Increased polysaccharide production and biosynthetic gene expressions in a submerged culture of Ganoderma lucidum by the overexpression of the homologous α-phosphoglucomutase gene

Xu, Junwei; Ji, Senlin; Li, Huanjun; Zhou, Jinhua; Duan, Yan-Qing; Dang, Li-Zhi; Mo, Ming-He

doi:10.1007/s00449-014-1279-1

Cited by 71 publications

(54 citation statements)

References 39 publications

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“…The above results suggest that the observed increase in bioflocculant secretion resulted from enhanced expression of several key genes, which promoted metabolite synthesis in the polysaccharide metabolic pathway (Xu et al, 2015). Contrary to what we expected, however, the polysaccharide bioflocculant yield and flocculating activity of the recombinant strains pHY300PLK-mnaA1 and pHY300PLK-mnaA2 both decreased.…”

Section: Effect Of Enhanced Expression Of Differential Gene On Eps Yieldcontrasting

confidence: 67%

Identification of key genes involved in polysaccharide bioflocculant synthesis in Bacillus licheniformis

Chen

Liu

et al. 2016

Biotech & Bioengineering

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The present study reports the sequenced genome of Bacillus licheniformis CGMCC 2876, which is composed of a 4,284,461 bp chromosome that contains 4,188 protein-coding genes, 72 tRNA genes, and 21 rRNA genes. Additional analysis revealed an eps gene cluster with 16 open reading frames. Conserved Domains Database analysis combined with qPCR experiments indicated that all genes in this cluster were involved in polysaccharide bioflocculant synthesis. Phosphoglucomutase and UDP-glucose pyrophosphorylase were supposed to be key enzymes in polysaccharide secretion in B. licheniformis. A biosynthesis pathway for the production of polysaccharide bioflocculant involving the integration of individual genes was proposed based on functional analysis. Overexpression of epsDEF from the eps gene cluster in B. licheniformis CGMCC 2876 increased the flocculating activity of the recombinant strain by 90% compared to the original strain. Similarly, the crude yield of polysaccharide bioflocculant was enhanced by 27.8%. Overexpression of the UDP-glucose pyrophosphorylase gene not only increased the flocculating activity by 71% but also increased bioflocculant yield by 13.3%. Independent of UDP-N-acetyl-D-mannosamine dehydrogenase gene, flocculating activity, and polysaccharide yield were negatively impacted by overexpression of the UDP-N-acetylglucosamine 2-epimerase gene. Overall, epsDEF and gtaB2 were identified as key genes for polysaccharide bioflocculant synthesis in B. licheniformis. These results will be useful for further engineering of B. licheniformis for industrial bioflocculant production. Biotechnol. Bioeng. 2017;114: 645-655. © 2016 Wiley Periodicals, Inc.

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Section: Effect Of Enhanced Expression Of Differential Gene On Eps Yieldcontrasting

confidence: 67%

Identification of key genes involved in polysaccharide bioflocculant synthesis in Bacillus licheniformis

Chen

Liu

et al. 2016

Biotech & Bioengineering

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“…The preparation of G. lucidum protoplasts and PEG‐mediated genetic transformation were conducted as previously described (Xu et al , , ; Yu et al , ). Transformants were screened using carboxin (2 mg l −1 ) or phosphinothricin (150 mg l −1 ) on a CYM regeneration plate.…”

Section: Methodsmentioning

confidence: 99%

Dual sgRNA‐directed gene deletion in basidiomycete Ganoderma lucidum using the CRISPR/Cas9 system

Liu

Sun

You

et al. 2020

Microbial Biotechnology

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Ganoderma lucidum is an important medicinal mushroom in traditional Chinese medicine. However, the lack of adequate genetic tools has hindered molecular genetic research in and the genetic modification of this species. Here, we report that the presence of an intron is necessary for the efficient expression of the heterologous phosphinothricin‐resistance and green fluorescent protein genes in G. lucidum. Moreover, we improved the CRISPR/Cas9‐mediated gene disruption frequency in G. lucidum by adding an intron upstream of the Cas9 gene. Our results showed that the disruption frequency of the orotidine 5’‐monophosphate decarboxylase gene (ura3) in transformants containing the glyceraldehyde‐3‐phosphate dehydrogenase gene intron in the Cas9 plasmid is 14–18 in 107 protoplasts, which is 10.6 times higher than that in transformants without any intron sequence. Furthermore, genomic fragment deletions in the ura3 and GL17624 genes were achieved via a dual sgRNA‐directed CRISPR/Cas9 system in G. lucidum. We achieved a ura3 deletion frequency of 36.7% in G. lucidum. The developed method provides a powerful platform to generate gene deletion mutants and will facilitate functional genomic studies in G. lucidum.

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“…This has been demonstrated as a promising approach for the production of various compounds including polysaccharides (Fang and Zhong, 2002, Wagner et al., 2003, Habijanic et al., 2015). Genetically engineering strains to increase the production of ganodermic acids in submerged cultures has also enjoyed some success (Zhou et al., 2014, Xu et al., 2015, Li et al., 2016). As well as being a faster approach, the use of submerged cultures allows improved control over various fermentation conditions such as nutrient availability, pH and oxygen, which allows optimisation and greater reproducibility.…”

Section: Medicinal Basidiomycetesmentioning

confidence: 99%

The good, the bad and the tasty: The many roles of mushrooms

Mattos-Shipley¹,

Ford²,

Alberti³

et al. 2016

Studies in Mycology

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Fungi are often inconspicuous in nature and this means it is all too easy to overlook their importance. Often referred to as the “Forgotten Kingdom”, fungi are key components of life on this planet. The phylum Basidiomycota, considered to contain the most complex and evolutionarily advanced members of this Kingdom, includes some of the most iconic fungal species such as the gilled mushrooms, puffballs and bracket fungi. Basidiomycetes inhabit a wide range of ecological niches, carrying out vital ecosystem roles, particularly in carbon cycling and as symbiotic partners with a range of other organisms. Specifically in the context of human use, the basidiomycetes are a highly valuable food source and are increasingly medicinally important. In this review, seven main categories, or ‘roles’, for basidiomycetes have been suggested by the authors: as model species, edible species, toxic species, medicinal basidiomycetes, symbionts, decomposers and pathogens, and two species have been chosen as representatives of each category. Although this is in no way an exhaustive discussion of the importance of basidiomycetes, this review aims to give a broad overview of the importance of these organisms, exploring the various ways they can be exploited to the benefit of human society.

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Increased polysaccharide production and biosynthetic gene expressions in a submerged culture of Ganoderma lucidum by the overexpression of the homologous α-phosphoglucomutase gene

Cited by 71 publications

References 39 publications

Identification of key genes involved in polysaccharide bioflocculant synthesis in Bacillus licheniformis

Identification of key genes involved in polysaccharide bioflocculant synthesis in Bacillus licheniformis

Dual sgRNA‐directed gene deletion in basidiomycete Ganoderma lucidum using the CRISPR/Cas9 system

The good, the bad and the tasty: The many roles of mushrooms

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