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...

Full description

Bibliographic Details
Main Authors: 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
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