Direct immobilization of Thermomyces lanuginosus lipase mediated by VKT-peptide for efficient biodiesel production from Jatropha curcas oil

doi:10.16085/j.issn.1000-6613.2023-0887

Abstract

Abstract:

Biodiesel is an environmentally friendly bio-liquid fuel. For the enzymatic production of biodiesel, there is a pressing need for cost-effective and efficient immobilized lipase. In this study, solid-bound peptide VKT-fused Thermomyces lanuginosus lipase (TLL) was directly immobilized on unmodified silica-containing materials to create a novel biocatalyst. Among the silica-based materials (ZSM-5, Na-Y, SAR-100, MCM-41 and SiO₂ powder) tested, TLL-VKT immobilized on ZSM-5 zeolite (TLL-VKT@ZSM-5) showed the best immobilization efficiency and maximum loading, as well as exceptional stability in terms of pH, temperature, storage and elution. When used as a biocatalyst at 40℃ for 48 hours, TLL-VKT@ZSM-5 (1%, w/w, oil) resulted in a biodiesel yield of 93.9% from Jatropha curcas oil. Furthermore, TLL-VKT@ZSM-5 exhibited high reusability, with a biodiesel yield of 71.9% after 7 repeated uses. The immobilization method in this study offers the advantages of simplicity, high efficiency, high stability, and reusability. In the context of enzyme immobilization used in the production of various chemicals, VKT peptides present significant potential.

Key words: Thermomyces lanuginosus lipase, immobilization, silica-containing materials, Jatropha curcas oil, biodiesel, SBPs

摘要：

生物柴油是一种环境友好的生物液体燃料，对于酶法生产生物柴油，迫切需要找到廉价和高效的固定化脂肪酶。本研究利用固体结合肽（SBPs）VKT，将其与疏棉状嗜热丝孢菌脂肪酶（Thermomyces lanuginosus lipase，TLL）融合构建融合脂肪酶，并将其固定化在硅基材料上，获得了一种新型的生物催化剂。在所测试的硅基材料（ZSM-5、Na-Y、SAR-100、MCM-41和SiO₂微粉）中，固定在ZSM-5沸石上的TLL-VKT（TLL-VKT@ZSM-5）表现出最佳的固定化效率和最大负载，且具有优异的pH、温度、储存和洗脱稳定性。以TLL-VKT@ZSM-5为生物催化剂，对麻疯树籽油进行转酯化反应，48h生物柴油得率即达到93.9%。此外，TLL-VKT@ZSM-5还表现出较高的重复使用性能，在7次重复使用后，生物柴油得率依然保持71.9%。本研究的酶固定化方法具有简单高效、稳定性高和重复使用性能高等优点。本研究显示VKT肽在酶蛋白固定化的应用方面具有较好前景。

关键词: 疏棉状嗜热丝孢菌脂肪酶, 固定化, 硅基材料, 麻疯树籽油, 生物柴油, 固体结合肽

CLC Number:

Q814.2

JI Xiaoyan, XU Rui, WANG Fei, LI Xun. Direct immobilization of Thermomyces lanuginosus lipase mediated by VKT-peptide for efficient biodiesel production from Jatropha curcas oil[J]. Chemical Industry and Engineering Progress, 2024, 43(6): 3285-3292.

季骁彦, 许蕊, 王飞, 李迅. VKT多肽介导的固定化疏棉状嗜热丝孢菌脂肪酶催化制备生物柴油[J]. 化工进展, 2024, 43(6): 3285-3292.

Figures/Tables 7

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洗脱剂	相对蛋白洗脱率/%
1mol/L NaCl	2.0±0.1
2mol/L NaCl	2.5±0.4
3mol/L NaCl	5.6±0.2
4mol/L NaCl	6.3±0.2
5mol/L NaCl	12.7±0.9
1% Tween-20	1.6±0.2
2% Tween-20	1.3±0.2
3% Tween-20	4.4±1.5
1% SDS	75.8±2.5
2% SDS	100.0±1.1
3% SDS	100.0±0.9

洗脱剂	相对蛋白洗脱率/%
1mol/L NaCl	2.0±0.1
2mol/L NaCl	2.5±0.4
3mol/L NaCl	5.6±0.2
4mol/L NaCl	6.3±0.2
5mol/L NaCl	12.7±0.9
1% Tween-20	1.6±0.2
2% Tween-20	1.3±0.2
3% Tween-20	4.4±1.5
1% SDS	75.8±2.5
2% SDS	100.0±1.1
3% SDS	100.0±0.9

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