Abstract
Ganoderma lucidum, a famous medicinal mushroom used worldwide, is a rich source of triterpenoids which, together with polysaccharides, are believed to be the main effective constituents of G. lucidum. With the increase of market demand, the wild resource is facing serious limitations, and the quality of cultivated fruiting bodies can be seriously affected by the availability of wood resources and by cultivation management practices. In the present study, we aimed to develop an alternative way to produce useful triterpenoids from G. lucidum. We cultured the strain using a two-stage liquid culture strategy and investigated the effects of nitrogen limitation, carbon supply, static culture volume and air supply in the static culture stage on the accumulation of five triterpenoids (GA-P, GA-Q, GA-T, GA-S, GA-R). Our results showed that, under optimized condition, the total yield of the five triterpenoids reached 963 mg/L (as determined by HPLC). Among the five triterpenoids, GA-T accounted for about 75% of the total yield. Besides, a bioreactor suitable for fungal liquid static culture with a 10 L extensible plastic bag shaped culture unit was designed and in which the maximum total yield of the five GAs reached 856.8 mg/L, and the GAs content reached 5.99%. Our results demonstrate the potential of industrial application of G. lucidum culture for the production of triterpenoids, especially GA-T. Air supply significantly improved the accumulation of triterpenoids, and this will provide important clues to understand why more triterpenoids are produced in the mycelia mat under static liquid culture conditions.
Highlights
Higher fungi, especially Ascomycetes, Deuteromycetes, and Basidiomycetes [1], have been attracting more and more attention in recent decades for their abundant bioactive secondary metabolites and their potential to be developed as drugs and health foods
For the RP-High performance liquid chromatography (HPLC) conditions we referred to a published result [22]
120 rpm). 225 mL of the different kinds of designed media and cultured for 7 days (28 ± 1 °C, 120 containing rpm).In the static liquid culture stage, 25 mL mycelium suspension was transferred into a Blake bottle
Summary
Especially Ascomycetes, Deuteromycetes, and Basidiomycetes [1], have been attracting more and more attention in recent decades for their abundant bioactive secondary metabolites and their potential to be developed as drugs and health foods. It was reported that G. lucidum mainly contains two groups of bioactive components: polysaccharides and triterpenoids (GAs) [3]. The latter, when separated from G. lucidum, had anti-tumor activity both in vivo [4] and in vitro (95-D lung cancer cells, HCT-116 carcinoma cells, HeLa cells) [5,6,7,8], and anti-tuberculosis activity [8].The medicinal part of G. lucidum is its dried ripe fruiting body, which is highly lignified. Because of the great medicinal value of G. lucidum, liquid fermentation of the fungus was viewed as a promising way to produce polysaccharides and triterpenoids in a compact space and in a shorter time [11]. Structures of the five GA compounds isolated and used for content determination
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