Improved polysaccharide production in a submerged culture of Ganoderma lucidum by the heterologous expression of Vitreoscilla hemoglobin gene

Li, Huanjun; Zhang, De-Huai; Yue, Tong-Hui; Lei, Jiang; Yu, Xuya; Zhao, Peng; Li, Tao; Xu, Junwei

doi:10.1016/j.jbiotec.2015.11.011

Cited by 44 publications

(34 citation statements)

References 41 publications

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“…However, the VHb expression did not improved the growth (dry mycelia weight) of the transformants in glucose-containing medium, nor did it increase the biomass of TU6-vgb + in the cellulose-containing medium. These results were consistent with the studies in E. coli (Geckil et al 2001 ), Serratia marcescens (Wei et al 1998 ) and Ganoderma lucidum (Li et al 2016 ) that the VHb-expressing strains did not grow better than the strains without VHb, but contrary to the study in A. sojae (Mora-Lugo et al 2015 ) and P. lilacinus (Zhang et al 2014 ). It was perhaps that the effect of VHb on the biomass may vary in different organisms.…”

Section: Discussionsupporting

confidence: 87%

Improving cellulase production in submerged fermentation by the expression of a Vitreoscilla hemoglobin in Trichoderma reesei

et al. 2017

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Trichoderma reesei is well known as an industrial workhorse fungus in cellulase production. The low dissolved oxygen supply in the highly viscous medium of T. reesei remains a major bottleneck that hampers growth and cellulase production in submerged fermentation. Vitreoscilla hemoglobin (VHb) has been demonstrated to improve metabolism and protein production in different heterologous hosts under hypoxic conditions, but the use of VHb in T. reesei remains uninvestigated. This study examines the effect of VHb in improving T. reesei performance in submerged fermentation. The VHb gene (vgb)-expressing cassette was successfully transformed into the TU-6 strain, integrated into the genome of T. reesei, and functionally expressed with biological activity, which was confirmed by carbon monoxide difference analysis. Compared to the parent strain, the expression of VHb increased the glucose consumption rate of the transformant. Moreover, in cellulase-inducing medium total protein secretion of the VHb expressing strain was 2.2-fold of the parental strain and the filter paper cellulase activity was increased by 58% under oxygen-limiting conditions. In summary, our results demonstrate that VHb has beneficial effects on improving total protein secretion and cellulase activity of T. reesei in submerged fermentation.Electronic supplementary materialThe online version of this article (10.1186/s13568-017-0507-x) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 87%

Improving cellulase production in submerged fermentation by the expression of a Vitreoscilla hemoglobin in Trichoderma reesei

et al. 2017

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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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“…Genetic transformation systems, including Agrobacterium tumefacien ‐mediated and polyethylene glycol (PEG)‐mediated transformations, have been developed in G. lucidum (Shi et al , ; Xu et al , ; Xu and Zhong, ) and have been successfully used to enhance the production of bioactive compounds, such as ganoderic acids and polysaccharides (Zhou et al , ; Li et al , ; Zhang et al , , ; Fei et al , ; Xu et al , ). Gene silencing systems have also been used for the partial suppression of genes of interest in G. lucidum (Mu et al , ; Liu et al , ; Zhu et al , ).…”

Section: Introductionmentioning

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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Improved polysaccharide production in a submerged culture of Ganoderma lucidum by the heterologous expression of Vitreoscilla hemoglobin gene

Cited by 44 publications

References 41 publications

Improving cellulase production in submerged fermentation by the expression of a Vitreoscilla hemoglobin in Trichoderma reesei

Improving cellulase production in submerged fermentation by the expression of a Vitreoscilla hemoglobin in Trichoderma reesei

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

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

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