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新型 Zr-MOF/静电纺丝纳米纤维聚合物上的脂肪酶固定化:生化特性表征及高效生物柴油生产。

Lipase immobilization on a novel class of Zr-MOF/electrospun nanofibrous polymers: Biochemical characterization and efficient biodiesel production.

机构信息

Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.

Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.

出版信息

Int J Biol Macromol. 2021 Dec 1;192:1292-1303. doi: 10.1016/j.ijbiomac.2021.10.106. Epub 2021 Oct 20.

Abstract

In this study, due to the favorable properties of MOF compounds and fibrous materials, new nanostructures of Zr-MOF/PVP nanofibrous composites were synthesized by electrospinning procedure. The related features of these samples were characterized by relevant analyzes, including SEM, BET surface area analysis, XRD, and FTIR spectroscopy. The final product showed significant properties such as small particle size distribution, large surface area, and high crystallinity. This strategy for producing these nanostructures could lead to new compounds as novel alternative materials for biological applications. Lipase MG10 was successfully immobilized on the mentioned nanofibrous composites and biochemically characterized. The lipase activity of free and immobilized lipases was considered by measuring the absorbance of pNPP (500 μM in 40 mM Tris/HCl buffer, pH 7.8, and 0.01% Triton X100) at 37 °C for 30 min. Different concentrations of glutaraldehyde, different crosslinking times, different times of immobilization, different enzyme loading, and different pH values have been optimized. Results showed that the optimized immobilization condition was achieved in 2.5% glutaraldehyde, after 2 h of crosslinking time, after 6 h immobilization time, using 180 mg protein/g support at pH 9.0. The immobilized enzyme was also totally stable after 180 min incubation at 60 °C. The free enzyme showed the maximum activity at pH 9.0, but the optimal pH of the immobilized lipase was shifted about 1.5 pH units to the alkaline area. The immobilized lipase showed about 2.7 folds (78%) higher stability than the free enzyme at 50 °C. Some divalent metal ions, including Cu (22%), Co (37%), Mg (12%), Hg (11%), and Mn (17%) enhanced the enzyme activity of immobilized enzyme. The maximum biodiesel production (27%) from R. communis oil was obtained after 18 h of incubation by lipase MG10. The immobilized lipase displayed high potency in biodiesel production, about 83% after 12 h of incubation. These results indicated the high potency of Zr-MOF/PVP nanofibrous composites for efficient lipase immobilization.

摘要

在这项研究中,由于 MOF 化合物和纤维材料的优良特性,通过静电纺丝工艺合成了 Zr-MOF/PVP 纳米纤维复合材料的新型纳米结构。通过相关分析对这些样品的相关特征进行了表征,包括 SEM、BET 表面积分析、XRD 和 FTIR 光谱。最终产物表现出小颗粒尺寸分布、大表面积和高结晶度等显著特性。这种生产这些纳米结构的策略可能会导致新的化合物作为生物应用的新型替代材料。脂肪酶 MG10 成功地固定在上述纳米纤维复合材料上,并进行了生物化学表征。通过在 37°C 下测量 pNPP(40mM Tris/HCl 缓冲液,pH7.8,0.01%Triton X100 中的 500μM)的吸光度来考虑游离脂肪酶和固定化脂肪酶的酶活性持续 30 分钟。优化了不同浓度的戊二醛、不同的交联时间、不同的固定化时间、不同的酶载量和不同的 pH 值。结果表明,在 2.5%戊二醛、交联时间 2 小时、固定化时间 6 小时、pH9.0 时,固定化条件达到最佳。在 60°C 孵育 180 分钟后,固定化酶也完全稳定。游离酶在 pH9.0 时表现出最大活性,但固定化脂肪酶的最佳 pH 值向碱性区域移动约 1.5 pH 单位。固定化脂肪酶在 50°C 时的稳定性比游离酶高约 2.7 倍(78%)。一些二价金属离子,包括 Cu(22%)、Co(37%)、Mg(12%)、Hg(11%)和 Mn(17%),提高了固定化酶的酶活性。脂肪酶 MG10 作用 18 小时后,从 R.communis 油中获得了最大的生物柴油产量(27%)。固定化脂肪酶在生物柴油生产中表现出很高的效力,12 小时后约为 83%。这些结果表明 Zr-MOF/PVP 纳米纤维复合材料在有效固定脂肪酶方面具有很高的潜力。

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