A Newly Discovered Glycosyltransferase Gene UGT88A1 Affects Growth and Polysaccharide Synthesis of Grifola Frondosa

Overview

Journal Appl Microbiol Biotechnol

Specialties Biomedical Engineering
Biotechnology
Microbiology

Date 2024 Feb 29

PMID 38421403

Authors

Jian Li

Bao-Xin Wang

Jie Zhang

Na Han

Shu-Ting Liu

Wen-Ji Geng

Shi-Ru Jia

Yan-Ru Li

Quan Gan

Pei-Pei Han

Affiliations

Soon will be listed here.

Abstract

Grifola frodosa polysaccharides, especially β-D-glucans, possess significant anti-tumor, antioxidant and immunostimulatory activities. However, the synthesis mechanism remains to be elucidated. A newly discovered glycosyltransferase UGT88A1 was found to extend glucan chains in vitro. However, the role of UGT88A1 in the growth and polysaccharide synthesis of G. frondosa in vivo remains unclear. In this study, the overexpression of UGT88A1 improved mycelial growth, increased polysaccharide production, and decreased cell wall pressure sensitivity. Biomass and polysaccharide production decreased in the silenced strain, and the pressure sensitivity of the cell wall increased. Overexpression and silencing of UGT88A1 both affected the monosaccharide composition and surface morphology of G. frondosa polysaccharides and influenced the antioxidant activity of polysaccharides from different strains. The messenger RNA expression of glucan synthase (GLS), UTP-glucose-1-phosphate uridylyltransferase (UGP), and UDP-xylose-4-epimerase (UXE) related to polysaccharide synthesis, and genes related to cell wall integrity increased in the overexpression strain. Overall, our study indicates that UGT88A1 plays an important role in the growth, stress, and polysaccharide synthesis of G. frondosa, providing a reference for exploring the pathway of polysaccharide synthesis and metabolic regulation. KEY POINTS: •UGT88A1 plays an important role in the growth, stress response, and polysaccharide synthesis in G. frondosa. •UGT88A1 affected the monosaccharide composition, surface morphology and antioxidant activity of G. frondosa polysaccharides. •UGT88A1 regulated the mRNA expression of genes related to polysaccharide synthesis and cell wall integrity.

References

Li M, Chen T, Gao T, Miao Z, Jiang A, Shi L . UDP-glucose pyrophosphorylase influences polysaccharide synthesis, cell wall components, and hyphal branching in Ganoderma lucidum via regulation of the balance between glucose-1-phosphate and UDP-glucose. Fungal Genet Biol. 2015; 82:251-63. DOI: 10.1016/j.fgb.2015.07.012. View

Lo T, Jiang Y, Chao A, Chang C . Use of statistical methods to find the polysaccharide structural characteristics and the relationships between monosaccharide composition ratio and macrophage stimulatory activity of regionally different strains of Lentinula edodes. Anal Chim Acta. 2007; 584(1):50-6. DOI: 10.1016/j.aca.2006.10.051. View

Cui F, Wu X, Tao T, Zan X, Sun W, Mu D . Functions of a Glucan Synthase Gene in Mycelial Growth and Polysaccharide Production of . J Agric Food Chem. 2019; 67(32):8875-8883. DOI: 10.1021/acs.jafc.9b03569. View

Romano N, Macino G . Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences. Mol Microbiol. 1992; 6(22):3343-53. DOI: 10.1111/j.1365-2958.1992.tb02202.x. View

Su Y, Li L . Structural characterization and antioxidant activity of polysaccharide from four auriculariales. Carbohydr Polym. 2019; 229:115407. DOI: 10.1016/j.carbpol.2019.115407. View

Jiang L, Li X, Zan X, Fu X, Cui F, Zhu H . The β-1,3-glucan synthase gene GFGLS2 plays major roles in mycelial growth and polysaccharide synthesis in Grifola frondosa. Appl Microbiol Biotechnol. 2021; 106(2):563-578. DOI: 10.1007/s00253-021-11734-9. View

Krylov V, Nifantiev N . Synthetic Oligosaccharides Mimicking Fungal Cell Wall Polysaccharides. Curr Top Microbiol Immunol. 2019; 425:1-16. DOI: 10.1007/82_2019_187. View

Han P, Sun Y, Jia S, Zhong C, Tan Z . Effects of light wavelengths on extracellular and capsular polysaccharide production by Nostoc flagelliforme. Carbohydr Polym. 2014; 105:145-51. DOI: 10.1016/j.carbpol.2014.01.061. View

Wang X, Zhang L, Chen N, Li J, Han C, Wang S . The effects of quorum sensing molecule farnesol on the yield and activity of extracellular polysaccharide from Grifola frondosa in liquid fermentation. Int J Biol Macromol. 2021; 191:377-384. DOI: 10.1016/j.ijbiomac.2021.09.088. View

10.

Rautengarten C, Birdseye D, Pattathil S, McFarlane H, Saez-Aguayo S, Orellana A . The elaborate route for UDP-arabinose delivery into the Golgi of plants. Proc Natl Acad Sci U S A. 2017; 114(16):4261-4266. PMC: 5402404. DOI: 10.1073/pnas.1701894114. View

11.

Punt P, Oliver R, Dingemanse M, Pouwels P, van den Hondel C . Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli. Gene. 1987; 56(1):117-24. DOI: 10.1016/0378-1119(87)90164-8. View

12.

Cescutti P, Kallioinen A, Impallomeni G, Toffanin R, Pollesello P, Leisola M . Structure of the exopolysaccharide produced by Enterobacter amnigenus. Carbohydr Res. 2005; 340(3):439-47. DOI: 10.1016/j.carres.2004.12.008. View

13.

Dichtl K, Helmschrott C, Dirr F, Wagener J . Deciphering cell wall integrity signalling in Aspergillus fumigatus: identification and functional characterization of cell wall stress sensors and relevant Rho GTPases. Mol Microbiol. 2012; 83(3):506-19. DOI: 10.1111/j.1365-2958.2011.07946.x. View

14.

Zan X, Wu X, Cui F, Zhu H, Sun W, Jiang L . UDP-glucose pyrophosphorylase gene affects mycelia growth and polysaccharide synthesis of Grifola frondosa. Int J Biol Macromol. 2020; 161:1161-1170. DOI: 10.1016/j.ijbiomac.2020.06.139. View

15.

Shen S, Jia S, Wu Y, Yan R, Lin Y, Zhao D . Effect of culture conditions on the physicochemical properties and antioxidant activities of polysaccharides from Nostoc flagelliforme. Carbohydr Polym. 2018; 198:426-433. DOI: 10.1016/j.carbpol.2018.06.111. View

16.

Zhang X, Jia X, Tian S, Zhang C, Lu Z, Chen Y . Role of the small GTPase Rho1 in cell wall integrity, stress response, and pathogenesis of Aspergillus fumigatus. Fungal Genet Biol. 2018; 120:30-41. DOI: 10.1016/j.fgb.2018.09.003. View

17.

Zan X, Zhu H, Jiang L, Liang Y, Sun W, Tao T . The role of Rho1 gene in the cell wall integrity and polysaccharides biosynthesis of the edible mushroom Grifola frondosa. Int J Biol Macromol. 2020; 165(Pt A):1593-1603. DOI: 10.1016/j.ijbiomac.2020.09.239. View

18.

Chen X, Wang Q, Liu N, Liu G, Chi Z, Chi Z . A glycosyltransferase gene responsible for pullulan biosynthesis in Aureobasidium melanogenum P16. Int J Biol Macromol. 2016; 95:539-549. DOI: 10.1016/j.ijbiomac.2016.11.081. View

19.

Ji S, Liu R, Ren M, Li H, Xu J . Enhanced Production of Polysaccharide Through the Overexpression of Homologous Uridine Diphosphate Glucose Pyrophosphorylase Gene in a Submerged Culture of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Higher Basidiomycetes). Int J Med Mushrooms. 2015; 17(5):435-42. DOI: 10.1615/intjmedmushrooms.v17.i5.30. View

20.

Mao G, Zou Y, Feng W, Wang W, Zhao T, Ye C . Extraction, preliminary characterization and antioxidant activity of Se-enriched Maitake polysaccharide. Carbohydr Polym. 2013; 101:213-9. DOI: 10.1016/j.carbpol.2013.09.034. View