来自Lycoris aurea的aldo-keto减少酶基因的功能性表征,这些基因参与了胺生物合成
Yiyu Xiang1, Xiaoran Jiang2, Jie Li2
1Nanjing University of Chinese Medicine, Nanjing 210023, China; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210014, China.
鉴定和描述了来自Lycoris aurea的两个aldo-keto减少酶 (AKR) 酶,LauAKR1和LauAKR2. 这些酶直接催化了加兰他胺生物合成中的关键降解步骤,这是一种有价值的植物代谢物.
科学领域:
- 植物生物化学 植物生物化学
- 酶学 是一种酶学.
- 代谢工程是代谢工程.
背景情况:
- 氨酸是一种植物性代谢物,具有显著的药理活性.
- 众所周知,阿尔多基因减少酶 (AKR) 基因家族参与植物二次代谢产物的生物合成.
- 了解负责胺生产的特定酶对于代谢工程和制药应用至关重要.
研究的目的:
- 识别和描述参与Lycoris物种兰胺生物合成的新型AKR酶.
- 阐明这些已识别的AKR同类的催化功能和局部化.
- 为特定AKR酶在减少narwedine到galantamine中的作用提供直接证据.
主要方法:
- 对AKR单基因的Lycoris转录组数据的选.
- 克隆和表征两种AKR同类物 (LauAKR1和LauAKR2) 来自金.
- 在各种压力条件下分析基因表达模式.
- 使用Nicotiana benthamiana中的短暂表达的亚细胞局部化研究.
- 在体外酶分析以确定基质特异性和辅因子依赖性.
主要成果:
- 确定了10个同源AKR单基因,并克隆和特征化了LauAKR1和LauAKR2.
- 这两种酶都属于AKR4B亚家族,并具有保存的活性部位残留物.
- AKR1和AKR2在金黄色菌组织中无处不在表达,在叶子,种子和球泡中含量更高.
- 基因表达是由NaCl诱导的,而不是甲基斯酸或曼尼托.
- 亚细胞定位证实LauAKR1和LauAKR2是细胞质的.
- 这两种酶都证明了脱甲基纳维丁和纳维丁的NADPH依赖性减少.
结论:
- AKR1和AKR2是催化剂的关键酶,在Lycoris物种中催化剂减少了narwedine到galantamine.
- 这些发现为AKR4B亚系成员在胺生物合成中的特定作用提供了直接证据.
- 描述的酶为代谢工程提供了潜在的目标,以增强胺的生产.
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