4.7 Article

Increased production and anti-senescence activity of exopolysaccharides in Ganoderma lingzhi by co-overexpression of β-1,3-glucan synthase and UDP-glucose pyrophosphorylase

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DOI: 10.1016/j.ijbiomac.2023.126778

Keywords

Polysaccharide; Fermentation; Genetic engineering; Glucan synthase; Biosynthesis; Anti-senescence

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A β-1,3-glucan synthase gene was cloned and overexpressed in Ganoderma lingzhi, leading to an increase in intra-cellular polysaccharides content. Simultaneous overexpression of this gene and UDP-glucose pyrophosphorylase gene further enhanced the production of polysaccharides, with higher glucose content and improved antioxidative and anti-senescence activities.
A beta-1,3-glucan synthase gene (gls) was cloned and overexpressed in Ganoderma lingzhi. The content of intra-cellular polysaccharides (IPS) in G. lingzhi overexpressing gls was 22.36 mg/100 mg dry weight (DW), 19 % higher than those in the wild-type (WT) strain. Overexpression of gls did not affect the expression of the phos-phoglucomutase gene and the UDP-glucose pyrophosphorylase gene (ugp) in the polysaccharide biosynthesis. The gls and ugp were then simultaneously overexpressed in G. lingzhi for the first time. The combined over-expression of these two genes increased the IPS content and exopolysaccharides (EPS) production to a greater extent than the overexpression of gls independently. The maximum IPS content of the overexpressed strain was 24.61 mg/100 mg, and the maximum EPS production was 1.55 g/L, 1.31-and 1.50-fold higher than that in the WT strain, respectively. Moreover, the major EPS fractions from the overexpression strain contained more glucose (86.7 % and 72.5 %) than those from the WT strain (78.2 % and 62.9 %). Furthermore, the major fraction G+U-0.1 from the overexpression strain exhibited stronger antioxidant and anti-senescence activities than the WT-0.1 fraction from the WT strain. These findings will aid in the hyperproduction and application of Ganoderma polysaccharides and facilitate our understanding of mushroom polysaccharide biosynthesis.

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