Wurfbainia longiligularis Borneol Acetyltransferase 的功能表征和催化活性改进。

IF 6.2 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Yuanxia Chen, Tiantian Wang, Huilin Liang, Dongming Ma, Ruoting Zhan*, Jinfen Yang* and Peng Yang*, 
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引用次数: 0

摘要

在植物次生代谢物的生物合成过程中,酰化是一个多样化的生理过程,其中 BAHD 乙酰转移酶发挥着重要作用。龙脑醇乙酰转移酶(BAT)是一种醇乙酰转移酶,可催化龙脑醇和乙酰-CoA 合成乙酸龙脑酯(BA)。然而,迄今为止,参与 BA 生物合成的酶只在 Wurfbainia villosa 中得到表征,对 WvBAT 的研究也只在体外进行,而且催化活性相对较低。本研究确定了三个基因(WlBAT1、WlBAT2 和 WlBAT3),它们编码的 BATs 能够在体外乙酰化龙脑醇以合成 BA。我们还确定,WlBAT1 对冰片醇类底物(包括 (+)- 冰片醇、(-)-冰片醇和异冰片醇)的催化效率最高。此外,我们还发现 BATs 在体外可以催化多种底物类型,但在体内只能催化龙脑醇类底物。通过分子模拟和定点突变,我们发现残基 D32、N36、H168、N297、N355 和 H384 对 WlBAT1 的催化活性至关重要,而 WlBAT1 的 R382I-D385R 双突变体在体外和体内对冰片醇类底物的酰化效率都在增加。这些发现为植物代谢工程和合成生物学生产 BA 提供了关键的遗传要素。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Functional Characterization and Catalytic Activity Improvement of Borneol Acetyltransferase from Wurfbainia longiligularis

Functional Characterization and Catalytic Activity Improvement of Borneol Acetyltransferase from Wurfbainia longiligularis

Functional Characterization and Catalytic Activity Improvement of Borneol Acetyltransferase from Wurfbainia longiligularis

In plant secondary metabolite biosynthesis, acylation is a diverse physiological process, with BAHD acyltransferases playing an essential role. Borneol acetyltransferase (BAT) is an alcohol acetyltransferase, which catalyzes borneol and acetyl-CoA to synthesize bornyl acetate (BA). However, the enzymes involved in the biosynthesis of BA have so far only been characterized in Wurfbainia villosa, the studies on the WvBATs have only been conducted in vitro, and the catalytic activity was relatively low. In this research, three genes (WlBAT1, WlBAT2, and WlBAT3) have been identified to encode BATs that are capable of acetylating borneol to synthesize BA in vitro. We also determined that WlBAT1 has the highest catalytic efficiency for borneol-type substrates, including (+)-borneol, (−)-borneol, and isoborneol. Furthermore, we found that BATs could catalyze a wide range of substrate types in vitro, but in vivo, they exclusively catalyzed borneol-type substrates. Through molecular simulations and site-directed mutagenesis, it was revealed that residues D32, N36, H168, N297, N355, and H384 are crucial for the catalytic activity of WlBAT1, while the R382I-D385R double mutant of WlBAT1 exhibited an increasing acylation efficiency for borneol-type substrates in vitro and in vivo. These findings offer key genetic elements for the metabolic engineering of plants and synthetic biology to produce BA.

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来源期刊
Journal of Agricultural and Food Chemistry
Journal of Agricultural and Food Chemistry 农林科学-农业综合
CiteScore
9.90
自引率
8.20%
发文量
1375
审稿时长
2.3 months
期刊介绍: The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.
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