Combined fermentation and ARTP mutagenesis to enhance lipase activity of Penicillium camembertii and its application for high-purity 1,3-diacylglycerol preparation

IF 8.5 1区农林科学 Q1 CHEMISTRY, APPLIED

Food Chemistry Pub Date : 2024-11-16 DOI:10.1016/j.foodchem.2024.142072

Shulan Ji, Hong Yang, Xinyu Xie, Yu Zhang, Xun Li, Fei Wang

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引用次数: 0

Abstract

Penicillium camembertii lipase (PCL) is a glycerol-biased enzyme isolated from.P. camembertii, exhibiting high esterification activity. The PCL activity was enhanced by combining fermentation optimization with atmospheric and room temperature plasma (ARTP) mutagenesis. Following multiple rounds of high-throughput screening and serial passages, a genetically stable mutant, P12, was obtained. In shake flask culture, P12 exhibited a lipase activity of 1600 U/g, representing an 800 % increase compared to the wild type (WT), with improved thermostability and methanol stability. Subsequently, P12 was used as a whole-cell biocatalyst to catalyze the esterification of oleic acid with glycerol in a solvent-free system to prepare 1,3-diacylglycerol (1,3-DAG). With an oleic acid /glycerol molar ratio of 4:1, 8 % biocatalyst, and a reaction temperature of 40 °C, the content of 1,3-DAG was 74.7 % after 24 h. These results suggest that the mutant P12 demonstrates considerable potential as a whole-cell biocatalyst for synthesizing high-purity 1,3-DAG.

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联合发酵和 ARTP 诱变提高卡门贝氏青霉脂肪酶活性及其在制备高纯度 1,3-二酰甘油中的应用

卡门培尔青霉脂肪酶（PCL）是从卡门培尔青霉中分离出来的一种甘油基酶，具有很高的酯化活性。通过将发酵优化与大气和室温等离子体（ARTP）诱变相结合，提高了 PCL 的活性。经过多轮高通量筛选和连续传代，获得了基因稳定的突变体 P12。在摇瓶培养中，P12 表现出 1600 U/g 的脂肪酶活性，与野生型（WT）相比提高了 800%，其热稳定性和甲醇稳定性也得到了改善。随后，P12 被用作全细胞生物催化剂，在无溶剂体系中催化油酸与甘油的酯化反应，制备出 1,3-二酰甘油（1,3-DAG）。在油酸/甘油摩尔比为 4:1、生物催化剂含量为 8%、反应温度为 40 ℃的条件下，24 小时后 1,3-DAG 的含量为 74.7%。这些结果表明，突变体 P12 作为合成高纯度 1,3-DAG 的全细胞生物催化剂具有相当大的潜力。

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来源期刊

Food Chemistry 工程技术-食品科技

CiteScore

16.30

自引率

10.20%

发文量

3130

审稿时长

122 days

期刊介绍： Food Chemistry publishes original research papers dealing with the advancement of the chemistry and biochemistry of foods or the analytical methods/ approach used. All papers should focus on the novelty of the research carried out.