CHINESE JOURNAL OF CHEMICAL ENGINEERING
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2023
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IF 3.8
Renwen Tian,Yan Sun
Extensive use of polyethylene terephthalate (PET) has brought about global environmental problems. A recently reported PET hydrolase (PETase) discovered from Ideonella sakaiensis showed high potential for degrading PET at moderate temperatures, but its activity and stability need further improvement for practical applications. Herein, we proposed to use α-synuclein (αS) as a fusion chaperone and created six PETase-αS fusion enzymes with linkers of different types and lengths. All the fusion enzymes exhibited improved enzymatic performance, presenting 1.5 to 2.6-fold higher activity towards bis-2(hydroxyethyl) terephthalate than PETase, as well as significantly increased stabilities. Fluorescence spectroscopy indicated that the chaperone fusion tightened the overall conformation and resulted in the opening of the substrate binding pocket, which led to the improved thermal stability and catalytic activity of the fusion enzymes. Remarkably, one of the fusion proteins, PETase-[(GS)(EK)] 10 -αS, showed 3.2 to 5.1 times higher PET degradation capability than PETase. The significantly boosted PET degradation performance was not only attributed to the enhanced enzymatic activity and stability, but also possibly due to the binding affinity of the fused αS domain for PET. These findings demonstrated that αS was an effective fusion chaperone for significantly enhancing the enzymatic performance of PETase.
研究领域:
酶学
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生物技术
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环境科学
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蛋白质工程
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DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
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2022
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IF 3.705
Jun Cao,Rong Wu,Feng Zhu,Qihui Dong,Erzheng Su
With the development of green chemistry, the demand for environmentally friendly and biocompatible solvents for non-aqueous enzymatic catalysis is increasingly urgent. Deep eutectic solvents (DESs) are viewed as the most promising alternatives to traditional organic solvents in non-aqueous biocatalysis . To expand the types of DESs and provide guidance for DESs design for non-aqueous enzymatic catalysis, the enzyme performance in aqueous buffer after incubation in nearly anhydrous DESs was associated with the properties and component structures of DESs. Almond β- Glucosidase (β-GC) and Candida antarctica lipase B (CALB) were selected as model enzymes . Physico-chemical properties of DESs (as inferred by their solvatochromic parameters) were applied to explore the influences of DESs properties on enzyme activity . For DESs with the same HBD , the biocompability of DESs and thermal stability of enzymes in DESs were negatively associated with the polarity and hydrogen bond acidity of DESs, and were positively associated with hydrogen bond basicity of DESs. Whereas an opposite trend was observed in DESs with the same HBA. Analyzing from the DESs components, the biocompatibility of hydrophobic DESs for enzyme was much lower than that of hydrophilic DESs. Generally, the amount of hydroxyl group and the length of carbon chain represent advantageous ingredients for maintaining natural structure of enzyme molecule. The presence of carboxyl group in hydrophilic DESs and carbon-carbon double bond may impair enzyme structure and activity. This work is hoped to be helpful in expanding the applications of DESs in non-aqueous biocatalysis.
研究领域:
绿色化学
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生物催化
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环境科学
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生物化学
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DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
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2020
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IF 3.503
Shi-Yu Zhang,Ze-Wang Guo,Xiao-Ling Wu,Xiao-Yang Ou,Min-Hua Zong,Wen-Yong Lou
Cold-adapted pullulanase with high catalytic activity and stability is of special interest for its wide application in cold starch hydrolysis, but few pullulanases displaying excellent characteristics at ambient temperature and acidic pH have hitherto been reported. Here, a novel pullulanase from Bacillus methanolicus PB1 was successfully expressed in Escherichia coli BL21 (DE3) and determined to be a cold-adapted type I pullulanase (PulPB1) with maximum activity at 50 °C and pH 5.5. The recombinant PulPB1 showed great stability, its half-life at 50 °C was 137 h. PulPB1 can efficiently hydrolyze pullulan and amylopectin, with activities of 292 and 184 U/mg at 50 °C and pH 5.5, respectively. Moreover, the N-terminal domain of PulPB1 was found to significantly affect the enzymatic performance. Following truncation of the N-terminal domain, the activity towards pullulan decreased markedly from 292 to 141 U/mg and the half-life at 50 °C decreased from 137 to 10 h. Compared to the hydrolysis system with amyloglucosidase alone, the catalytic efficiency showed a 2.4-fold increase on combining PulPB1 with amyloglucosidase for amylopectin hydrolysis at 40 °C. This demonstrates that PulPB1 is promising for development as a superior candidate for cold amylopectin hydrolysis.
研究领域:
生物化学
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微生物学
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酶学
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生物技术
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DOI
PubMed
高影响因子
RSC Advances
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2022
·Vol. 12
·期 6
·3454-3464
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IF 4.036
Weijing Kong,Qiaolin Du,Yinan Qu,Changyu Shao,Chaoqun Chen,Jian Sun,Caiyun Mao,Ruikang Tang,Xinhua Gu
It is currently known that crosslinking agents can effectively improve the mechanical properties of dentin by crosslinking type I collagen. However, few scholars have focused on the influence of crosslinking agents on the collagen-mineral interface after crosslinking. Analysis of the Fourier transform infrared spectroscopy (FTIR) results showed that hydrogen bonding occurs between the tannic acid (TA) molecule and the collagen. The crosslinking degree of TA to collagen reached a maximum 41.28 ± 1.52. This study used TA crosslinked collagen fibers to successfully induce dentin biomineralization, and the complete remineralization was achieved within 4 days. The crosslinking effect of TA can improve the mechanical properties and anti-enzyme properties of dentin. The elastic modulus (mean and standard deviation) and hardness values of the remineralized dentin pretreated with TA reached 19.1 ± 1.12 GPa and 0.68 ± 0.06 GPa, respectively, which were close to those of healthy dentin measurements, but significantly higher than those of dentin without crosslinking (8.91 ± 1.82 GPa and 0.16 ± 0.01 GPa). The interface energy between the surface of collagen fibers and minerals decreased from 10.59 mJ m−2 to 4.19 mJ m−2 with the influence of TA. The current work reveals the importance of tannic acid crosslinking for dentin remineralization while providing profound insights into the interfacial control of biomolecules in collagen mineralization.
研究领域:
牙科材料
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生物材料
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生物矿化
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胶原科学
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材料科学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2023
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IF 8.2
Yu Chen,Danqi Li,Yaping Zhong,Zhentan Lu,Dong Wang
Developing a hydrogel dressing with excellent antibacterial efficacy for accelerating wound healing is high desirable in clinical applications. In this work, NIR regulated metal-organic framework composite hydrogel dressing was constructed for enhanced antibacterial efficacy and accelerated wound healing via the compounding between hydrogel and UCNPs@ZrMOF-Pt nanoparticles. The visible light emitted from upconvertion nanoparticles (UCNPs) activated porphyrin based metal-organic framework (MOF) in composite hydrogel to generate 1 O 2 for photodynamic antibacterial therapy under NIR laser irradiation. Moreover, the UCNPs@ZrMOF-Pt in composite hydrogel with catalase-like performance could effectively convert the high concentration H 2 O 2 in wound to abundant O 2 , which relieved the hypoxic in infected wound. Thus, the photodynamic antibacterial efficacy was remarkably enhanced, leading to accelerate the wound healing. This work presented a novel strategy for high efficient antibacterial therapy and accelerated wound healing.
研究领域:
生物医学工程
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材料科学
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纳米医学
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产品关联
DOI
PubMed
高影响因子
BIOMACROMOLECULES
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2018
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IF 5.738
Xue-Jian Zhang,Xiao-Wei Wang,Jia-Xing Sun,Chao Su,Shuguang Yang,Wen-Bin Zhang
Protein immobilization is critical to utilize their unique functions in diverse applications. Herein, we report that orthogonal peptide–protein chemistry enabled multilayer construction can facilitate the incorporation of various folded structural domains, including calmodulin in different states, affibody, and dihydrofolate reductase (DHFR). An extended conformation is found to be the most advantageous for steady film growth. The resulting protein thin films exhibit sensitive and selective responsive behaviors to biosignals, such as Ca2+, trifluoperazine, and nicotinamide adenine dinucleotide phosphate (NADPH), and fully maintain the catalytic activity of DHFR. The approach is applicable to different substrates such as hydrophobic gold and hydrophilic silica microparticles. The DHFR enzyme can be immobilized onto silica microparticles with tunable amounts. The multilayer setup exhibits a synergistic enhancement of DHFR activity with increasing numbers of bilayers and also makes the embedded DHFR more resilient to lyophilization. Therefore, this is a convenient and versatile method for protein immobilization with potential benefits of synergistic enhancement in enzyme performance and resilience.
研究领域:
生物化学
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材料科学
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生物技术
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酶学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
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IF 8.3
Zhi Qu,Yan Sun
The enormous accumulation of poly(ethylene terephthalate) (PET) waste has posed a serious threat to the environment and human health, and biodegradation with PET hydrolase (PETase) can be a possible solution. Herein, we propose site-specifically modifying PETase with amphiphilic polymers to improve the enzyme performance at ambient temperature. For this purpose, we devise a computer-aided strategy to prioritize the conjugation site, and polyethylene glycol (PEG) preparations of 0.55 to 10 kDa are site-specifically conjugated to PETase. The most active conjugate PETase–PEG 5k (PETase-5K) shows an increase of melting temperature (3.88 °C) and significantly improves PET degradation performance (3.5- and 3.1-fold increases at 30 and 40 °C, respectively). Experimental investigation and molecular dynamics simulations reveal that the site-specific PEGylation increases the hydrophobic solvent-accessible surface area and the binding capability to the PET surface, thickens the hydration layer, increases the intramolecular hydrogen bonding, reduces the interactions between water and the conjugated enzyme surface, and rigidifies the enzyme structure via hydrogen bonding and hydrophobic interactions between the polymer and the enzyme, thus leading to improved enzymatic performance of PETase-5K. We further validate the versatility of the site-specific PEGylation in one of the most evolved variants of PETase, FAST-PETase, by 1.8-fold improvement in PET degradation at 30 °C. The presented computer-aided site-specific conjugation strategy has opened a new avenue to enhancing PETase performance at ambient temperature, and the contribution of PEGylation to PETase unraveled in this work laid a foundation for the rational engineering of PET hydrolases.
研究领域:
环境科学
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生物化学
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生物技术
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分子生物学
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材料科学
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产品关联
DOI
PubMed
高影响因子
Small
·
2025
·
IF 12.1
Qiao Chen,Xu Yang,Yao Yu,Xinmei Duan,Rongrong Ni,Guojing Song,Li Zhu,Yuan Zhong,Kai Qu,Xian Qin,Kun Zhang,Yang Luo,Wei Wu
Theranostic applications in atherosclerosis plaque microenvironment-triggered nanoplatforms are significantly compromised by the complex synthesis procedure, non-specific distribution, and limited therapeutic function. Therefore, development of a facile and feasible method to construct a pathology-based stimuli-responsive nanoplatform with satisfactory theranostic performance remains a demanding and highly anticipated goal. Herein, a novel class of multifunctional supra-carbon dots (CDs), denoted as MM@Ce-CDs NPs, by a simple nanoassembly and a subsequent coating with macrophage membrane (MM), is developed for the targeted reactive oxygen species-trigged theranostic and positive regulation of the pathological plaque microenvironment in AS. The harvested MM@Ce-CDs NPs exhibit activatable fluorescence properties, photoacoustic characteristics, and cascade enzyme performances, which can be effectively activated under ROS stimulation in the plaque pathological microenvironment, enabling precise control over theranostic functions, while markedly enhancing diagnostic accuracy and therapeutic efficacy for AS management. Besides, MM@Ce-CDs NPs can effectively manipulate the plaque microenvironment by reducing ROS levels and inflammation, alleviating M1 macrophage infiltration, and inhibiting foam cell formation, all together suppressing the pathological plaque development through the synergistic mechanisms. In addition, MM@Ce-CDs NPs inherit the biomimetic biological functions from MM, facilitating a highly specific target delivery to AS.
研究领域:
纳米技术
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生物医学工程
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心血管疾病
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材料科学
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诊疗一体化
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产品关联
DOI
PubMed
高影响因子
ACS Applied Polymer Materials
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2022
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IF 4.855
Hao-Chen Li,Xiao-Min Sun,Yong-Rui Huang,Yue-Hai Peng,Jia Liu,Li Ren
Collagen can self-assemble to form fibers with a unique structure and retain a reticular fibrous structure after membrane formation. However, collagen membranes without further modification are easy to tear. In this study, two crosslinkers (linear di-alkynyl poly(ethylene glycol) (DA-PEG) and branched tetra-alkynyl poly(ethylene glycol) (4arm-DA-PEG)) were synthesized, and exogenous crosslinking between collagen fiber bundles through the amino–yne click reaction was studied. Fourier-transform infrared spectroscopy (FT-IR) was performed to confirm the successful synthesis of crosslinkers and the construction of the crosslinked network. Scanning electron microscopy (SEM) showed that the crosslinked collagen membranes (Col/DA-PEG; Col/4arm-DA-PEG) had dense fibrous structures, which was expected to provide a structural foundation for surgical suturing. The denser structures and higher crosslinking degree of Col/4arm-DA-PEG provided good mechanical, optical, and antienzymatic properties. Col/4arm-DA-PEG could withstand 4.6 N/mm suture strength, which enabled it for suturing. Compared with glutaraldehyde-crosslinked membranes, Col/DA-PEG and Col/4arm-DA-PEG showed low cytotoxicity and better biocompatibility, as observed by CCK-8 assay and confocal laser scanning microscopy. These results indicate that alkynyl-terminated PEG systems can be used as ideal collagen crosslinkers to enhance the suture tension of collagen-based materials, especially in corneal repair materials.
研究领域:
材料科学
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生物材料
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组织工程
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眼科学
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化学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）