Enhanced production of individual ganoderic acids by integrating Vitreoscilla haemoglobin expression and calcium ion induction in liquid static cultures of Ganoderma lingzhi

Abstract

Summary Ganoderic acids produced by Ganoderma exhibit anticancer and antimetastatic activities. A novel approach by combining Vitreoscilla haemoglobin (VHb) expression and calcium ion induction was developed to enhance ganoderic acid (GA) production in liquid static cultures of G. lingzhi. The maximum contents of GA‐O, GA‐S and GA‐Me were 1451.33 ± 67.50, 1431.23 ± 79.74 and 1283.81 ± 85.13 μg per 100 mg cell weight, respectively under the integrated approach, which are the highest contents as ever reported in Ganoderma. The contents of squalene and lanosterol were increased by 2.0‐ and 3.0‐fold in this case compared with those in the control. The transcription levels of 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase, farnesyl‐diphosphate synthase, squalene synthase and cytochrome P450 CYP5150L8 were upregulated by 2.56‐, 3.31‐, 2.59‐ and 6.12‐fold respectively. Additionally, the expression of VHb improved the ratio of type I to type II GA in liquid static cultivation of G. lingzhi. The transcription levels of cyp512a2, cyp512v2 and cyp512a13, candidate cytochrome P450 genes involved in oxidative modification of the lanostane skeleton in GA biosynthesis, were also increased by 2.28‐, 2.65‐ and 3.54‐fold in the VHb‐expressing strain respectively. Our results illustrated that the approach described here efficiently improved GA production in G. lingzhi fermentation.

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

“…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 , ).…”

Increased production of ganoderic acids by overexpression of homologous farnesyl diphosphate synthase and kinetic modeling of ganoderic acid production in Ganoderma lucidum

“…Compared to fruiting body cultures, a submerged culture is a promising alternative for the production of triterpenoids, as it is easy to control the product quality and is cost effective [21, 22]. Many attempts, such as by manipulating fermentation conditions [6, 23], developing bioprocessing and elicitor strategies [24–29], and metabolic engineering [10, 14, 15, 17, 18, 30, 31], have been conducted to increase the production of GAs by fermentation of mycelia.…”

Transcriptome dynamics and metabolite analysis revealed the candidate genes and regulatory mechanism of ganoderic acid biosynthesis during liquid superficial‐static culture of Ganoderma lucidum

“…However, only AACT‐1 gene expressed differently on fifth and sixth days which was upregulated in LSSC that meant it may be a regulated gene for total triterpenoid biosynthesis. HMGR was also recognized as a key point for GA biosynthesis, because the transcription levels of HMGR were increased in a GA high‐yield Ganoderma sp (Xu et al ., 2010a, 2019), and ganoderic triterpenoid content was increased in the HMGR overexpressed strain (Xu et al ., 2012). In this study, HMGR gene was upregulated on days 5 and 6; however, it was downregulated in the next two days.…”

“…(2020). The primers were designed using NCBI Primer Blast and referring to previous research (Table S1) (Xu et al ., 2010a, 2019). For each gene analysed by RT‐qPCR, three biological replicates were performed.…”

“…Recently, Xu et al . (2019) reported that CYP5150L8 , CYP512A2 , CYP512V2 and CYP512A13 genes were associated with the oxidative modification of the lanostane skeleton in GA biosynthesis and that the transcription levels of these genes were increased in a GA high‐yielding strain.…”