared to the FED-BSA complex. It is confimed that FED inserted into the interface area between subdomain IIA and IIB of BSA (marked as site II') and the bucket-shaped hydrophobic cavity of HAG, respectively, resulting in the slight alteration in the secondary structure of BSA/HAG and the micro-environment round Tyr and Trp residues. The expetimental results also confirmed that van der Waals forces (VDW), hydrogen bonds and hydrophobic interaction played a dominant role in the formation of two stable complexes. The above experimental results were supplemented and verified through molecular docking and MD simulation. Meanwhile, the effects of common ions on affinity were explored. This study could shine a light on evaluating the pharmacological properties of the JAK inhibitor FED, understanding the distribution and operation of the drug in the body, and leading to the development of the creation of novel medication devise.
研究领域:
药理学
·
生物化学
·
计算生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2019
·
IF 6.044
Laifeng Lu,Lifeng Ji,Ruixi Shi,Shuhua Li,Xi Zhang,Qingbin Guo,Changlu Wang,Liping Qiao
Alpha-1,3-glucan is often synthesized on the surface of pathogenic filamentous fungi cell walls to block pathogen-associated molecular patterns (PAMPs) generation by host plant enzymes and the subsequent immune system response of the plant. Here, Botrytis cinerea susceptibility was assessed in tomato fruit to determine whether the fruit could recognize this camouflage and mount an immune response to it. The results showed that local mechanical wounds treated with dextran and laminarin, except amylopectin, could locally and then systemically activate disease resistance against B. cinerea infection in tomato fruit. Dextran treatment effectively elicited fruit callose deposition and phenylpropanoid and flavonoid biosynthesis to a greater extent than α-glucanase activity relative to the mock group surface wounds. Enzymatic hydrolysis of this polysaccharide provided some help in improving host disease resistance. Taken together, these results demonstrate that tomato fruit can perceive α-1,3-glucan as a kind of PAMPs but have limited ability to degrade it.
研究领域:
植物病理学
·
植物生理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Jicheng Niu,Haixiao Shi,Huigang Wei,Bin Gao,Jasmine Xinze Li,Feng Xu,Xiaoguang Li,Fei Li
Developing miniaturized and rapid protein analytical platforms is urgently needed for on-site protein analysis, which is important for disease diagnosis and monitoring. Liquid marbles (LMs), a kind of particle-coated droplets, as ideal microreactors have been used in various fields. However, their application as analytical platforms is limited due to the difficulty of pretreating complex samples in simple LMs. Herein, inspired by the microfluidic chip, we propose a strategy through fabricating fluid channels using deformable LM, termed liquid plasticine (LP), to achieve sample pretreatment function. Through combining isoelectric focusing (IEF) with an LP channel, an LP–IEF platform with simultaneous protein separation and concentration functions is realized. The pretreatment capability of the LP–IEF system for proteins in physiological samples is proven using standard proteins and human serum with the results of a clear separation, 10-fold concentration, and a resolution of 0.03 pH. Through cutting the LP after IEF to LMs and transiting isolated LMs containing target proteins for further downstream colorimetric and mass spectrometry measurements, the quantitative analysis of clinical microalbuminuria and identification of α-1-microglobulin/bikunin precursor in clinical diabetic urine samples are achieved. This work proposes a strategy to develop LMs/LPs as a multifunctional integrated analytical platform and the miniaturized LP–IEF device as a rapid protein analytical platform.
研究领域:
分析化学
·
生物技术
·
生物化学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）