Molecular Cloning, Characterisation, and Heterologous Expression of Farnesyl Diphosphate Synthase from Sanghuangporus baumii

Biotechnology & Biotechnological Equipment

2020

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Sanghuangporus baumii (Pil at) L.W. Zhou & Y.C. Dai, a popular medicinal fungus, has been used to treat several diseases by its triterpenoids bioactive ingredient for many years. However, the triterpenoids content of S. baumii is low and their biosynthesis mechanisms are not clearly understood. In order to reveal the regulation mechanism of triterpenoids biosynthesis in S. baumii, the cDNA encoding isopentenyl diphosphate isomerase (IDI) involved in triterpenoids biosynthesis was cloned and named as SbIDI (GenBank number MK955885). The open reading frame of the SbIDI gene cDNA comprised 792 bp and encoded a polypeptide of 263 amino acids with a predicted protein molecular weight of 29.80 kDa. The transcript level of the SbIDI gene and triterpenoids content of S. baumii were detected at different developmental stages. The results showed that the transcript level of the SbIDI gene and triterpenoids content had almost the same trend of change, increased first and then decreased dynamically. The highest transcript level of the SbIDI gene occurred earlier than the highest triterpenoids content, indicating that the SbIDI gene may play an important role in triterpenoids biosynthesis in S. baumii. Moreover, the SbIDI gene cDNA was successfully expressed in Escherichia coli BL21 (DE3), and the protein bands were consistent with the prediction.The results will lay a foundation for further study of the function of the SbIDI gene.

Section: Transcript Level Of Sbidi Gene At Different Developmental Stsupporting

confidence: 85%

Cloning, molecular properties and differential expression analysis of the isopentenyl diphosphate isomerase gene in Sanghuangporus baumii

Liu

Sun

Biotechnology & Biotechnological Equipment

2020

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“…These regulatory elements control the expression of genes in the MVA pathway in response to various hormones and stressors. In our previous studies, similar cis-regulatory elements were also found in the homologous promoters of the key enzyme genes of S. baumii's MVA pathway, acetyl-CoA C-acetyltransferase synthase gene [29] and farnesyl pyrophosphate synthase gene [30]. Moreover, under the stimulation of MeJA, the content of triterpenoids in the mycelia of S. baumii increased by 4 times [31], which may be related to the MeJA-responsiveness elements in the promoters.…”

Section: Sbpmk Expression and Triterpenoids Content During Fermentationsupporting

confidence: 69%

Cloning and characterization of a phosphomevalonate kinase gene from Sanghuangporus baumii

Liu

Biotechnology & Biotechnological Equipment

et al. 2021

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Sanghuangporus baumii, a macrofungus belonging to the Hymenochaetaceae family, has been widely used as a traditional medicine in Asia for centuries. Triterpenoids in S. baumii contribute to the pharmacological activities, however, their content is low in the fruiting body. Phosphomevalonate kinase (PMK) is an important rate-limiting enzyme in the triterpenoids synthesis pathway. In this study, a PMK gene (SbPMK) was cloned and analyzed. Sequence analysis showed that SbPMK has six exons and five introns. The 1,488 bp open reading frame encodes a 489 amino acid polypeptide with a predicted protein molecular weight of 52.47 kDa. The amino acid sequence of SbPMK shares highly similarity with that of PMK sequences from other species, especially in three functional motifs (MI, MII and MIII). The promoter of SbPMK contains typical cis-acting regulatory elements involved in MeJA responsiveness, light responsiveness, anaerobic induction, etc. Furthermore, recombinant SbPMK was overexpressed in Escherichia coli BL21, with a satisfactory solubility. During the fermentation culture of S. baumii, the biomass increased with fermentation and peaked on day 18 (4.80 g/L). The SbPMK expression and the triterpenoids content was peaked on day 6, and the triterpenoids content stabilized at 20.8 mg/g on day 18.

“…For thousands of years, many basidiomycetes, such as Lentinus edodes , Auricularia auricula and Hericium erinaceus , have been used as food (Lei et al 2019 ; Zhao et al 2020 ; Miao et al 2020 ; Hu et al 2020 ). Additionally, certain fungi, such as Ganoderma lucidum and Poria cocos (Lin et al 2020 ; Wang et al 2020a , b , c ), have been used as medicinal plants. Basidiomycetes, particularly their sporocarps, contain abundant natural products (Tian et al 2020 ).…”

Section: Introductionmentioning

confidence: 99%

“…Compared with the 9-day hyphal control, the expression in the 11- and 13-day hyphal groups was upregulated by 49.3 times and 125.4 times, respectively. Through analysis, the triterpene content of S. baumii was found to be significantly correlated with the expression level of FPS at different developmental stages (Wang et al 2020a , b , c ). The results of this experiment showed that FPS genes are involved in regulating the biosynthesis of triterpenoids.…”

Section: Introductionmentioning

confidence: 99%

“…G. lucidum is the fungus with the most triterpenoids, and more than 250 triterpenoid compounds have been identified from this species. However, compared with plants, we know relatively little regarding the biosynthesis of triterpenoid compounds in basidiomycetes (Wang et al 2020a , b , c ). It has been determined that the primary terpene component in G. lucidum is ganoderic acid (GA) (Shao et al 2020 ; Liang et al 2019 ), and more than 170 types of GA have been isolated from G. lucidum (Wang et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Biosynthesis and regulation of terpenoids from basidiomycetes: exploration of new research

et al. 2021

Basidiomycetes, also known as club fungi, consist of a specific group of fungi. Basidiomycetes produce a large number of secondary metabolites, of which sesquiterpenoids, diterpenoids and triterpenoids are the primary components. However, these terpenoids tend to be present in low amounts, which makes it difficult to meet application requirements. Terpenoid biosynthesis improves the quantity of these secondary metabolites. However, current understanding of the biosynthetic mechanism of terpenoids in basidiomycetes is insufficient. Therefore, this article reviews the latest research on the biosynthesis of terpenoids in basidiomycetes and summarizes the CYP450 involved in the biosynthesis of terpenoids in basidiomycetes. We also propose opportunities and challenges for chassis microbial heterologous production of terpenoids in basidiomycetes and provide a reference basis for the better development of basidiomycete engineering.