功能性特征和催化活性改善的玻利乙烯转移酶从长 (Wurfbainia longiligularis) 的
Yuanxia Chen1, Tiantian Wang1, Huilin Liang1
1Key Laboratory of Chinese Medicinal Resource from Lingnan (Ministry of Education), School of Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Journal of agricultural and food chemistry
|May 30, 2024
概括
研究人员在合成酸 (BA) 的植物中确定了三种酸转移酶 (BAT). WlBAT1显示出高效率,并且特定的突变增强了其用于代谢工程和合成生物学应用的活性.
科学领域:
- 植物生物化学和分子生物学
- 二次代谢物生物合成二次代谢物生物合成
- 酶催化和工程 酶催化和工程
背景情况:
- 在植物的二次代谢物生产中,乙基化非常重要,BAHD乙基转移酶是关键参与者.
- 玻利乙烯转移酶 (BAT) 从玻利和乙-CoA中合成玻利乙酸盐 (BA).
- 之前对*Wurfbainia villosa*中BA生物合成的研究仅限于*体外*工作,具有较低的催化活性.
研究的目的:
- 为了识别和描述编码BAT的基因,参与酸生物合成.
- 调查已识别的BAT的基质特异性和催化效率.
- 为潜在应用阐明影响 BAT 活动的关键残留物和突变.
主要方法:
- 基因鉴定和克隆三个假定BAT基因 (WlBAT1,WlBAT2,WlBAT3).
- *体外*酶测试以确定催化活性和基质特异性.
- 分子模拟和局部定向突变发生,以确定关键残留物和工程酶变体.
主要成果:
- 确定了WlBAT1,WlBAT2和WlBAT3三种基因,并表明它们编码了功能性BAT *in vitro*.
- 对于玻利类基板来说,WlBAT1的催化效率最高.
- 虽然BAT显示了广泛的基质耐受性 *in vitro*,但它们是针对波醇类基质的特定 *in vivo*. 确定了关键的残留物 (D32,N36,H168,N297,N355,H384),一个双突变物 (R382I-D385R) 提高了化效率.
结论:
- WlBAT1是一种高效的酶,用于乙酸的合成.
- 特定的氨基酸残留对WlBAT1的催化功能至关重要.
- 工程化BAT变种为代谢工程和合成生物学提供了生产酸的潜力.
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