1. Effect of different immobilization conditions on the structure and activity of recombinant sn-1,3 lipase PEXANL1.
- Author
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Xie, Wei, Zhu, Ruonan, Long, Jia, Xing, Shuqi, Li, Cuiqin, Zeng, Xuefeng, and He, Laping
- Subjects
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IRON oxide nanoparticles , *IRON oxides , *IMMOBILIZED enzymes , *ASPERGILLUS niger , *ORGANIC solvents , *GLUTARALDEHYDE - Abstract
The study explored the effect of immobilizing PEXANL1, a recombinant Aspergillus niger sn -1,3 lipase, immobilization on chitosan-coated Fe 3 O 4 magnetic nanoparticles (Fe 3 O 4 @CS MNPs) on its activity and secondary structure. Initially, Fe 3 O 4 @CS MNPs were synthesized and employed to facilitate the covalent immobilization of PEXANL1, confirmed through SEM, XRD, FTIR, VSM, XRD, and TGA analyses. The PEXANL1-Fe 3 O 4 @CS MNPs demonstrated significantly enhanced performance, achieving a maximum activity recovery of 154 % ± 2 % and an immobilization efficiency of 72 % ± 0.8 % under optimum conditions: acetonitrile + acetone as precipitant, support mass of 125 mg, a precipitant to enzyme solution ratio 4:1, glutaraldehyde concentration of 15 mM, crosslinking time of 1 h. Relative to PEXANL1, the immobilized enzyme showed a 17.2 % reduction in β-fold content. In contrast, α-helix, β-turn, and irregular curl contents increased by 9.3 %, 0.75 %, and 6.74 %, respectively, indicating a more ordered structural composition and enhanced stability. Repeated use of PEXANL1-Fe 3 O 4 @CS MNPs maintained about 52 % of its initial activity after 8 cycles. Glycerolysis tests showed that the sn -1,3 specificity of the immobilized PEXANL1 was 71.15 %. Compared to PEXANL1, immobilized PEXANL1 exhibited more excellent to variations in temperature, pH, metal ions, and organic solvents. [Display omitted] • The activity recovery of immobilized PEXANL1 under optimal conditions was 154 %. • Immobilization improves the ordered structure in the secondary structure of PEXANL1. • Immobilized PEXANL1 remains the sn -1,3 selectivity to triolein. • Immobilization improves the enzymatic properties and operational stability of PEXANL1. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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