|
|
|
| Metagenomic Mining of Functional Genes Related to Glycyrrhizin Synthesis from Endophytes of Licorice |
CHEN Ya-chao1,LI Nan-nan1,LIU Zi-di1,HU Bing1,**( ),LI Chun2 |
1 Institute of Biochemical Engineering, Department of Chemical Engineering, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, China
2 Key Lab for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China |
|
|
|
Abstract Objective:Licorice is widely used as a traditional Chinese medicine. Glycyrrhizin is the main active ingredient in licorice, a pentacyclic trterpenoid compound, has many pharmaceutical functions, such as anti-inflammatory, antiviral, and liver protection, and has been used in the clinical treatment of COVID-19. With the development of metabolic engineering and synthetic biology, the microbial synthesis of glycyrrhizin and its precursors has been gradually realized. However, the yield is low due to the incompatibility between plant genes and microbial chassis. In last decades, endophytes have been reported to harbor a wealth of functional traits, and could be directly or indirectly used for the production of plant-derived active compounds, indicating that investigation on endophytes has great academic potentials and application values in the field of metabolic engineering. Therefore, this study intends to conduct an in-depth study on the endophyte community of licorice and dig out the microbial-source functional genes that can be used for glycyrrhizin synthesis.Methods:In this study, three year old main root samples of Glycyrrhiza uralensis were collected from Emin County, Tacheng City, Xinjiang Uygur Autonomous Reyion. Metagenomic sequencing of endophytic communities was carried out, and the community structure and functional gene diversity of endophytic communities were analyzed. Through functional gene annotation and phylogenetic analysis, functional genes that may be involved in glycyrrhizin synthesis were excavated.Results:By analyzing the abundance of the community structure, it was found that the dominant endophytes in the licorice root were Steroidobacter denitrificans, Phenylobacterium zucineum, unclassified Phenylobacterium, and Phenylobacterium sp.. The metagenomic data were functionally annotated by COG database, KEGG database, and CAZy database, and the endophytic genes encoding enzymes involved in glycyrrhizin synthetic pathway, such as cytochrome P450 and UDP-glycosyltransferase, were found to be abundant in the endophytes of licorice.Conclusion:This study was the first to use metagenomic sequencing and analysis methods to understand the community structure and functional gene composition of endophytes from G. uralensis. It was proved that the endophytic community contained abundant cytochrome P450 and UGT coding genes, which laid a theoretical foundation for further comprehensive and in-depth study of the biological functions of endophytes from G. uralensis and how they transform into glycyrrhizin biosynthesis resources.
|
|
Received: 23 April 2021
Published: 30 September 2021
|
|
|
|
Corresponding Authors:
Bing HU
E-mail: binghu319@bit.edu.cn
|
|
|
| [1] |
Nomura Y, Seki H, Suzuki T, et al. Functional specialization of UDP -glycosyltransferase 73P12 in licorice to produce a sweet triterpenoid saponin, glycyrrhizin. The Plant Journal, 2019, 99(6):1127-1143.
doi: 10.1111/tpj.14409
pmid: 31095780
|
|
|
| [2] |
Zhao Y J, Lv B, Feng X D, et al. Perspective on biotransformation and de novo biosynthesis of licorice constituents. Journal of Agricultural and Food Chemistry, 2017, 65(51):11147-11156.
doi: 10.1021/acs.jafc.7b04470
|
|
|
| [3] |
Zhang L, Ren S C, Liu X F, et al. Mining of UDP-glucosyltrfansferases in licorice for controllable glycosylation of pentacyclic triterpenoids. Biotechnology and Bioengineering, 2020, 117(12):3651-3663.
doi: 10.1002/bit.v117.12
|
|
|
| [4] |
Matsui S, Matsumoto H, Sonoda Y, et al. Glycyrrhizin and related compounds down-regulate production of inflammatory chemokines IL-8 and eotaxin 1 in a human lung fibroblast cell line. International Immunopharmacology, 2004, 4(13):1633-1644.
doi: 10.1016/j.intimp.2004.07.023
|
|
|
| [5] |
Huang R Y, Chu Y L, Jiang Z B, et al. Glycyrrhizin suppresses lung adenocarcinoma cell growth through inhibition of thromboxane synthase. Cellular Physiology and Biochemistry, 2014, 33(2):375-388.
doi: 10.1159/000356677
|
|
|
| [6] |
Cinatl J, Morgenstern B, Bauer G, et al. Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus. The Lancet, 2003, 361(9374):2045-2046.
doi: 10.1016/S0140-6736(03)13615-X
|
|
|
| [7] |
Murck H. Symptomatic protective action of glycyrrhizin (licorice) in COVID-19 infection? Frontiers in Immunology, 2020, 11:1239. DOI: 10.3389/fimmu.2020.01239.
doi: 10.3389/fimmu.2020.01239
|
|
|
| [8] |
Bailly C, Vergoten G. Glycyrrhizin: an alternative drug for the treatment of COVID-19 infection and the associated respiratory syndrome? Pharmacology & Therapeutics, 2020, 214:107618.
|
|
|
| [9] |
Sun W T, Qin L, Xue H J, et al. Novel trends for producing plant triterpenoids in yeast. Critical Reviews in Biotechnology, 2019, 39(5):618-632.
doi: 10.1080/07388551.2019.1608503
|
|
|
| [10] |
Wang W, He P, Zhao D D, et al. Construction of Escherichia coli cell factories for crocin biosynthesis. Microbial Cell Factories, 2019, 18(1):1-11.
doi: 10.1186/s12934-018-1049-x
|
|
|
| [11] |
Gupta S, Chaturvedi P, Kulkarni M G, et al. A critical review on exploiting the pharmaceutical potential of plant endophytic fungi. Biotechnology Advances, 2020, 39:107462.
doi: 10.1016/j.biotechadv.2019.107462
|
|
|
| [12] |
Chen Y C, Hu B, Xing J M, et al. Endophytes: the novel sources for plant terpenoid biosynthesis. Applied Microbiology and Biotechnology, 2021, 105(11):4501-4513.
doi: 10.1007/s00253-021-11350-7
|
|
|
| [13] |
陈静, 许贞, 张雪, 等. 不同产地甘草内生真菌多样性及分离条件研究. 药学学报, 2019, 54(2):373-379.
|
|
|
| [13] |
Chen J, Xu Z, Zhang X, et al. Diversity and isolation parameters of endophytes from Glycyrrhiza uralensis of different habitats. Acta Pharmaceutica Sinica, 2019, 54(2):373-379.
|
|
|
| [14] |
Datta S, Rajnish K N, Samuel M S, et al. Metagenomic applications in microbial diversity, bioremediation, pollution monitoring, enzyme and drug discovery. A review. Environmental Chemistry Letters, 2020, 18(4):1229-1241.
doi: 10.1007/s10311-020-01010-z
|
|
|
| [15] |
Caparco A A, Pelletier E, Petit J L, et al. Metagenomic mining for amine dehydrogenase discovery. Advanced Synthesis & Catalysis, 2020, 362(12):2427-2436.
|
|
|
| [16] |
Carrión V J, Perez-Jaramillo J, Cordovez V, et al. Pathogen-induced activation of disease-suppressive functions in the endophytic root microbiome. Science, 2019, 366(6465):606-612.
doi: 10.1126/science.aaw9285
pmid: 31672892
|
|
|
| [17] |
Marshall J R, Yao P Y, Montgomery S L, et al. Screening and characterization of a diverse panel of metagenomic imine reductases for biocatalytic reductive amination. Nature Chemistry, 2021, 13(2):140-148.
doi: 10.1038/s41557-020-00606-w
pmid: 33380742
|
|
|
| [18] |
Mochida K, Sakurai T, Seki H, et al. Draft genome assembly and annotation of Glycyrrhiza uralensis, a medicinal legume. The Plant Journal, 2017, 89(2):181-194.
doi: 10.1111/tpj.13385
|
|
|
| [19] |
Kergourlay G, Taminiau B, Daube G, et al. Metagenomic insights into the dynamics of microbial communities in food. International Journal of Food Microbiology, 2015, 213:31-39.
doi: 10.1016/j.ijfoodmicro.2015.09.010
pmid: 26414193
|
|
|
| [20] |
宋素琴, 欧提库尔·玛合木提, 张志东, 等. 新疆胀果甘草内生菌的分离和鉴定. 微生物学通报, 2007, 34(5):867-870.
|
|
|
| [20] |
Song S Q, Otkur M, Zhang Z D, et al. Isolation and characterization of endophytic microorganisms in glaycyrrhiza inflat bat. from Xinjiang. Microbiology, 2007, 34(5):867-870.
|
|
|
| [21] |
李文军, 郑素慧, 毛培宏, 等. 新疆胀果甘草内生菌的分离及产甘草酸菌株的筛选. 生物技术, 2008, 18(5):28-31.
|
|
|
| [21] |
Li W J, Zheng S H, Mao P H, et al. Isolation of endophyte from Glycyrrhiza inflata batalin of Xinjiang uygur autonomous region and screening of glycyrrhizic acid-producing strain. Biotechnology, 2008, 18(5):28-31.
|
|
|
| [22] |
Tiwari P, Sangwan R S, Sangwan N S. Plant secondary metabolism linked glycosyltransferases: an update on expanding knowledge and Scopes. Biotechnology Advances, 2016, 34(5):714-739.
doi: 10.1016/j.biotechadv.2016.03.006
|
|
|
| [23] |
Girvan H M, Munro A W. Applications of microbial cytochrome P450 enzymes in biotechnology and synthetic biology. Current Opinion in Chemical Biology, 2016, 31:136-145.
doi: 10.1016/j.cbpa.2016.02.018
pmid: 27015292
|
|
|
| [24] |
Fiallos-Jurado J, Pollier J, Moses T, et al. Saponin determination, expression analysis and functional characterization of saponin biosynthetic genes in Chenopodium quinoa leaves. Plant Science, 2016, 250:188-197.
doi: S0168-9452(16)30094-2
pmid: 27457995
|
|
|
| [25] |
Morita M, Shibuya M, Kushiro T, et al. Molecular cloning and functional expression of triterpene synthases from pea (Pisum sativum). European Journal of Biochemistry, 2000, 267(12):3453-3460.
pmid: 10848960
|
|
|
| [26] |
Hayashi H, Huang P Y, Kirakosyan A, et al. Cloning and characterization of a cDNA encoding β-amyrin synthase involved in glycyrrhizin and soyasaponin biosyntheses in licorice. Biological and Pharmaceutical Bulletin, 2001, 24(8):912-916.
pmid: 11510484
|
|
|
| [27] |
Iturbe-Ormaetxe I, Haralampidis K, Papadopoulou K, et al. Molecular cloning and characterization of triterpene synthases from Medicago truncatula and Lotus japonicus. Plant Molecular Biology. 2003, 51(5):731-743.
pmid: 12683345
|
|
|
| [28] |
Zhu M, Wang C X, Sun W T, et al. Boosting 11-oxo-β-amyrin and glycyrrhetinic acid synthesis in Saccharomyces cerevisiae via pairing novel oxidation and reduction system from legume plants. Metabolic Engineering, 2018, 45:43-50.
doi: 10.1016/j.ymben.2017.11.009
|
|
|
| [29] |
Wilson A E, Tian L. Phylogenomic analysis of UDP-dependent glycosyltransferases provides insights into the evolutionary landscape of glycosylation in plant metabolism. The Plant Journal, 2019, 100(6):1273-1288.
doi: 10.1111/tpj.v100.6
|
|
|
| [30] |
Brazier-Hicks M, Gershater M, Dixon D, et al. Substrate specificity and safener inducibility of the plant UDP-glucose-dependent family 1 glycosyltransferase super-family. Plant Biotechnology Journal, 2018, 16(1):337-348.
doi: 10.1111/pbi.12775
pmid: 28640934
|
|
|
| [31] |
Chen K, Hu Z M, Song W, et al. Diversity of O-glycosyltransferases contributes to the biosynthesis of flavonoid and triterpenoid glycosides in Glycyrrhiza uralensis. ACS Synthetic Biology, 2019, 8(8):1858-1866.
doi: 10.1021/acssynbio.9b00171
|
|
|
| [32] |
Tiwari P, Bae H H. Horizontal gene transfer and endophytes: an implication for the acquisition of novel traits. Plants, 2020, 9(3):305.
doi: 10.3390/plants9030305
|
|
|
| [33] |
Tiwari R, Nain L, Labrou N E, et al. Bioprospecting of functional cellulases from metagenome for second generation biofuel production: a review. Critical Reviews in Microbiology, 2018, 44(2):244-257.
doi: 10.1080/1040841X.2017.1337713
|
|
|
| [34] |
Xin Y P, Guo T T, Zhang Y, et al. A new β-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity. Applied Microbiology and Biotechnology, 2019, 103(20):8439-8448.
doi: 10.1007/s00253-019-10092-x
|
|
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
