INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Jundan Feng,Huimin Li,Yuzheng Lu,Rong Li,Artur Cavaco-paulo,Jiajia Fu
To enhance the enzymatic digestibility of polyethylene terephthalate (PET), which is highly oriented and crystallized, a polyethylene glycol (PEG) surfactant of varying molecular weights was utilized to improve the stability of mutant cutinase from Humicola insolens (HiC) and to increase the accessibility of the enzyme to the substrate. Leveraging the optimal conditions for HiC hydrolysis of PET, the introduction of 1 % w / v PEG significantly increased the yield of PET hydrolysis products. PEG600 was particularly effective, increasing the yield by 64.58 % compared to using HiC alone. Moreover, the mechanisms by which PEG600 and PEG6000 enhance enzyme digestion were extensively examined using circular dichroism and fluorescence spectroscopy. The results from CD and fluorescence analyses indicated that PEG alters the protein conformation, thereby affecting the catalytic effect of the enzyme. Moreover, PEG improved the affinity between HiC and PET by lowering the surface tension of the solution, substantially enhancing PET hydrolysis. This study suggests that PEG holds considerable promise as an enzyme protector, significantly aiding in the hydrophilic modification and degradation of PET in an environmentally friendly and sustainable manner.
研究领域:
环境科学
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生物技术
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材料科学
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酶学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Guanzhang Liu,Hang Yuan,Yaxin Chen,Lei Mao,Chun Yang,Ruifang Zhang,Guangya Zhang
The proliferation of nano-plastic particles (NPs) poses severe environmental hazards, urgently requiring effective biodegradation methods. Herein, a novel method was developed for degrading nano-PET (polyethylene terephthalate) using immobilized cutinases. Nano-PET particles were prepared using a straightforward method, and biocompatible elastin-like polypeptide-magnetic nanoparticles (ELPs-MNPs) were obtained as magnetic cores via biomimetic mineralization. Using one-pot synthesis with the cost-effective precursor tetraethoxysilane (TEOS), silica-coated magnetically immobilized ELPs-tagged cutinase (ET-C@SiO 2 @MNPs) were produced. ET-C@SiO 2 @MNPs showed rapid magnetic separation within 30 s, simplifying recovery and reuse. ET-C@SiO 2 @MNPs retained 86 % of their initial activity after 11 cycles and exhibited superior hydrolytic capabilities for nano-PET, producing 0.515 mM TPA after 2 h of hydrolysis, which was 96.6 % that of free enzymes. Leveraging ELPs biomimetic mineralization, this approach offers a sustainable and eco-friendly solution for PET-nanoplastic degradation, highlighting the potential of ET-C@SiO 2 @MNPs in effective nanoplastic waste management and contributing to environmental protection and sustainable development.
研究领域:
环境科学
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生物技术
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材料科学
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化学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Catalysis
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2024
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IF 11.3
Yi Zheng,Qingbin Li,Pan Liu,Yingbo Yuan,Longyang Dian,Qian Wang,Quanfeng Liang,Tianyuan Su,Qingsheng Qi
Poly(ethylene terephthalate) (PET) is the most abundant polyester plastic and is causing serious environmental pollution. Rapid biological depolymerization of PET waste at a large scale requires powerful engineered enzymes with excellent performance. Here, we designed a computational strategy to analyze the ligand affinity energy of enzymes to PET chains by molecular docking with the dynamic protein conformations, named affinity analysis based on dynamic docking (ADD). After three rounds of protein engineering assisted by ADD, we drastically enhanced the PET-depolymerizing activity of leaf-branch-compost cutinase (LCC). The best variant LCC-A2 depolymerized >90% of the pretreated, postconsumer PET waste into corresponding monomers within 3.3 h at 78 °C, and over 99% of the products was terminal depolymerization products (terephthalic acid and ethylene glycol), representing the fastest PET depolymerization rate reported to date in the bioreactor under optimal condition. Structural analysis revealed interesting features that improved the ligand affinity and catalytic performance. In conclusion, the proposed strategy and engineered variants represent a substantial advancement of the biological circular economy for PET.
研究领域:
环境科学
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生物技术
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材料科学
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蛋白质工程
✓ 已验证
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2019
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IF 4.784
Xiaojie Duan,Zhengqiang Jiang,Yu Liu,Qiaojuan Yan,Man Xiang,Shaoqing Yang
A codon-optimized cutinase gene ( TtCutopt ) from Thielavia terrestris was over-expressed in Pichia pastoris . An extracellular activity reached 10,200 U/mL using high cell density fermentation. The optimal pH and temperature of TtCutopt were 7.0 and 50 °C, respectively. It displayed high stability over a wide range of pH from 3.0 to 11.0 and up to 85 °C. Among tested p -nitrophenyl esters and triglycerides , TtCutopt showed the highest activity towards p -nitrophenyl butyrate and tributyrin, with specificity activity of 2322.4 U/mg and 1152.5 U/mg, respectively. It was extremely stable in organic solvents and surfactants. TtCutopt efficiently catalyzed the synthesis of butyl butyrate , hexyl butyrate, butyl hexanoate and hexyl hexanoate with esterification efficiency of >95%. Furthermore, it catalyzed the degradation of >90% of dimethyl phthalate, diethyl phthalate, dipropyl phthalate and dibutyl phthalate to release their corresponding monoalkyl phthalates within 24 h. Thus, high yield, high stability, and esterification efficiency of TtCutopt make it an attractive candidate for ester biosynthesis and biodegradation.
研究领域:
生物技术
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生物化学
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酶学
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微生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENVIRONMENT INTERNATIONAL
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2024
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IF 10.3
Wenchao Zhan,Chenxu Wang,Xiyu Yang,Hongtao Li,Sicheng Xiong,Xiaoyang Li
A pH-responsive colorimetric method based on dual-enzyme catalysis for rapid and facile detection and quantification of nanoPET at environment-dependent concentration is proposed. The nanoPET was hydrolyzed by the synergistic catalysis of cutinase and lipase to terephthalic acid which can be sensitive detected using bromocresol purple as the indicator. The color changed from purple to bright yellow as the nanoPET detection concentration increased from 0 mg/mL to 2 mg/mL which can be detected by UV–Vis. This naked-eye method has a high sensitivity for nanoPET detection with the visual detection cutoff of 31.00 μg/mL, and has a good linearity in the range of 0 ∼ 1 mg/mL with LOD of 22.84 μg/mL. The reliability of this method is verified in the detection of nanoPET in lake water and beer samples, with an average recovery of 87.1 %. The as-developed dual-enzyme colorimetric chemosensor holds promising potential as a robust and effective platform for the sensitive detection of nanoPET.
研究领域:
环境科学
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分析化学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
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2024
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IF 3.7
Qingjun Jia,Zixuan Zhang,Lankai Su,Shu Bai,Di Cai,Changjing Chen,Linling Yu,Yan Sun
Enzymatic degradation of poly(ethylene terephthalate) (PET) into its corresponding monomers provides an attractive route for green plastic recycling and environmental remediation applications. However, the binding of PET hydrolases toward the PET surface is a rate-limiting step in PET degradation. Hence, we selected three anchor peptides and four linker peptides to fuse the previously engineered leaf-branch compost cutinase for facilitating enzyme binding and improving degradation of real post-consumer PET wastes of Coca Cola and C′eastbon PET bottles, with using the amorphous PET as a control. The systematical characterization of the seven cutinases revealed that the anchor peptide with moderate hydrophobicity plus chaperone activity and the flexible linker peptide with small hydrophobicity but long length were more suitable for each domain to serve its own role, affording the facilitated PET binding affinities and enhanced PET degradation performance. Thus, the optimal fusion cutinase possessed up to 3.3 times higher catalytic efficiency ( k cat / K m ) and 76 % increased long-term degradation performance than the parental cutinase, respectively. Further, complete degradation of C′eastbon PET wastes and 77 % degradation of Coca Cola PET wastes were achieved by the optimal cutinase of 33.3 nmol·g −1 PET in 96 h, proving its superiority against the real post-consumer PET wastes.
研究领域:
环境科学
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生物技术
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材料科学
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化学工程
✓ 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）