Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i>
Bacterial infections pose a significant risk to human health. Magnolol, derived from <i>Magnolia officinalis</i>, exhibits potent antibacterial properties. Synthetic biology offers a promising approach to manufacture such natural compounds. However, the plant-based biosynthesis of magnol...
Main Authors: | , , , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2024-01-01
|
Series: | Molecules |
Subjects: | |
Online Access: | https://www.mdpi.com/1420-3049/29/3/587 |
_version_ | 1827354678168387584 |
---|---|
author | Yue Yang Zihe Li Hang Zong Shimeng Liu Qiuhui Du Hao Wu Zhenzhu Li Xiao Wang Lihui Huang Changlong Lai Meide Zhang Wen Wang Xianqing Chen |
author_facet | Yue Yang Zihe Li Hang Zong Shimeng Liu Qiuhui Du Hao Wu Zhenzhu Li Xiao Wang Lihui Huang Changlong Lai Meide Zhang Wen Wang Xianqing Chen |
author_sort | Yue Yang |
collection | DOAJ |
description | Bacterial infections pose a significant risk to human health. Magnolol, derived from <i>Magnolia officinalis</i>, exhibits potent antibacterial properties. Synthetic biology offers a promising approach to manufacture such natural compounds. However, the plant-based biosynthesis of magnolol remains obscure, and the lack of identification of critical genes hampers its synthetic production. In this study, we have proposed a one-step conversion of magnolol from chavicol using laccase. After leveraging 20 transcriptomes from diverse parts of <i>M. officinalis</i>, transcripts were assembled, enriching genome annotation. Upon integrating this dataset with current genomic information, we could identify 30 laccase enzymes. From two potential gene clusters associated with magnolol production, highly expressed genes were subjected to functional analysis. In vitro experiments confirmed MoLAC14 as a pivotal enzyme in magnolol synthesis. Improvements in the thermal stability of MoLAC14 were achieved through selective mutations, where E345P, G377P, H347F, E346C, and E346F notably enhanced stability. By conducting alanine scanning, the essential residues in MoLAC14 were identified, and the L532A mutation further boosted magnolol production to an unprecedented level of 148.83 mg/L. Our findings not only elucidated the key enzymes for chavicol to magnolol conversion, but also laid the groundwork for synthetic biology-driven magnolol production, thereby providing valuable insights into <i>M. officinalis</i> biology and comparative plant science. |
first_indexed | 2024-03-08T03:53:02Z |
format | Article |
id | doaj.art-c4a828603de14732ae546945aab9e401 |
institution | Directory Open Access Journal |
issn | 1420-3049 |
language | English |
last_indexed | 2024-03-08T03:53:02Z |
publishDate | 2024-01-01 |
publisher | MDPI AG |
record_format | Article |
series | Molecules |
spelling | doaj.art-c4a828603de14732ae546945aab9e4012024-02-09T15:18:43ZengMDPI AGMolecules1420-30492024-01-0129358710.3390/molecules29030587Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i>Yue Yang0Zihe Li1Hang Zong2Shimeng Liu3Qiuhui Du4Hao Wu5Zhenzhu Li6Xiao Wang7Lihui Huang8Changlong Lai9Meide Zhang10Wen Wang11Xianqing Chen12School of Ecology and Environment, Northwestern Polytechnical University, Xi’an 710072, ChinaSchool of Ecology and Environment, Northwestern Polytechnical University, Xi’an 710072, ChinaSchool of Ecology and Environment, Northwestern Polytechnical University, Xi’an 710072, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaSchool of Ecology and Environment, Northwestern Polytechnical University, Xi’an 710072, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaInstitute of Chinese Herbal Medicines, Hubei Academy of Agricultural Sciences, Enshi 445000, ChinaSchool of Ecology and Environment, Northwestern Polytechnical University, Xi’an 710072, ChinaJiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, ChinaBacterial infections pose a significant risk to human health. Magnolol, derived from <i>Magnolia officinalis</i>, exhibits potent antibacterial properties. Synthetic biology offers a promising approach to manufacture such natural compounds. However, the plant-based biosynthesis of magnolol remains obscure, and the lack of identification of critical genes hampers its synthetic production. In this study, we have proposed a one-step conversion of magnolol from chavicol using laccase. After leveraging 20 transcriptomes from diverse parts of <i>M. officinalis</i>, transcripts were assembled, enriching genome annotation. Upon integrating this dataset with current genomic information, we could identify 30 laccase enzymes. From two potential gene clusters associated with magnolol production, highly expressed genes were subjected to functional analysis. In vitro experiments confirmed MoLAC14 as a pivotal enzyme in magnolol synthesis. Improvements in the thermal stability of MoLAC14 were achieved through selective mutations, where E345P, G377P, H347F, E346C, and E346F notably enhanced stability. By conducting alanine scanning, the essential residues in MoLAC14 were identified, and the L532A mutation further boosted magnolol production to an unprecedented level of 148.83 mg/L. Our findings not only elucidated the key enzymes for chavicol to magnolol conversion, but also laid the groundwork for synthetic biology-driven magnolol production, thereby providing valuable insights into <i>M. officinalis</i> biology and comparative plant science.https://www.mdpi.com/1420-3049/29/3/587<i>Magnolia officinalis</i>transcriptsin vitromagnolol synthesisenzyme activity |
spellingShingle | Yue Yang Zihe Li Hang Zong Shimeng Liu Qiuhui Du Hao Wu Zhenzhu Li Xiao Wang Lihui Huang Changlong Lai Meide Zhang Wen Wang Xianqing Chen Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i> Molecules <i>Magnolia officinalis</i> transcripts in vitro magnolol synthesis enzyme activity |
title | Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i> |
title_full | Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i> |
title_fullStr | Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i> |
title_full_unstemmed | Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i> |
title_short | Identification and Validation of Magnolol Biosynthesis Genes in <i>Magnolia officinalis</i> |
title_sort | identification and validation of magnolol biosynthesis genes in i magnolia officinalis i |
topic | <i>Magnolia officinalis</i> transcripts in vitro magnolol synthesis enzyme activity |
url | https://www.mdpi.com/1420-3049/29/3/587 |
work_keys_str_mv | AT yueyang identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT ziheli identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT hangzong identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT shimengliu identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT qiuhuidu identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT haowu identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT zhenzhuli identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT xiaowang identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT lihuihuang identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT changlonglai identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT meidezhang identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT wenwang identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi AT xianqingchen identificationandvalidationofmagnololbiosynthesisgenesinimagnoliaofficinalisi |