Genetic engineering ofGanoderma lucidumfor the efficient production of ganoderic acids

Abstract

Ganoderma lucidum is a well-known traditional medicinal mushroom that produces ganoderic acids with numerous interesting bioactivities. Genetic engineering is an efficient approach to improve ganoderic acid biosynthesis. However, reliable genetic transformation methods and appropriate genetic manipulation strategies remain underdeveloped and thus should be enhanced. We previously established a homologous genetic transformation method for G. lucidum; we also applied the established method to perform the deregulated overexpression of a homologous 3-hydroxy-3-methyl-glutaryl coenzyme A reductase gene in G. lucidum. Engineered strains accumulated more ganoderic acids than wild-type strains. In this report, the genetic transformation systems of G. lucidum are described; current trends are also presented to improve ganoderic acid production through the genetic manipulation of G. lucidum.

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

“…The G. lucidum FPS gene was amplified by genomic PCR with the primers FPS- Nhe I-F and FPS- Sma I-R. To overexpress the FPS gene, the plasmid pJW-EXP-FPS (Additional file 1: Fig. S1) was constructed and introduced into G. lucidum using the reported genetic transformation method [10, 14, 41]. The expression of the FPS gene was driven by the glyceraldehyde-3-phosphate dehydrogenase gene promoter in the pJW-EXP-FPS plasmid.…”

“…GAs are biosynthesized via the mevalonate/isoprenoid (MVA) pathway from acetyl-coenzyme A to produce lanosterol in Ganoderma species (Fig. 1) [8–10]. Further biosynthetic steps to form GAs probably include a series of complex oxidation, reduction, hydroxylation, and acylation reactions [8, 11].…”

“…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.…”

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 , ).…”

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

“…Due to their important pharmacological activities, many strategies have been used to increase the production of GAs during mycelia fermentation. Progress has been achieved in the last several decades in enhancing GA production in G. lucidum by the manipulation of fermentation strategies (Xu et al, 2010a;Upadhyay et al, 2014;Wang et al, 2016), addition of metal ions and elicitors You et al, 2013You et al, , 2017Zhang et al, 2014;Gu et al, 2018) and genetic engineering Xu and Zhong, 2015;Li et al, 2016a,b,c;Zhang et al, 2017a,b). However, further improvement of GA production is necessary to meet the demand of increasing preclinical studies and commercial applications.…”