Frontiers in Bioengineering and Biotechnology
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2022
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IF 6.064
Liu Yu,Liu Chen,Liu Huan,Zeng Qi,Tian Xinpeng,Long Lijuan,Yang Jian
Poly (ethylene terephthalate) (PET) plastic is chemically inert and persistent. Massive quantities of PET waste end up in landfill sites and oceans, posing major global pollution concerns. PET degrading enzymes with high efficiency provide plastic recycling and bioremediation possibilities. Here, we report a novel cutinase, MtCut with distinct catalytic behaviors, derived from the deep sea Nocardiopsaceae family strain. Biochemical analyses showed MtCut efficiently hydrolyzed PET at ambient temperatures and in an exo-type manner. The activity and stability of MtCut were enhanced by the addition of calcium ions. Notably, no hydrolysis products inhibition was observed during PET depolymerization, suggesting MtCut is a better biocatalyst when compared to other PET hydrolases. In addition, structural components associated with thermal adaptation were investigated using molecular dynamic (MD) simulations, and key regions regulating MtCut thermostability were identified. Our biochemical and structural analyses of MtCut deepen the understanding of PET hydrolysis by cutinases, and provide invaluable insights on improvement and performance engineering strategies for PET-degrading biocatalysts.
研究领域:
生物化学
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环境科学
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微生物学
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酶学
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生物技术
✓ 已验证
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2022
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IF 4.446
Lankai Su,Kun Chen,Shu Bai,Linling Yu,Yan Sun
Environmental-friendly treatment of poly (ethylene terephthalate) (PET) is important for the circular plastic economy, and enzymatic degradation provided a promising approach. However, enzyme binding toward the PET surface is a rate-limiting factor in PET hydrolysis . Thus, a 16-residue peptide (αSP) from the C-terminal sequence of α-synuclein was used to fuse to the C-terminal of a variant (ICCG) of the leaf-branch compost cutinase , and supposed to serve as an anchor peptide for PET degradation. The fusion enzyme (ICCG-αSP) was characterized to keep the same secondary and tertiary structures as ICCG, but enhanced enzyme binding onto bis(2-hydroxyethyl) terephthalate and PET, respectively. Moreover, αSP-fusion enhanced enzyme expression by over 50%, and increased the catalytic efficiency of p -nitrophenyl butyrate by 20% without compromising the enzyme stability . For PET degradation, ICCG-αSP was 3.89 times more active than ICCG by 3-h reaction at 50 °C, and exhibited 1.05–1.45 times higher degradation activity than ICCG in 4-d reaction at 50–70 °C, and pH 7.0 to pH 9.0, respectively. Furthermore, αSP-fusion lowered the optimal long-term degradation temperature from 65 °C to 50 °C at pH 8.0. Therefore, αSP-fusion was effective to increase the hydrolytic efficiency of PET hydrolases by increasing the degradation efficiency and decreasing the degradation temperature.
研究领域:
环境科学
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生物技术
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酶学
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高分子科学
✓ 已验证
DOI
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
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Jie Wang,Jin Xu,Jiugang Yuan,Xuerong Fan
Water-dispersed polyester (WPET) has irreplaceable and wide application prospects in the textile field, but it is easy to form printing and dyeing wastewater during traditional fabric treatment. In this paper, cutinase-catalyzed green hydrolysis of WPET was achieved. The main purpose of this paper is to study the difference between two PEG-free WPETs with different -SO 3 − content in the cutinase-catalyzed hydrolysis process. Differences in solution properties, enzyme activity, SSIPA, and TPA release as well as functional groups and thermal properties of the hydrolysis products of the systems during cutinase hydrolysis were analyzed in detail for 14.1 % S-0 % PEG and 9.5 % S-0 % PEG. Results show that the structure of WPET has an influential effect on its hydrolysis rate in cutinase-catalyzed systems, which mainly originates from the -SO 3 − content. SSIPA released by WPET during hydrolysis has an inhibitory effect on cutinase enzyme activity. The hydrolysis process of WPET with higher -SO 3 − content releases more SSIPA, which in turn inhibits the enzyme activity of cutinase in the reaction system, ultimately leading to a slower hydrolysis rate of WPET. These findings provide new ideas for cutinase to target WPETs with different structures and are important for the green development of the textile industry.
研究领域:
生物化学
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纺织科学
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环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION
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2023
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IF 1.9
Ting Hu,Yujun Wang,Jingwei Lin,Zhanyong Wang
Poly(butylene succinate) (PBS), poly(hexamethylene succinate) (PHS) and three kinds of poly(butylene succinate-co-hexamethylene succinate)s (P(BS-co-HS)s) were enzymatically hydrolyzed by Fusarium solani cutinase. The results showed that the characteristics of the enzymatic hydrolysis of these polyesters were mainly affected by crystallinity, thermal properties and the BS/HS ratio. The enzymatic hydrolysis rates of the polyesters are as follows: P(BS-co-HS52) ≈ P(BS-co-HS71) > PHS > P(BS-co-HS32) > PBS. Furthermore, with increasing HS content, both the degree of crystallinity (Xc) and melting temperature (Tm) of the polyesters first decreased and then increased. P(BS-co-HS52) and P(BS-co-HS71) had the lowest Xc and the lowest Tm, thus had the highest hydrolysis rate; this shows that the hydrolysis rate is affected by Xc and Tm. The results also showed that BS/HS ratio could affect the physical properties and degradability of polyesters. Thus, it is possible to prepare polyesters with various physical properties and degradability for different applications by adjusting BS/HS ratio. The crystalline and amorphous regions of the polyesters were both hydrolyzed, during which parts of the crystalline regions were converted into the amorphous regions. Finally, we found that the crystal structure and thermal stability of the polyesters were not affected by the enzymatic hydrolysis.
研究领域:
生物化学
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材料科学
✓ 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）