Food & Function
·
2022
·Vol. 13
·期 7
·3917-3930
·
IF 6.317
Yan Qiao,Xianfeng Ye,Lingli Zhong,Chengyao Xia,Lei Zhang,Fan Yang,Yongkai Li,Xiaodong Fang,Lei Fu,Yan Huang,Hui Cao,Zhoukun Li,Zhongli Cui
The β-glucan from Saccharomyces cerevisiae is a potent adjuvant that exhibits a broad spectrum of biological activities and health benefits, and different processes have been established to prepare active β-glucan from yeast. However, studies concerning the effect of β-1,6-glucanase enzymolysis on the structure and immunomodulatory activity of yeast β-1,3-glucan are scarce. In this study, we aim to develop a novel enzymatic process for the preparation of immunologically active β-glucan (BYG) from baker's yeast using a β-1,6-glucanase GluM. The β-1,6-glucan in fungal cell wall was specifically hydrolyzed by GluM, and resulted in cell wall decomposition and β-glucan release. Batch production of BYG was realized with 17.8% yield, 85.3% purity and 75.4% recovery rate. Structural characterization indicated that BYG exhibits rod-like structures with natural triplex and nanoparticle-like substructures compared with the commercial Glucan 300. BYG ameliorated inflammation in a DSS-induced mouse model of colitis through inhibiting oxidative stress (NO, MDA and MPO), inflammatory mediators (NLRP3, ASC, caspase-1, iNOS and COX-2), and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, IFN-γ), increasing the expression levels of tight junction proteins (ZO-1, occludin and claudin-1) and modulating the production of gut microbiota-synthesized SCFAs compared to the control. Our results showed that yeast β-1,3-glucan prepared with β-1,6-glucanase exhibits structural integrity that is responsible for its favorable immunomodulatory activity.
研究领域:
微生物学
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免疫学
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生物技术
·
药理学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 3
·1-9
·
IF 5.833
Zhu Jing,Liu Xia,Zheng Jinxue,Jiang Dafeng
A dual channel method is described for the determination of the amyloid-β peptide Aβ(1–40) that is associated with Alzheimer’s disease. The method exploits (a) conformational changes of a G-quadruplex that are triggered by Na + and K + ions and (b) the strong affinity between Aβ(1–40) and Cu 2+ . A G-quadruplex DNA forms an antiparallel structure in the presence of Na + and can catalyze the oxidation of tetramethylbenzidine by H 2 O 2 system in the presence of Cu 2+ to form a visible blue color. If, however, Cu 2+ binds to Aβ(1–40), the blue color is no longer formed. Measuring the absorption decrease at 452 nm, the determination of Aβ(1–40) is realized. If K + is added to the Na + -containing buffer, the antiparallel G-quadruplex DNA is transformed to parallel. This leads to the insertion of protoporphyrin IX (PPIX) into the G-quadruplex and generates enhanced fluorescent signal, with excitation/emission wavelength at 410/630 nm. The G-quadruplex then catalyzes the metalation of PPIX by Cu 2+ , and the fluorescence intensity decreases. In the presence of Aβ(1–40), the formation of Aβ(1–40)-Cu 2+ triggers the recovery of the fluorescence. The Na + /K + -induced tuning of the conformation of the G-quadruplex with the same sequence enables dual (colorimetric and fluorometric) determination of Aβ(1–40), with detection limits of 4.9 pM and 2.3 pM, respectively. The cost is quite low since the developed strategy is label free and enzyme free by using low-cost DNA and Cu 2+ . More importantly, the dual channel determination operation is very simple without any further modification process. Graphical abstract
Tuning the conformation of G-quadruplexes by sodium(I) and potassium(I): application to photometric and fluorometric determination of amyloid β(1–40).
研究领域:
生物化学
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医学诊断
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分子生物学
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DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2020
·Vol. 8
·期 11
·2307-2320
·
IF 5.344
Xiaonan Li,Ziming Zhao,Yihua Yang,Zhaorong Liu,Jinglei Wang,Yalu Xu,Yanzhuo Zhang
In this study, a new type of β-1,3-D-glucan porous microcapsule (GPM)-enveloped and folate conjugated chitosan-functional liposome (FCL), FCL@GPM, was developed for the potential oral co-delivery of chemotherapeutic drugs and quantum dots (QDs) with facilitated drug absorption and antitumor efficacy. In this dual-particulate system, multiple FCLs serve as the cores for effective loading, folate-mediated tumor-targeting, facilitated intracellular accumulation, and pH-responsive controlled release of chemotherapeutic agents, while a GPM acts as the shell for affording macrophage-mediated tumor selectivity. Gefitinib (GEF) was selected as a chemotherapeutic agent, while acid degradable ZnO QDs were selected due to their dual role as an anticancer agent for synergistic chemotherapy and as a fluorescent probe for potential cancer cellular imaging. The GEF and ZnO QD co-loaded FCL@GPMs (GEF/ZnO-FCL@GPMs) exhibited a prolonged release manner with limited release before uptake by intestinal cells. Furthermore, Peyer's patch uptake, macrophage uptake, cytotoxicity, and biodistribution of FCL@GPMs were tested. In addition, GEF and ZnO QD co-loaded FCLs (GEF/ZnO-FCLs) not only have a tumor acidity responsive release property, but also induce a superior cytotoxicity on cancer cells as compared to GEF. Moreover, a 1.75-fold increase in the bioavailability of GEF delivered from GEF/ZnO-FCL@GPMs as compared to its trademarked drug (Iressa®). As a result, GEF/ZnO-FCL@GPMs exerted a superior antitumor efficacy (1.47-fold) as compared to the trademarked drug in mice. Considered together, the developed FCL@GPMs, combining the unique physicochemical and biological benefits of FCLs and GPMs, possess great potential as an efficient delivery system for the co-delivery of chemotherapeutic agents and quantum dots.
研究领域:
药剂学
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肿瘤学
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生物材料学
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DOI
PubMed
高影响因子
ACS Catalysis
·
2019
·
IF 12.221
Hao Cao,Lichao Sun,Ying Huang,Xin Liu,Dong Yang,Tengfei Liu,Xiaojing Jia,Boting Wen,Tianyi Gu,Fengzhong Wang,Fengjiao Xin
Enzymes are usually characterized by their evolutionarily conserved catalytic domains; however, this work presents the incidental gain-of-function of an enzyme in a loop region by natural evolution of its amino acids. A bifunctional acetyl ester–xyloside hydrolase (CLH10) was heterologously expressed, purified, and characterized. The primary sequence of CLH10 contains the fragments of the conserved sequence of esterase and glycosidase, which distribute in a mixed type. The crystal structure revealed that the primary sequence folded into two independent structural regions to undertake both acetyl esterase and β-1,4-xylanase hydrolase functions. CLH10 is capable of cleaving both the β-1,4-xylosidic bond-linked main chain and the ester bond-linked acetylated side chain of xylan, which renders it valuable because it can degrade acetylated xylan within one enzyme. Significantly, the β-1,4-xylanase activity of CLH10 appears to have been fortuitously obtained because of the variable Asp10 and Glu139 located in its loop region, which suggested that the exposed loop region might act as a potential hot-spot for the design and generation of promising enzyme function in both directed evolution and rational protein design.
研究领域:
生物化学
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酶学
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结构生物学
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分子进化
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DOI
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2021
·Vol. 105
·期 12
·5147-5158
·
IF 4.813
Li Qian,Zhu Xiaoman,Xie Yanli,Zhong Yue
Abstract o -Vanillin is a natural product that has been widely applied in the food and pharmaceutical industries. In this study, we determined that o -vanillin can strongly inhibit the growth of Aspergillus flavus mycelia. However, the inhibition mechanism of o -vanillin is still elusive. The ultrastructural morphology of mycelia was injured, and the cell walls were destroyed. The OH functional groups on cell walls were altered, and the content of protein in mycelial cell walls was reduced by o -vanillin. The content of β-1,3-glucan in cell walls was significantly ( P < 0.05) reduced by o -vanillin in a dose-dependent manner, while chitin was not markedly affected. Moreover, o -vanillin led to an increase in the permeability of cell membranes. o -Vanillin also exhibited a promising antifungal effect on contaminated corn kernels. Therefore, o -vanillin inhibited the growth of mycelia by disrupting the integrity of cell walls and cell membranes. This study not only sheds light on the antifungal mechanism of o -vanillin but also indicates that it is a promising agent for the control of A. flavus infection. Key points o -Vanillin has strong inhibitory effects on A. flavus. o -Vanillin destroyed the integrity of cell walls and cell membranes. o -Vanillin could effectively inhibit the growth of A. flavus on corn kernels.
研究领域:
药理学
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微生物学
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食品科学
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DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2025
·
IF 4
Xuemei Lu,Ke Hu,Minggui Ou,Min Li,Xinchun Zhang,Xingyu Zhan,Xun Liao,Ming Li,Rongyu Li
The soft rot disease caused by Fusarium oxysporum leads to a significant reduction in the yield of Dendrobium officinale . However, research into the synergistic inhibitory effect of essential oils in D. officinale is extremely limited. In this study, we systematically investigated the direct and indirect inhibitory activity of carvacrol, citral and their combination against F. oxysporum , and their synergistic inhibitory mechanism. Carvacrol and citral exhibited significant direct and indirect inhibitory activity against F. oxysporum with EC 50 values of 54.37 mg/L and 119.32 mg/L (direct), 18.01 μL/(L•air) and 48.70 μL/(L•air) (indirect). Synergistic analysis revealed that the optimal synergistic toxicity of carvacrol and citral combination (Ca•Ci) against F. oxysporum was 10:1, with co-toxicity coefficient (CTC) of 131.57 and EC 50 value of 44.24 mg/L. Microscopy confirmed that the Ca•Ci led to more significant tip constriction, uneven surfaces and serious rupture of F. oxysporum mycelia than single compound. Transmission electron microscopy (TEM) showed that Ca•Ci also caused significant ultrastructural alterations to F. oxysporum , manifesting as cytoplasmic disorganization and partial organellar disintegration. Moreover, Ca•Ci dramatically increased the sensitivity of F. oxysporum to calcofluor white compared with a single compound. Ca•Ci also considerably upregulated the expression of chitinase-related gene ( FOXG _12882) and β -1,3-glucanase-related gene ( FOXG _10637) in F. oxysporum , resulting in higher chitinase and β -1,3-glucanase activity. However, it should be noted that carvacrol exerted a greater contribution than citral. In conclusion, the combination of carvacrol and citral greatly disrupted the cell wall integrity of F. oxysporum , thereby exhibiting a synergistic effect.
研究领域:
植物病理学
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微生物学
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植物化学
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真菌学
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农业科学
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产品关联
DOI
PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2020
·Vol. 44
·期 1
·67-79
·
IF 2.419
Zhou Hai-Yan,Zhou Jian-Bao,Yi Xiao-Nan,Wang Yan-Mei,Xue Ya-Ping,Chen De-Shui,Cheng Xin-Ping,Li Mian,Wang Hong-Yan,Chen Kai-Qian,Liu Zhi-Qiang,Zheng Yu-Guo
To develop new cellulases for efficient utilization of the lignocellulose, an endoglucanase (CoCel5A) gene from Colletotrichum orchidophilum was synthesized and a recombinant Pichia pastoris GS115/pPIC9K/cocel5A was constructed for secretory expression of CoCel5A. After purification, the protein CoCel5A was biochemically characterized. The endoglucanase CoCel5A exhibited the optimal activity at 55–75 °C and high thermostability (about 85% residual activity) at the temperature of 55 °C after incubation for 3 h. The highest activity of CoCel5A was detected when 100 mM citric acid buffer (pH 4.0–5.0) was used and excellent pH stability (up to 95% residual activity) was observed after incubation in 100 mM citric acid buffer (pH 3.0–6.0) at 4 °C for 24 h. Carboxymethyl cellulose sodium salt ( n = approx. 500) (CMC) and β - d -glucan were the best substrates for CoCel5A among the tested substrates. The kinetic parameters V max , K m , and K cat / K m values against CMC were 290.70 U/mg, 2.65 mg/mL, and 75.67 mL/mg/s, respectively; and 228.31 U/mg, 2.06 mg/mL, and 76.45 mL/mg/s against β - d -glucan, respectively, suggesting that CoCel5A has high affinity and catalytic efficiency. These properties supported the potential application of CoCel5A in biotechnological and environmental fields.
研究领域:
生物化学
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生物技术
·
酶学
·
微生物学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Hao Liu,Min Huang,Shuxin Wei,Xiaowen Wang,Yaqin Zhao,Zhengyang Han,Xianfeng Ye,Zhoukun Li,Yanling Ji,Zhongli Cui,Yan Huang
β-1,3-Galactanases selectively degrade β-1,3-galactan, thus it is an attractive enzyme technique to map high-galactan structure and prepare galactooligosaccharides. In this work, a gene encoding exo -β-1,3-galactanase (PxGal43) was screened form Paenibacillus xylanexedens , consisting of a GH43 domain, a CBM32 domain and α-L-arabinofuranosidase B (AbfB) domain. Using β-1,3-galactan (AG-II-P) as substrate, the recombined enzyme expressed in Escherichia coli BL21 (DE3) exhibited an optimal activity at pH 7.0 and 30 °C. The enzyme was thermostable, retaining >70 % activity after incubating at 50 °C for 2 h. In addition, it showed high tolerance to various metal ions, denaturants and detergents. Substrate specificity indicated that PxGal43 hydrolysis only β-1,3-linked galactosyl oligosaccharides and polysaccharides, releasing galactose as an exo -acting manner. The function of the CBM32 and AbfB domain was revealed by their sequential deletion and suggested that their connection to the catalytic domain was crucial for the oligomerization, catalytic activity, substrate binding and thermal stability of PxGal43. The substrate docking and site-directed mutagenesis proposed that Glu191, Gln244, Asp138 and Glu81 served as the catalytic acid, catalytic base, pKa modulator, and substrate identifier in PxGal43, respectively. These results provide a better understanding and optimization of multi-domain bacterial GH43 β-1,3-galactanase for the degradation of arabinogalactan.
研究领域:
生物化学
·
酶学
·
微生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2016
·Vol. 182
·期 1
·261-275
·
IF 1.606
Jiang Cheng,Song Jinzhu,Cong Hua,Zhang Junzheng,Yang Qian
A novel β-1,3-glucanase gene, designated Ccglu17A , was cloned from the biological control fungus Chaetomium cupreum Ame. Its 1626-bp open reading frame encoded 541 amino acids. The corresponding amino acid sequence showed highest identity (67 %) with a glycoside hydrolase family 17 β-1,3-glucanase from Chaetomium globosum . The recombinant protein Ccglu17A was successfully expressed in Pichia pastoris , and the enzyme was purified to homogeneity with 10.1-fold purification and 47.8 % recovery yield. The protein’s molecular mass was approximately 65 kDa, and its maximum activity appeared at pH 5.0 and temperature 45 °C. Heavy metal ions Fe 2+ , Mn 2+ , Cu 2+ , Co 2+ , Ag + , and Hg 2+ had inhibitory effects on Ccglu17A, but Ba 2+ promoted the enzyme’s activity. Ccglu17A exhibited high substrate specificity, almost exclusively catalyzing β-1,3-glycosidic bond cleavage in various polysaccharoses to liberate glucose. The enzyme had a Km of 2.84 mg/mL and Vmax of 10.7 μmol glucose/min/mg protein for laminarin degradation under optimal conditions. Ccglu17A was an exoglucanase with transglycosylation activity based on its hydrolytic properties. It showed potential antifungal activity with a degradative effect on cell walls and inhibitory action against the germination of pathogenic fungus. In conclusion, Ccglu17A is the first functional exo-1,3-β-glucanase to be identified from C. cupreum and has potential applicability in industry and agriculture.
研究领域:
微生物学
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酶学
·
分子生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2010
·
IF 5.214
Linyan Yang,Li Li,Qin Tu,Li Ren,Yanrong Zhang,Xueqin Wang,Zhiyun Zhang,Wenming Liu,Liangliang Xin,Jinyi Wang
An improved approach for the surface modification of poly(dimethylsiloxane) (PDMS) using carboxymethyl cellulose (CMC), carboxymethyl β-1,3-dextran (CMD), and alginic acid (AA) was investigated. The PDMS substrates were first oxidized in a H2SO4/H2O2 solution to transform the Si−CH3 groups on their surfaces into Si−OH groups. Then methacrylate groups were grafted onto the substrates through a silanization reaction using 3-(trimethoxysilyl)propyl methacrylate. Sequentially, cysteamine was conjugated onto the silanized surfaces by the reaction between the thiol and methacrylate groups under 254 nm UV exposure. Afterward, the amino-terminated PDMS substrates were sequentially reacted with CMC, CMD, and AA in the presence of N-hydroxysuccinimide and 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide, resulting in the grafting of polysaccharides onto PDMS surfaces. The composition and chemical state of the modified surfaces were characterized by X-ray photoelectron spectroscopy (XPS). In addition, the stability and dynamic characteristics of the polysaccharide-grafted PDMS substrates were investigated by XPS and temporal contact angle experiments. A protein adsorption assay using bovine serum albumin (BSA), chicken egg albumin, lysozyme, and RNase-A showed that the introduction of CMD and AA can reduce the adsorption of negatively charged BSA and chicken egg albumin, but increase the adsorption of the positively charged lysozyme and RNase-A. However, CMC-modified PDMS surfaces showed protein-repelling properties, regardless of whether the protein was positively or negatively charged. A cell culture and migration study of glioma C6, MKN-45, MCF-7, and HepG-2 cells revealed that the polysaccharide-modified PDMS greatly improved the cytocompatibility of native PDMS.
研究领域:
材料科学
·
生物材料
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
Biology-Basel
·
2020
·Vol. 9
·期 2
·37
·
IF 3.796
Bo Wu,Qi Wang,Jie Cao,Jun Mei,Jing Xie
Transport in water is the most common method for achieving high survival rates when transporting cultured fish in China; yet, transport success relies on proper water quality and conditions. This research was designed to explore the effects of ascorbic acid and β-1,3-glucan on survival, physiological responses, and flesh quality of farmed tiger grouper (Epinephelus fuscoguttatus) during simulated transport. The transport water temperature for live tiger grouper was 15 °C, which had the highest survival rate, the lowest stress response, and metabolic rate, and this will reduce the susceptibility to diseases. It is stated that β-1,3-glucan influences the changes of cortisol content, heat shock protein 70, IL-1β, and IgM transcription levels during simulated transport. Rather than using ascorbic acid alone (the A-group), β-1,3-glucan (3.2 mg/L) in the presence of ascorbic acid (25 mg/L) can effectively reduce the increase of transport-induced serum cortisol content, heat shock protein 70, and IL-1β, but stimulated IgM. 25 mg/L ascorbic acid and 3.2 mg/L β-1,3-glucan had no obvious effect on the nutritional indexes and flavor of live tiger grouper; however, these can effectively reduce the stress response, improve the innate immune activity, and ensure a higher survival rate.Keywords:Epinephelus fuscoguttatus;simulated transport in water;environment stress;β-1,3-glucan;physiological responses
研究领域:
水产养殖
·
鱼类生理学
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免疫学
·
运输生物学
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产品关联
DOI
PubMed
高影响因子
Catalysis Science & Technology
·
2022
·Vol. 12
·期 6
·1968-1977
·
IF 6.177
Yiwen Yang,Haifeng Qi,Zhanwei Xu,Z. Conrad Zhang
In this work, we identified the prevailing catalytically active species evolved from paired PdCl2–CuCl2 in 1-butyl-3-methylimidazolium chloride ([BMim]Cl) that displayed dramatic synergy between the two metal chlorides in catalyzing the hydrolysis of β-1,4-glycosidic bonds in cellulose. Ultraviolet-visible (UV-vis), far infrared (FIR), and electron paramagnetic resonance (EPR) spectroscopy provide clear evidence for the prominent evolutionary pathways to form the catalytically active species. It is observed that the strong synergy between [PdIICl4]2− and [CuIICl4]2− is initiated by the presence of a saccharide. In the absence of a saccharide, the spectroscopic features of [PdIICl4]2− and [CuIICl4]2− in their paired solution show little change with respect to those of individual metal chloride solutions. The evolution of a highly active heterometallic PdII–Cl–CuI complex from paired PdCl2–CuCl2 in the presence of saccharides is revealed by the transition of the FIR absorption peak of PdII–Cl. Glycolaldehyde as the simplest model compound, which is the same as the reducing end group of saccharides, is applied to determine the possible catalytic species. X-ray absorption spectroscopy (XAS) verifies that [CuIICl4]2− is reduced to [CuICl3]2− by glycolaldehyde. The HCl generated from reduced [CuIICl4]2− is also found to contribute to the hydrolysis of cellobiose, serving as another catalytically active species in paired PdCl2–CuCl2/[BMim]Cl. Paired PdCl2–CuCl2 pretreated with glycolaldehyde exhibits dramatically higher activity than direct use of paired PdCl2–CuCl2 in cellobiose conversion. Strikingly, the PdII–Cl–CuI complex is found to be substantially more active than HCl in the hydrolysis of β-1,4-glycosidic bonds as evidenced by the sustained high cellobiose conversion after neutralization of HCl with NaOH.
研究领域:
催化
·
化学工程
·
材料科学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）