Yiting Zhu, Yinghui Feng, Jiawei Wang, Zhaoting Yuan, Yulu Miao, Tingwei Miao, Bei Gao* and Lujia Zhang*,
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引用次数: 0
摘要
通过脂肪酶催化的酯化作用合成长链 n-3 多不饱和脂肪酸(n-3 PUFA)三酰甘油(TAGs)对优化人体营养至关重要。然而,现有的脂肪酶对长链 n-3 PUFAs 缺乏选择性,阻碍了高度多不饱和 TAGs 的有效合成。通过二维核磁共振和量子化学计算,我们发现多不饱和 DHA 具有独特的弯曲构象。我们根据 DHA 的弯曲构象,利用虚拟饱和突变和结合自由能计算,合理设计了海洋链霉菌 W007 菌株的脂肪酶 MAS1。结果得到了氢键较多、与 DHA 的结合自由能(-28.3820 kcal/mol)较低的 D150V。D150V 对 DHA 的酯化选择性是野生型的 2.18 倍,而对山嵛酸的选择性则改善甚微。总之,我们的研究结果表明,脂肪酶的设计在选择性长链 n-3 PUFA 酯化方面取得了重大进展,从而为脂肪酶的改造和工业应用开辟了道路。
Selective Esterification Design of Lipases for TAG Synthesis Based on the Unique Structure of Curved DHA
The synthesis of long-chain n-3 polyunsaturated fatty acid (n-3 PUFA) triacylglycerols (TAGs) via lipase-catalyzed esterification is crucial for optimal human nutrition. However, existing lipases lack selectivity for long-chain n-3 PUFAs, hindering the efficient synthesis of highly polyunsaturated TAGs. By two-dimensional NMR and quantum chemical calculations, polyunsaturated DHA was found to have a unique curved conformation. We rationally designed the lipase MAS1 from marine Streptomyces sp. Strain W007 was based on the bent conformation of DHA, using virtual saturation mutagenesis and binding free energy calculations. Consequently, D150V with more hydrogen bonds and a lower binding free energy to DHA (−28.3820 kcal/mol) was obtained. The esterification selectivity of D150V to DHA was 2.18-fold that of the wild type, whereas its selectivity to behenic acid exhibited minimal improvement. Collectively, our findings present a significant advancement in lipase design for selective long-chain n-3 PUFA esterification, thereby opening avenues for lipase modification and industrial applications.