Overexpression of phosphomannomutase increases the production and bioactivities of Ganoderma exopolysaccharides

Zhao, Lei; Cao, Yubiao; Luo, Qin; Xu, Yong-Liang; Li, Na; Wang, Chengxiao; Xu, Junwei

doi:10.1016/j.carbpol.2022.119828

Cited by 12 publications

(6 citation statements)

References 34 publications

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“…plantarum to regulate the intestinal barrier is partly performed by genes involved in CPS synthesis based on the comparative genomic analysis. Zhao et al [40] confirmed the overexpression of phosphomannomutase the increase the production of Ganoderma EPSs and improve its bioactivities. Admittedly, the genes encoding proteins associated with EPS biosynthesis are essential for its production and potential function.…”

Section: Relationship Between Eps Biosynthesis and Immunomodulatory E...mentioning

confidence: 96%

Molecular mechanism of exopolysaccharide biosynthesis regulated by wze and wzd genes on the immunomodulatory effects of probiotic Lacticaseibacillus paracasei S-NB

Xiao,

Yang,

Zhang

et al. 2024

Food Science and Human Wellness

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The surface macromolecules of probiotic bacteria play crucial roles in modulating immune responses in the host. Exopolysaccharide (EPS) from lactic acid bacteria (LAB) have been widely reported to exhibit immunomodulatory activity. In this study, the EPS biosynthesis gene cluster of Lacticaseibacillus paracasei S-NB was analyzed and a deletion mutant S-NB△7576 (two genes S-NB_2175/wze and S-NB_2176/wzd were responsible for the chain length determination and export of EPS) was successfully constructed, resulting a 40.02% decrease in the production of EPS. The deletion of wze and wzd had little effect on the monosaccharide composition and major groups of the two EPS fractions (BEPS1 and BEPS2). Both BEPS1 and BEPS2 could inhibit the transcriptional level of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA in RAW 264.7 cells induced by lipopolysaccharide (LPS), and enhance host immune tolerance via suppressing NF-κB and MAPK signaling. Notably, the S-NB△7576 mutant supplied with the BEPS1/BEPS2 exhibited more significant inhibition of cytokines production and the phosphorylation of p65 and c-Jun N-terminal kinase (JNK) in LPS-stimulated cells compared with the S-NB△7576 mutant alone. Our study provided the immunomodulatory effect of BEPS1 and BEPS2 from L. paracasei S-NB, in which the wze and wzd genes associated with EPS biosynthesis may play an important role.

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Section: Relationship Between Eps Biosynthesis and Immunomodulatory E...mentioning

confidence: 96%

Molecular mechanism of exopolysaccharide biosynthesis regulated by wze and wzd genes on the immunomodulatory effects of probiotic Lacticaseibacillus paracasei S-NB

Xiao,

Yang,

Zhang

et al. 2024

Food Science and Human Wellness

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“…The gene-specific primers (Table S1) were designed with Primer Premier 6.0 software (Premier Biosoft International, Palo Alto, CA, USA). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was chosen as an endogenous reference gene to normalize target gene expression (Lu et al, 2018;Zhao et al, 2022). qRT-PCR was performed using TB Green® Premix Ex Taq (TAKARA, Dalian, China) on Light cycle 96 Real-Time PCR System (Roche, Switzerland).…”

Section: Quantitative Real-time Pcr (Qrt-pcr) Analysismentioning

confidence: 99%

“…There are several adenosine derivatives in G. lucidum . Adenosine-rich extract of G. lucidm is reported to possess adequate lipid-lowering and anti-atherosclerotic effects (Li et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Overexpression of purine nucleoside phosphorylase increases the adenosine content in Ganoderma lucidum

Liu

Zhu

Xiao

et al. 2022

Preprint

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Adenosine has been getting increasing attention due to its positive role in immunomodulation, anti-inflammation, and anti-cancer, etc.. The low production of Ganoderma adenosine is a bottleneck for clinical trials and commercial applications. Regulating the expression of key biosynthetic gene is an optimized way to increase the adenosine production in submerged culture of Ganoderma lucidum. In this study, we correlated the expression of adenosine synthase genes (including GlATIC, GlPNP, GlADK) with the adenosine content. The results showed that GlPNP was positively correlated with Ganoderma adenosine contents. Then the key biosynthetic gene GlPNP was cloned, characterized and overexpressed in G. lucidum. The cDNA of GlPNP gene was 969-bp in length, with a predicted molecular weight of 34.6 kDa. The GlPNP displayed a trimeric quaternary structure. The transcript levels of GlPNP overexpression transformants were approximately 2.9-3.9-fold higher than those of the WT strains on day 4, while the adenosine contents were increased by 78% and 63%, respectively, by compared with vector-containing strain. The GlPNP overexpression strains showed decreased colony growth and reduced biomass. In conclusion, GlPNP gene overexpression is an effective strategy to improve the production of adenosine. This study is the first report about the manipulation of adenosine biosynthesis in medicinal fungi.

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“…The gene-speci c primers (Table S1) were designed with Primer Premier 6.0 software (Premier Biosoft International, Palo Alto, CA, USA). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was chosen as an endogenous reference gene to normalize target gene expression [29,30]. qRT-PCR was performed using TB Green® Premix Ex Taq™ (TAKARA, Dalian, China) on Light cycle 96 Real-Time PCR System (Roche, Switzerland).…”

Section: Nucleic Acid Isolationmentioning

confidence: 99%

Overexpression of purine nucleoside phosphorylase increases the adenosine content in Ganoderma lucidum

Zhu

Xiao

Zhou

et al. 2022

Preprint

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Background: Adenosine has been getting increasing attention due to its positive role in immunomodulation, anti-inflammation, and anti-cancer, etc.. The low production of Ganoderma adenosine is a bottleneck for clinical trials and commercial applications. Regulating the expression of key adenosine biosynthetic gene is an optimized way to increase the production of adenosine in submerged culture of Ganoderma lucidum. Results: In this study, we correlated the expression of adenosine synthase genes (including GlATIC, GlPNP, GlADK) with the adenosine content in mycelium at different fermentation time points. The results showed that GlPNP was positively correlated with Ganoderma adenosine contents. Then the key biosynthetic gene GlPNP was cloned, characterized and overexpressed in G. lucidum. The cDNA of GlPNP gene was 969-bp in length, with a predicted molecular weight of 34.6 kDa and PI of 5.89. The GlPNP displayed a trimeric quaternary structure by theoretically modelling with SWISS-MODEL. The transcript levels of GlPNP overexpression transformants (namely OE::GlPNP-5 and OE::GlPNP-7) were approximately 2.9-3.9-fold higher than those of the WT strains on day 4, while the adenosine contents were increased by 78% and 63%, respectively, by compared with vector-containing strain. In addition, the GlPNP overexpression strains showed decreased colony growth and reduced biomass in submerged cultivation. Conclusions: GlPNP gene overexpression is an effective strategy to improve the production of adenosine in G. lucidum. This study is the first report about the manipulation of adenosine biosynthesis in medicinal fungi.

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Overexpression of phosphomannomutase increases the production and bioactivities of Ganoderma exopolysaccharides

Cited by 12 publications

References 34 publications

Molecular mechanism of exopolysaccharide biosynthesis regulated by wze and wzd genes on the immunomodulatory effects of probiotic Lacticaseibacillus paracasei S-NB

Molecular mechanism of exopolysaccharide biosynthesis regulated by wze and wzd genes on the immunomodulatory effects of probiotic Lacticaseibacillus paracasei S-NB

Overexpression of purine nucleoside phosphorylase increases the adenosine content in Ganoderma lucidum

Overexpression of purine nucleoside phosphorylase increases the adenosine content in Ganoderma lucidum

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