Journal Article

In vivo assembly in tobacco cells to elucidate and engineer the biosynthesis of 4-hydroxydihydrocinnamaldehyde from Gloriosa superba

Xiong, Zhiqiang, Jiang, Xuefei, Sun, Jingyi, Qiao, Fei, Ding, Zhuoying, Cong, Hanqing, Sun, Huapeng

2024

Overview

Key message This study described the biosynthesis of 4-hydroxydihydrocinnamaldehyde sharing with monolignol pathway and supplemented the biosynthesis of colchicine in G. superba , 4-hydroxydihydrocinnamaldehyde produced in tobacco BY2 cells provided an important stepstone. The precursor, 4-hydroxydihydrocinnamaldehyde (4-HDCA), participates in the biosynthesis of the carbon skeleton of colchicine, which is derived from L-phenylalanine. However, one hypothesis proposed that 4-HDCA is synthesized by sharing the early part of the monolignol pathway in G. superba . In this study, we validated this prediction and identified the enzymatic functions involved in this pathway. Gs DBR1 is a crucial enzyme to illustrate 4-HDCA diverging from monolignol pathway, we first confirmed its reductase activity on 4-coumaraldehyde, an important intermediate compound in monolignol biosynthesis. Then, the biochemical function of recombinant enzymes belonging to the other four families were verified to elucidate the entire process of 4-HDCA biosynthesis from L-phenylalanine . After reconstruction, the 4-HDCA was 78.4 ng/g with fresh weight (FW) of transgenic tobacco cells, and the yield increased to 168.22 ng/g·FW after improved treatment with methyl jasmonate (MeJA). The elucidation of 4-HDCA biosynthesis sharing the monolignol pathway supplemented the biosynthesis of colchicine in G. superba, and the production of 4-HDCA in tobacco cells provides an important step in the development of plant cell cultures as heterologous bio-factories for secondary metabolite production.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: Acetates - metabolism / Acetates - pharmacology / Aldehydes - metabolism / Biomedical and Life Sciences / Biosynthesis / Biotechnology / carbon / Cell Biology / Colchicine / Cyclopentanes - metabolism / Cyclopentanes - pharmacology / genetically modified organisms / Gloriosa superba / In vivo methods and tests / Industrial engineering / Life Sciences / Metabolites / Methyl jasmonate / Nicotiana - genetics / Nicotiana - metabolism / Original Article / oxidoreductases / Oxylipins - metabolism / Oxylipins - pharmacology / Phenylalanine / Phenylalanine - metabolism / Plant Biochemistry / Plant cells / Plant Proteins - genetics / Plant Proteins - metabolism / Plant Sciences / Plants, Genetically Modified / prediction / Reductases / secondary metabolites / Tobacco / Transgenic plants