从Limosilactobacillus fermentum获得HcrF的生物化学表征,这是一个依赖NADH的2-还原酶,对氧酸有活性
1Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada, T6G 2P5.
Letters in applied microbiology
|November 9, 2024
概括
这项研究描述了HcrF,这是来自Limosilactobacillus fermentum的2-ene还原酶,对于在发酵食品中代谢化合物至关重要. HcrF有效地减少基胺酸,增加了已知参与植物化学转化的酶.
科学领域:
- 微生物学 微生物学
- 生物化学 生物化学
- 酶学 是一种酶学.
背景情况:
- 发酵植物食品中的化合物通过2 - 缩酶代谢.
- 乳酸细菌具有几种特征的2 - 烯还原酶,包括HcrB,Par1和一种daidzein还原酶.
- 了解这些酶是理解食品发酵中的植物化学转变的关键.
研究的目的:
- 从Limosilactobacillus fermentum中对HcrF,HcrB的同类进行表征.
- 为了确定HcrF的酶特性和基质特异性.
- 将HcrF置于其他2 - 烯还原酶的遗传学背景中.
主要方法:
- HcrF被克隆到大肠杆菌中,并使用亲和染色法进行净化.
- 在不同的温度和pH条件下评估了酶活性,并确定了辅助因子的要求.
- 通过测试各种酸的减少来确定基质特异性,并进行了植物遗传学分析.
主要成果:
- 在30°C-40°C和pH7.0之间,HcrF表现出最佳的活性,需要flavin mononucleotide和尼古丁胺胺氨基二核酸.
- 该酶有效地将ferulic,caffeic,p-coumaric和sinapic酸减少到它们的二衍生物,ferulic酸的Vmax最高.
- 遗传学分析显示HcrF与基胺酸减少酶HcrB密切结合,与Par1和黄/异黄减少酶不同.
结论:
- HcrF是一种来自Limosilactobacillus fermentum的新型酸缩酶,具有特定的基质偏好.
- 目前的蛋白质序列数据不足以预测二烯减少酶的基质特异性.
- 这项研究扩大了特征2-还原酶和它们在化合物代谢中的作用.
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