The β-1,3-glucan synthase gene GFGLS2 plays major roles in mycelial growth and polysaccharide synthesis in Grifola frondosa

Jiang, Lihua; Li, Xiangfei; Zan, Xinyi; Fu, Xin; Cui, Fengjie; Zhu, Hong-An; Sun, Wenjing; Tao, Ting-Lei

doi:10.1007/s00253-021-11734-9

Cited by 15 publications

(20 citation statements)

References 42 publications

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“…The potential binding place for UDP-glucose in CMGLS is positioned in a pocket formed from α helixes α1, α2, and α3 and the beta sheet β1 (Figure A). The glucose and UDP moieties of the donor substrate bound the residues including His 871, Arg 876, Arg 1209, Tyr 1211, Ile 1212, Glu 1215, Glu 1220, Asp 1222, Gln 1229, Gln 1298, and Arg 1436 positioned in the predicted glucan synthase domain via hydrogen bonds, π-stacking, and salt bridges (Figure C), which provided a potential proof that the absence of QXXRW or R/KXGG motif in some β-1,3-glucan synthases still had the activity to catalyze the polymerization of the glucan chain. ,, …”

Section: Resultsmentioning

confidence: 93%

“…frondosa 9006-11 was wrongly annotated as a putative termination codon, which split the full sequence of β-1,3-glucan synthase genes into four sequences (GenBank: OBZ75851.1, OBZ75850.1). Accordingly, the β-1,3-glucan synthase genes GFGLS and GFGLS2 were corrected and reannotated with the full sequence of 5927 and 5944 bp (GenBank: MK80801.9 and MN477285.1). , …”

Section: Resultsmentioning

confidence: 99%

“…Similarly, no QXXRW or R/KXGG motif was found in the second glucan synthase (nominated as GFGLS2) of G. frondosa , cmFKS1 in C. militaris, and other FKS1p in A.…”

Section: Resultsmentioning

confidence: 99%

“…crassa having a molecular mass of >200 kDa with up to 16 transmembrane helices binding the substrate UDP-glucose Grifola frondosa contained at least two genes encoding β-1,3-glucan synthases GFGLSp and GFGLS2p with the predicted molecular masses of 203.66 and 195.2 kDa, respectively. , However, the functions and the catalytic characteristics/mechanisms of β-1,3-glucan synthases in fungi and mushrooms are still bottlenecked by their extremely complex structures with multi-transmembranes and large molecular masses. Saxena et al and Farkas et al pointed out that the catalysis of glucan synthases in yeasts and fungi usually depended on their interaction of the specific binding sites with the substrate UDP-glucose. , By searching for strictly conserved amino acid residues in the glycogen synthase of several species, the conserved sequence motif QXXRW or R/KXGG in glucan synthase of S.…”

Section: Introductionmentioning

confidence: 99%

“…16 In contrast, no QXXRW or R/KXGG motif was found in another G. frondosa glucan synthase (GFGLS2). 15 Cordyceps militaris is a highly valued edible/medicinal fungus and contains various polysaccharides with health benefits including antiproliferation of tumor cells, anti-inflammatory, and stimulation of immune system. 19 For instance, a linear β-1,3-glucan from fruiting bodies of C. militaris showed an antiinflammatory effect by the inhibition of IL-1β, TNF-α, and COX-2 expression and presented antinociceptive and antiinflammatory activities against formalin-induced nociception and LPS-induced peritonitis in mice.…”

Section: Introductionmentioning

confidence: 99%

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Functional Characterization and Structural Basis of the β-1,3-Glucan Synthase CMGLS from Mushroom Cordyceps militaris

Zan

Sun

et al. 2022

J. Agric. Food Chem.

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β-1,3-Glucan synthases play key roles in glucan synthesis, cell wall assembly, and growth of fungi. However, their multi-transmembrane domains (over 14 TMHs) and large molecular masses (over 100 kDa) significantly hamper understanding of their catalytic characteristics and mechanisms. In the present study, the 5841-bp gene CMGLS encoding the 221.7 kDa membranebound β-1,3-glucan synthase CMGLS in Cordyceps militaris was cloned, identified, and structurally analyzed. CMGLS was partially purified with a specific activity of 87.72 pmol/min/μg, a purification fold of 121, and a yield of 10.16% using a product-entrapment purification method. CMGLS showed a strict specificity to UDP-glucose with a K m value of 84.28 μM at pH 7.0 and synthesized β-1,3-glucan with a maximum degree of polymerization (DP) of 70. With the assistance of AlphaFold and molecular docking, the 3D structure of CMGLS and its binding features with substrate UDP-glucose were proposed for the first time to our knowledge. UDPglucose potentially bound to at least 11 residues via hydrogen bonds, π-stacking ,and salt bridges, and Arg 1436 was predicted as a key residue directly interacting with the moieties of glucose, phosphate, and the ribose ring on UDP-glucose. These findings would open an avenue to recognize and understand the glucan synthesis process and catalytic mechanism of β-1,3-glucan synthases in mushrooms.

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Section: Resultsmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

“…Similarly, no QXXRW or R/KXGG motif was found in the second glucan synthase (nominated as GFGLS2) of G. frondosa , cmFKS1 in C. militaris, and other FKS1p in A.…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functional Characterization and Structural Basis of the β-1,3-Glucan Synthase CMGLS from Mushroom Cordyceps militaris

Zan

Sun

et al. 2022

J. Agric. Food Chem.

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A newly discovered glycosyltransferase gene UGT88A1 affects growth and polysaccharide synthesis of Grifola frondosa

Li,

Wang,

Zhang

et al. 2024

Appl Microbiol Biotechnol

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Grifolafrodosa 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.

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Recent advances in the biosynthesis of fungal glucan structural diversity

Liu,

Hou,

Wang

et al. 2024

Carbohydrate Polymers

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The β-1,3-glucan synthase gene GFGLS2 plays major roles in mycelial growth and polysaccharide synthesis in Grifola frondosa

Cited by 15 publications

References 42 publications

Functional Characterization and Structural Basis of the β-1,3-Glucan Synthase CMGLS from Mushroom Cordyceps militaris

Functional Characterization and Structural Basis of the β-1,3-Glucan Synthase CMGLS from Mushroom Cordyceps militaris

A newly discovered glycosyltransferase gene UGT88A1 affects growth and polysaccharide synthesis of Grifola frondosa

Recent advances in the biosynthesis of fungal glucan structural diversity

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