食品科学 ›› 2024, Vol. 45 ›› Issue (17): 71-79.doi: 10.7506/spkx1002-6630-20231215-123

• 生物工程 • 上一篇    下一篇

亚油酸诱导植物乳杆菌p-8产生共轭亚油酸的蛋白组和类组蛋白乙酰化分析

李瑞华, 秦雅丽, 董其格其, 张和平, 赵国芬   

  1. (1.内蒙古农业大学生命科学学院,麦类种质创新利用自治区高等学校重点实验室,内蒙古 呼和浩特 010018;2.内蒙古农业大学食品科学与工程学院,内蒙古 呼和浩特 010018)
  • 出版日期:2024-09-15 发布日期:2024-09-09
  • 基金资助:
    内蒙古自然科学基金项目(2023MS03047;2023LHMS0); “饲用作物及有益微生物种质资源创新与分子育种”团队项目(TD202103); 内蒙古自治区研究生科研创新项目(S20231113Z)

Proteome and Histone-like Acetylation Analysis of Conjugated Linoleic Acid Production by Lactobacillus plantarum p-8 Induced by Linoleic Acid

LI Ruihua, QIN Yali, DONG Qigeqi, ZHANG Heping, ZHAO Guofen   

  1. (1. Key Lab of Germplasm Innovation and Utilization of Triticeae Crop at Universities of Inner Mongolia Autonomous Region, College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China;2. College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China)
  • Online:2024-09-15 Published:2024-09-09

摘要: 研究亚油酸(linoleic acid,LA)诱导下植物乳杆菌(Lactobacillus plantarum)p-8的蛋白组和类组蛋白乙酰化水平差异,探究共轭亚油酸(conjugated linoleic acid,CLA)产生的机制。结果表明,LA的最佳诱导质量浓度为1 mg/mL,在此质量浓度条件下,蛋白质组学分析发现,烯酰-酰基载体蛋白还原酶、酰基载体蛋白等脂肪酸合成的关键蛋白因子下调,脂肪酸合成下调,乙酰基转移酶水平上调。Western blot结合体外添加NaAc实验表明,乙酰辅酶A含量增加导致类组蛋白乙酰化水平提升。实时聚合酶链式反应检测显示,添加LA和NaAc使CLA相关酶、转录调控因子和sigma因子的mRNA表达升高,CLA含量也提高。相关性分析得出,乙酰辅酶A与LA水合酶转录水平和CLA含量之间呈显著正相关,揭示乙酰辅酶A和乙酰基转移酶含量上调导致类组蛋白乙酰化加强而上调CLA合成。结果为分子改良或科学调控乳酸菌以提高CLA转化率奠定了基础。

关键词: 亚油酸;共轭亚油酸;植物乳杆菌p-8;蛋白组;类组蛋白乙酰化

Abstract: In this study, changes in proteome and histone-like acetylation levels of Lactobacillus plantarum p-8 induced by linoleic acid (LA) were explored to investigate the mechanism of conjugated linoleic acid (CLA) production. The results showed that the optimal concentration of LA for inducing CLA production was 1 mg/mL. Proteomic analysis revealed that at this concentration, key protein factors for fatty acid synthesis such as enoyl acyl carrier protein (ACP) reductase and acyl carrier proteins were down-regulated, fatty acid synthesis was down-regulated, and acetyltransferase was up-regulated. Western blot showed that the increase in the content of acetyl-CoA upon NaAc addition resulted in elevated histone-like protein acetylation levels. The addition of LA and NaAc increased the mRNA expression of CLA-related enzymes, transcription factors (TF) and sigma factors, as determined by real-time polymerase chain reaction (real-time PCR), and increased the transformation rate of CLA. Correlation analysis yielded a significant positive correlation between the transcriptional levels of acetyl-CoA and linoleic acid hydratase (CLA-hy) and the transformation rate of CLA, revealing that the up-regulation of acetyl-CoA and acetyltransferase contents led to enhanced histone-like acetylation and consequent up-regulation of CLA synthesis. These results provide a basis for molecular modification and scientific regulation of lactic acid bacteria to improve CLA conversion.

Key words: linoleic acid; conjugated linoleic acid; Lactobacillus plantarum p-8; proteome; histone-like protein acetylation

中图分类号: