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)

Ji, Senlin; Li, Rui; Ren, Meng-Fei; Li, Huanjun; Xu, Junwei

doi:10.1615/intjmedmushrooms.v17.i5.30

Cited by 36 publications

(29 citation statements)

References 15 publications

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“…The generation of nucleotide sugar precursor is an important and basic part in the process of CP biosynthesis. As researchers have discovered in their work, the overexpression of genes involved in the polysaccharide biosynthetic pathway can increase the production of Ganoderma lucidum polysaccharides [11,12]. Knocking out genes in the branch pathway and controlling the metabolic pathways direction of products can also affect the yield of polysaccharides too.…”

Section: Introductionmentioning

confidence: 99%

Homologous Overexpression of Genes in Cordyceps militaris Improves the Production of Polysaccharide

Yang

Wang

Chen

et al. 2020

Preprint

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Background Cordyceps polysaccharides have been used around the globe for its bioactivity for millennia. However, the study and medicinal of Cordyceps militaris polysaccharides has been hampered by the low in natural abundances. Recently, the genetic engineered C. militaris developed for production of exopolysaccharides (EPS) had received extensive attention. ResultsIn this study, based on the biosynthetic pathway and metabolization mechanism of exopolysaccharides, the crucial biosynthetic genes of Cordyceps polysaccharides were introduced by Agrobacterium transformation to provide a high flux of EPS. 21 mutants of C. militaris were identified through antibiotic screening and DNA sequencing. The maximum yield of EPS produced by mutant CM-pgm-H was 4.63 ± 0.23 g/L, while the yield of wild-type strain was 3.43 ± 0.26 g/L. And the data obtained in the present study indicated that the yield of EPS produced by the engineered strain treated with co-overexpression of phosphoglucomutase and UDP-glucose 6-dehydrogenase genes achieved 6.11 ± 0.21 g/L, which was increased by 78.13% compared with the wild-type strain.Conclusions CM-pgm-H obtained the highest EPS content than that of mutants glucokinase, UDPglucose pyrophosphorylase, UDP-glucose 6-dehydrogenase. It indicated that the content of protein phosphoglucomutase was the most critical influencing factor on the CP production in C.militaris.Furthermore, the EPS production of CM-ugdh-pgm-M was significantly improved 1.78-fold by cooverexpression. It anticipated that our engineering strategies will play an important role in the development of C. militaris for sustainable production of Cordyceps polysaccharides. BackgroundCordyceps militaris (L.:Fr.) Fr. is an entomogenous fungus belonging to Ascomycetes, Hypocreales, Cordycipitaceae, Cordyceps (L.:Fr.) Link [1], and has received extensive research and attention because of its potential medicinal properties. The production of natural Cordyceps decreased year by year, while its market demand is still great, so more and more research groups are focusing on submerge fermentation of Cordyceps mycelia. Nowadays, many biologically active substances have been isolated from C. militaris, such as Cordyceps polysaccharides (CP), Cordycepic acid, cordycepin, mannitol, alkaloids, vitamins B1, B2, and minerals. It was reported that CP has a good protection of

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Section: Introductionmentioning

confidence: 99%

Homologous Overexpression of Genes in Cordyceps militaris Improves the Production of Polysaccharide

Yang

Wang

Chen

et al. 2020

Preprint

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“…Xu et al reported that overexpressing the alpha phosphoglucomutase gene could increase the yield of Ganoderma polysaccharides [ 26 ]. Ji et al found that overexpression of the UDP glucose pyrophosphorylase (UGP) gene would increase the production of polysaccharides [ 27 ].…”

Section: Introductionmentioning

confidence: 99%

Improved production of polysaccharides in Ganoderma lingzhi mycelia by plasma mutagenesis and rapid screening of mutated strains through infrared spectroscopy

Zhang

et al. 2018

PLoS ONE

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As a traditional Chinese medicine, Ganoderma lingzhi has attracted increasing attention for both scientific research and medical application. In this work, in order to improve the production of polysaccharides from an original wide-type (WT) strain (named "RWY-0") of Ganoderma lingzhi, we applied atmospheric-pressure dielectric barrier discharge (DBD) nonthermal plasma to the protoplasts of RWY-0 for mutagenesis treatment. Through a randomly amplified polymorphic DNA (RAPD) assay, at least 10 mutagenic strains were confirmed. They also showed different mycelium characteristics in terms of shape, color, size and biomass in liquid fermentation. The mutant strains were examined by infrared spectroscopy, and based on the established near-infrared (NIR) quantification model, the polysaccharide contents in these mutants were quantitatively evaluated. As a result, we found that the Ganoderma polysaccharide contents in some of the mutant strains were significantly changed compared with that of the original WT strain. The polysaccharide content of RWY-1 G. lingzhi was considerably higher than that of the WT strain, with an increase of 25.6%. Thus, this preliminary work demonstrates the extension of the plasma mutagenesis application in acquiring polysaccharide-enhanced Ganoderma lingzhi mutants and shows the usefulness of NIR spectroscopy in the rapid screening of mutagenic strains for other important ingredients.

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“…Several months are usually needed to cultivate the G. lucidum fruiting body, and product quality is difficult to control using standard methods . Many researchers have focused on the culture medium, fermentation modes to accelerate the mycelial growth and fermentation to activate the mycelium to produce polysaccharides . These polysaccharides or extracts of polysaccharides have been used in traditional medicine as antitumor and immunomodulating agents .…”

Section: Introductionmentioning

confidence: 99%

Enhanced triterpene acid production by Ganoderma lucidum using a feeding stimulus integrated with a two‐stage pH‐control strategy

Wang

Yang

Liu

2016

J of Chemical Tech & Biotech

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BACKGROUND Triterpenes (especially triterpene acids) are some of the major physiologically active components of Ganoderma lucidum. However, submerged fermentation techniques for triterpenes have not been optimized for commercial use, and the total triterpene yield is still very low. The aim of this work was to develop a two‐stage pH‐control strategy integrated with the use of an ether extract from the medicinal insect, Catharsius molossus (EECM), as a stimulus for enhanced G. lucidum triterpene acid production. RESULTS Triterpene acid production of 302.76 mg L−1 was obtained in G. lucidum cultured with the EECM; this production was higher by 60% than that of the control cultures not fed the EECM. Based on the EECM feeding method, a new two‐stage pH‐control strategy was integrated, resulting in triterpene acid yields of 361.47 mg L−1. Subsequent studies optimized the interaction between EECM feeding and two‐stage pH‐control strategies, which resulted in an optimal EECM concentration of 185.89 mg L−1 and optimal pH values of 5.07 in the first 72 h incubation and 4.37 after 72 h, with a maximum triterpene acid concentration of 449.37 mg L−1. This represented a 137% increase compared with that of the control cultures. Furthermore, when a 150 L fermenter was used under these conditions, a triterpene acid concentration of 431.83 mg L−1 was achieved. CONCLUSIONS The enhanced production of triterpene acid in submerged cultures of G. lucidum was achieved using an EECM feeding stimulus integrated with a two‐stage pH‐control strategy at large scale. © 2016 Society of Chemical Industry

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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)

Cited by 36 publications

References 15 publications

Homologous Overexpression of Genes in Cordyceps militaris Improves the Production of Polysaccharide

Homologous Overexpression of Genes in Cordyceps militaris Improves the Production of Polysaccharide

Improved production of polysaccharides in Ganoderma lingzhi mycelia by plasma mutagenesis and rapid screening of mutated strains through infrared spectroscopy

Enhanced triterpene acid production by Ganoderma lucidum using a feeding stimulus integrated with a two‐stage pH‐control strategy

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