CHEMICAL ENGINEERING SCIENCE
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2024
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IF 4.1
Shaobin Zhang,Junjun Liu,Xuan Wang,Dezhi Liu,Qikai Tian,Haixia Shen,Jing Zhang,Dongpo Song,Weiliang Dong,Ziyi Yu
While enzymatic degradation has proven to be an effective way of remediating nanoplastic-related pollution, the practical deployment of this technique is hampered by concerns over enzyme stability, costs, and the low concentration of nanoplastics in natural settings. In this study, we unveil a facile microreactor-enhanced method that improves the effective remediation capacity of cutinase for nanoplastic-polluted water systems. Our approach leverages the self-assembly capabilities of block copolymers within microfluidic droplets to fabricate Janus microspheres with distinct dual-porosity features, which have an enzyme loading capacity of up to 103.8 mg cutinase/g microsphere. The cutinase-anchored microspheres can act as microreactors to capture and degrade nanoplastics. Despite its lower enzymatic activity, the immobilized cutinase exhibits degradation performance comparable to that of its free enzyme counterpart. Moreover, it displays good operational stability over six degradation cycles. Therefore, our findings shed some light on achieving more efficient and cost-effective enzymatic nanoplastic remediation.
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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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
·
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
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DOI
PubMed
高影响因子
Catalysts
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2021
·Vol. 11
·期 10
·1158
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IF 4.146
Zhanyong Wang,Tingting Su,Jingjing Zhao
Genipin was used as a crosslinking agent to prepare magnetic genipin-crosslinked chitosan beads, which were then used as a carrier for immobilizing recombinant cutinase fromFusarium solani(FSC) to obtain immobilized FSC. The optimal temperature for the immobilized FSC was 55 °C, which was 5 °C higher than that of the free enzyme, whereas its optimal pH was increased from 8.0 to 9.0; this indicates that the immobilized FSC had improved pH and thermal stability. After repeated use for 10 cycles, the activity of the immobilized FSC remained at more than 50%; after being stored at 4 °C for 30 days, its activity was still approximately 88%. We also found that the Km of the immobilized FSC was higher than that of the free enzyme. These results indicate that the performance of FSC was improved after immobilization, which is an important basis for the subsequent application of FSC in industrial production.Keywords:immobilization;cutinase;chitosan;genipin
研究领域:
生物技术
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酶学
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材料科学
✓ 已验证
产品关联
DOI
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
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2021
·
IF 7.963
Qing-Song Huang,Zheng-Fei Yan,Xiao-Qian Chen,Yan-Yi Du,Juan Li,Zhan-Zhi Liu,Wei Xia,Sheng Chen,Jing Wu
Polyethylene terephthalate (PET) is a general plastic that produces a significant amount of waste due to its non-biodagradable properties. We obtained four bacteria ( Stenotrophomonas pavanii JWG-G1, Comamonas thiooxydans CG-1, Comamonas koreensis CG-2 and Fulvimonas soli GM-1) that utilize PET as a sole carbon source through a novel stepwise screening and verification strategy. PET films pretreated with S. pavanii JWG-G1 exhibited weight loss of 91.4% following subsequent degradation by Thermobifida fusca cutinase (TfC). S. pavanii JWG-G1 was able to colonize the PET surface and maintain high cell viability (over 50%) in biofilm, accelerating PET degradation. Compared with PET films with no pretreatment, pretreatment with S. pavanii JWG-G1 caused the PET surface to be significantly rougher with greater hydrophilicity (contact angle of 86.3 ± 2° vs. 96.6 ± 2°), providing better opportunities for TfC to contact and act on PET. Our study indicates that S. pavanii JWG-G1 could be used as a novel pretreatment for efficiently accelerating PET biodegradation by TfC.
研究领域:
环境科学
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微生物学
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Agriculture and Food Research
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2023
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IF 3.8
Meng Liu,Zihao Zong,Xiangjun Fang,Ruiling Liu,Honglei Mu,Hangjun Chen,Ben Niu,Haiyan Gao
An extracellular cutinase in Botrytis cinerea was isolated and its biochemical properties and effects on the penetration of pathogens were evaluated. This cutinase had a specific activity of 6.03 U/μg of protein after 22.33-fold purification, and yielded a recovery of 50.5%. The optimum pH and temperature for the enzymatic activity of cutinase were 9 and 45 °C, respectively. The cutinase was stable between 30 and 80 °C and pH 4.0–10.0. Fe 3+ and Cu 2+ strongly increased cutinase activity while Mn 2+ completely inhibited it. The enzyme could maintain its activity in the presence of methanol, ethanol and isopropanol, but was significantly inhibited in acetonitrile, n -hexane and acetone. Kinetic parameters K M , V max and k cat were 0.21 mM, 303 μmol•min −1 •mg −1 protein and 303.03 s −1 respectively. The purified cutinase could also destroy blueberry wax crystal and decrease the cuticle, indicating a positive role on the penetration of pathogens for blueberry fruits. This study provides useful information for fruit postharvest preservation.
研究领域:
植物病理学
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酶学
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食品科学
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产品关联
DOI
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2011
·Vol. 165
·期 2
·666-675
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IF 1.879
Chen Sheng,Liu Zhiguo,Chen Jian,Wu Jing
Escherichia coli is one of the most commonly used host strains for recombinant protein production. More and more research works on the production of recombinant protein indicate that extracellular production throughout a culture medium is more convenient and attractive compared to intracellular production. In present work, inducing temperature and isopropyl β- d -1-thiogalactopyranoside (IPTG) concentration were investigated to decrease the formation of inclusion body and increase the amount of soluble recombinant cutinase initially. Enzyme activity in the culture medium reached to 118.9 U/ml at 64 h of culture, and no inclusion body was detected in cytoplasm under the inducement condition of 0.2 mM IPTG and 30°C. In addition, it was found that a large amount of cutinase had been accumulated in periplasm since 16-h cultivation under the same inducement condition. Therefore, glycine and surfactant sodium taurodeoxycholate (TDOC) were further used to promote the leakage of recombinant cutinase from periplasm. Supplied with 100 mM glycine and 1 mM TDOC, the amount of cutinase in periplasm decreased remarkably, and the activity in the culture medium reached to 146.2 and 149.2 U/ml after 54 h of culturing, respectively.
研究领域:
生物工程
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分子生物学
✓ 已验证
DOI
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
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2022
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IF 8.025
Hang Yuan,Guanzhang Liu,Yaxin Chen,Zhiwei Yi,Wenhui Jin,Guangya Zhang
Enzymatic degradation of polyethylene terephthalate (PET) suffered from challenges such as complex and costly enzyme preparation , difficult access to PET substrates, poor reusability of free enzymes and sometimes MHET inhibitions. Herein, we propose an “all-in-one” strategy to address these issues with a well-designed elastin-like polypeptides (ELPs) tag. The preparation of the ELPs-tagged cutinase (ET-C) was efficient and easy to scale up by centrifugation , with an activity recovery of 57.55 % and a yield of 160 mg/L. Besides, the activity of the ET-C was 1.3 and 1.66-fold higher in degrading PET micro- and macro-plastics compared to wild-type cutinase. The self-immobilized cutinase (ET-C@SiO 2 ) obtained by the ELPs-mediated biosilicification exhibited high loading capacity, activity, and thermostability and maintained 77.65 % of the original activity after 10 reuses. Interestingly, the product of the ET-C was TPA, whereas the wild-type was TPA and MHET. This is a simple way to release the intermediates inhibition compared with the existing methods. Our results demonstrated the feasibility of the versatile ELPs tag, which will pave an alternative economic way for scalable PET biodegradation.
研究领域:
生物化学
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环境科学
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材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2019
·
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
BIORESOURCE TECHNOLOGY
·
2025
·
IF 9
Shiheng Chen,Lei Wang,Demin Kong,Zhisheng Wei,Xiaoqi Ling,Zhaohong Deng,Fengshan Zhang,Lingqia Su,Zhanzhi Liu,Jing Wu,Sheng Chen
Cutinase exhibits versatile biocatalytic potential in polymer degradation, textile processing, and industrial biocatalysis, where enhancing the thermal stability under extreme conditions is essential for practical applications. To enhance the thermal stability of Humicola insolens cutinase (HiC), a combination of strategies approach integrating error-prone PCR, computational design, and machine learning were implemented. Through systematic iterative recombination, an octuple mutant (M8) was developed with thermal tolerance and preserved catalytic activity. The engineered mutant demonstrated exceptional stability enhancements, exhibiting 8110- and 3982-fold increases in half-life at 65 °C and 70 °C, respectively, compared with wild-type HiC. Differential scanning calorimetry revealed a 13.0 °C elevation in melting temperature (T m ), while thermal inactivation analysis showed a 26.2 °C improvement in T (temperature causing 50 % activity loss in 60 min). Notably, M8 retained full activity at standard assay temperature and exhibited a broad pH profile despite incorporating eight stabilizing mutations. Molecular dynamics simulations and structural analyses revealed that the redistribution of surface electrostatic charges and a more compact overall structure were key factors in enhancing thermal stability. In summary, this study established a framework for rational thermostabilization of cutinases while providing molecular level insights into thermal adaptation mechanisms, with methodological implications for α/β-hydrolase family enzyme optimization. The engineered HiC mutant present significant potential for industrial processes requiring high-temperature operations.
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产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2024
·
IF 3.4
Yidi Liu,Zhanzhi Liu,Xuehong Guo,Ke Tong,Yueheng Niu,Zhiyu Shen,Hanzhi Weng,Fengshan Zhang,Jing Wu
The substantial accumulation of polyethylene terephthalate (PET) plastic waste in the environment has exacerbated the issue of plastic pollution. The biodegradation of PET plastics using biological enzymes has garnered considerable attention due to its efficiency and environmentally friendly nature. Nevertheless, the low binding affinity of PET plastics presents a significant limitation to the application of biocatalysts in their degradation. This study endeavors to engineer a fusion protein comprising the anchor peptide LCI, derived from Bacillus subtilis A014, and a thermally stabilized variant of Thermobifida fusca cutinase, D204C/E253C (Tfuc2), with the objective of augmenting its polyethylene terephthalate (PET) degradation efficacy. The findings demonstrate that LCI exhibits a high binding affinity for PET, and the hydrolytic efficiency of the LCI-containing fusion protein is enhanced by a factor of 1.8–34.5 compared to the free Tfuc2 enzyme. The enzymatic characteristics and molecular dynamics simulation outcomes indicate that the improved hydrolytic efficiency of PET may originate from the flexible oscillatory behavior of LCI, which exhibits a high binding affinity for PET. This study presents a novel methodology for the enzymatic degradation of PET plastic waste.
研究领域:
环境科学
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生物化学
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分子生物学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2024
·
IF 12.2
Zhitong Jiang,Xue Chen,Huizhen Xue,Zhoukun Li,Jinhui Lei,Muming Yu,Xin Yan,Hui Cao,Jie Zhou,Jiawei Liu,Mingna Zheng,Weiliang Dong,Yanwei Li,Zhongli Cui
Environmental pollution caused by plastic waste has become global problem that needs to be considered urgently. In the pursuit of a circular plastic economy, biodegradation provides an attractive strategy for managing plastic wastes, whereas effective plastic-degrading microbes and enzymes are required. In this study, we report that Blastobotrys sp. G-9 isolated from discarded plastic in landfills is capable of depolymerizing polyurethanes (PU) and poly (butylene adipate-co-terephthalate) (PBAT). Strain G-9 degrades up to 60% of PU foam after 21 days of incubation at 28 ℃ by breaking down carbonyl groups via secretory hydrolase as confirmed by structural characterization of plastics and degradation products identification. Within the supernatant of strain G-9, we identify a novel cutinase BaCut1, belonging to the esterase family, that can reproduce the same effect. BaCut1 demonstrates efficient degradation toward commercial polyester plastics PU foam (0.5 mg enzyme/25 mg plastic) and agricultural film PBAT (0.5 mg enzyme/10 mg plastic) with 50% and 18% weight loss at 37 ℃ for 48 h, respectively. BaCut1 hydrolyzes PU into adipic acid as a major end-product with 42.9% recovery via ester bond cleavage , and visible biodegradation is also identified from PBAT, which is a beneficial feature for future recycling economy. Molecular docking , along with products distribution, elucidates a special substrate-binding modes of BaCut1 with plastic substrate analogue . BaCut1-mediated polyester plastic degradation offers an alternative approach for managing PU plastic wastes through possible bio-recycling.
研究领域:
环境科学
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微生物学
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分子生物学
✓ 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）