Gan-Rong Xu

Enhanced production of pigments by addition of surfactants in submerged fermentation of Monascus purpureus H1102.

BACKGROUND The production of pigments by Monascus spp. has attracted increasing attention. Modification of the cell membrane structure by addition of surfactants has proved to be effective for the secretion of intracellular metabolites. Hence in this study the effects and underlying mechanism of surfactants on the production of pigments in submerged fermentation of Monascus purpureus H1102 were systematically investigated. RESULTS Various surfactants exerted significant but different impacts on the biomass and production of pigments. The maximum production of pigment (304.3 U mL(-1) ) and highest extracellular/intracellular pigment ratio (1.46) were achieved when 15 g L(-1) Triton X-100 was added at 24 h of fermentation, corresponding to significant increases of 88.4 and 240% respectively compared with the control. Meanwhile, the concentration of citrinin (0.94 mg L(-1) ) was 20.6% lower than that of the control. A further study on the fatty acid composition of M. purpureus H1102 showed that the unsaturated/saturated fatty acid ratio and the index of unsaturated fatty acid increased significantly with the addition of Triton X-100. CONCLUSION The addition of surfactant Triton X-100 could greatly enhance the production of pigment. It was suggested that Triton X-100 facilitated the secretion of intracellular pigment and therefore enhanced pigment production accordingly.

Why solid-state fermentation is more advantageous over submerged fermentation for converting high concentration of glycerol into Monacolin K by Monascus purpureus 9901: A mechanistic study.

The underlying mechanisms by which solid-state fermentation (SSF) was more advantageous over submerged fermentation (SmF) for converting high concentration of glycerol into Monacolin K by Monascus purpureus were investigated innovatively. First, the established kinetic models and kinetic parameters showed that the cell growth, Monacolin K formation and glycerol consumption in SSF were more rapid than those in SmF. Secondly, the comparison of fatty acid composition of mycelial cells indicated a better fluidity and permeability of the cell membrane in SSF than that of SmF, which was also consistent with the difference in the ratio of extracellular/intracellular Monacolin K between the two systems. Thirdly, the phenomenon of glycerol concentration gradient was verified in SSF, which could well explain the resistance effect to high concentration of glycerol in SSF. These new findings provide some important insights to the elucidation of the advantages of SSF for the synthesis of fungal secondary metabolites.

Changes in volatile compound composition of Antrodia camphorata during solid state fermentation.

BACKGROUND Although the volatiles present in mushrooms and fungi have been investigated by many researchers, including Antrodia camphorata in submerged fermentation, there are few data available regarding changes in volatile compounds during fermentation. Our research has revealed that solid state fermentation of A. camphorata is highly odiferous compared with submerged cultures and the odor changed with increasing culture time. Therefore the aim of this study was to investigate the changes in volatile compound composition of A. camphorata during solid state fermentation. RESULTS Altogether, 124 major volatile compounds were identified. The volatile compounds produced by A. camphorata during growth in solid state fermentation were quite different. Oct-1-en-3-ol, octan-3-one and methyl 2-phenylacetate were predominant in exponential growth phase production, while the dominant volatiles produced in stationary phase were octan-3-one and methyl 2-phenylacetate. In stationary phase, lactone compounds in A. camphorata, such as 5-butyloxolan-2-one, 5-heptyloxolan-2-one, 6-heptyloxan-2-one, contributed greatly to peach and fruit-like flavor. Terpene and terpene alcohol compounds, such as 1-terpineol, L-linalool, T-cadinol, (E, E)-farnesol, β-elemene, cis-α-bisabolene and α-muurolene, made different contributions to herbal fresh aroma in A. camphorata. Nineteen volatile sesquiterpenes were detected from solid state fermentation of A. camphorata. The compounds 5-n-butyl-5H-furan-2-one, β-ionone, (-)-caryophyllene oxide, aromadendrene oxide, diepi-α-cedrene epoxide, β-elemene, α-selinene, α-muurolene, azulene, germacrene D, γ-cadinene and 2-methylpyrazine have not hitherto been reported in A. camphorata. CONCLUSION The preliminary results suggest that the aroma-active compounds produced by A camphorata in solid state fermentation might serve as an important source of natural aroma compounds for the food and cosmetic industries or antibiotic activity compounds. The sesquiterpenes could be identified as possible taxonomic markers for A. camphorata.

Stimulating the biosynthesis of Antroquinonol by addition of effectors and soybean oil in submerged fermentation of Antrodia camphorata.

Antrodia camphorata is a precious medicinal mushroom that has attracted increasing attentions. Antroquinonol has been considered as being among the most biologically active components of A. camphorata. However, it was hardly biosynthesized via conventional submerged fermentation. Two approaches were applied to stimulate the biosynthesis of antroquinonol in submerged fermentation. On one hand, different kinds of effectors that may involve in the antroquinonol biosynthesis were investigated. Among the tested effectors, camphorwood leach liquor was the most effective for stimulating the antroquinonol production. On the other hand, because of the hydrophobic characteristics of antroquinonol, soybean oil was added to establish an extractive fermentation system for alleviating the product inhibition and resulting in enhanced productivity. The highest antroquinonol concentration could be achieved at 89.06 ± 0.14 mg/L when 10% (v/v) soybean oil was added at the beginning of the fermentation. This study will be of great significance for the study of A. camphorata and the bioprocess regulation of antroquinonol production.

Enhanced production of Monacolin K by addition of precursors and surfactants in submerged fermentation of Monascus purpureus 9901

The main problem in Monacolin K (MK) production by submerged fermentation of Monascus purpureus is low productivity. In this study, on one hand, addition of precursors was used to activate the biosynthesis of MK. When 4.0 g/L of sodium citrate was supplemented at the 48th H of the fermentation, the final MK production reached to 1,658.9 ± 28.5 mg/L after 20 day of fermentation, which was improved by 52.6% compared with that of the control. On the other hand, addition of surfactants could increase the permeability of cell membrane, thus driving more intracellular metabolites secreted into the fermentation broth and alleviating the product inhibition. When 40.0 g/L of Triton X‐100 was added at the beginning of the fermentation, the final MK production reached to 2,026.0 ± 30.4 mg/L after 20 day of fermentation, which was improved by 84.9% compared with that of the control. These results are helpful to provide some new insights into the biosynthetic regulation on MK production; the approach can be applied to other fungal fermentation processes for enhancing production of useful metabolites.

Induction of Antroquinonol Production by Addition of Hydrogen Peroxide in the Fermentation of Antrodia camphorata S‐29

BACKGROUND Antroquinonol have significantly anti-tumour effects on various cancer cells. There is still lack of reports on regulation of environmental factors on antroquinonol production by Antrodia camphorata. RESULTS An effective submerged fermentation method was employed to induce antroquinonol with adding H2 O2 . The production of antroquinonol was 57.81 mg L-1 after fermentation for 10 days when adding 25 mmol L-1 H2 O2 at day 4 of the fermentation process. Then, antroquinonol was further increased to 80.10 mg L-1 with cell productivity of 14.94 mg g-1 dry mycelium when the feeding rate of H2 O2 was adjusted to 0.2 mmol L-1 h-1 in the 7 L fermentation bioreactor. After inhibiting the generation of reactive oxygen species with the inhibitor diphenyleneiodoium, the synthesis of antroquinonol from A. camphorata was significantly reduced, and the yield was only 3.3 mg L-1 . CONCLUSION The results demonstrated that addition of H2 O2 was a very effective strategy to induce and regulate the synthesis of antroquinonol in submerged fermentation. Reactive oxygen species generated by H2 O2 during fermentation caused oxidative stress, which induced the synthesis of antroquinonol and other chemical compounds. Moreover, it is very beneficial process to improve production and diversity of the active compounds during liquid fermentation of A. camphorata mycelium.

Current Advances on the Structure, Bioactivity, Synthesis, and Metabolic Regulation of Novel Ubiquinone Derivatives in the Edible and Medicinal Mushroom Antrodia cinnamomea

In recent years, Antrodia cinnamomea has attracted great attention around the world as an extremely precious edible and medicinal mushroom. Ubiquinone derivatives, which are characteristic metabolites of A. cinnamomea, have shown great bioactivities. Some of them have been regarded as promising therapeutic agents and approved into clinical trial by the U.S. Food and Drug Administration. Although some excellent reviews have been published covering different aspects of A. cinnamomea, this review brings, for the first time, complete information about the structure, bioactivity, chemical synthesis, biosynthesis, and metabolic regulation of ubiquinone derivatives in A. cinnamomea. It not only advances our knowledge on the bioactive metabolites, especially the ubiquinone derivatives, in A. cinnamomea but also provides valuable information for the investigation on other edible and medicinal mushrooms.

Efficient Biosynthesis of Natural Yellow Pigments by Monascus purpureus in a Novel Integrated Fermentation System

Because of the increasing demand for healthy and safe food, Monascus spp. have gained much attention as a sustainable source of natural food colorant. In this study, a novel integrated fermentation system consisting of surfactant and in situ extractant was established for efficiently producing yellow pigments by M. purpureus sjs-6. The maximum production of Monascus yellow pigment (669.2 U/mL) was obtained when 40% soybean oil (as extractant) was supplied at the beginning and 5 g/L Span-80 (as surfactant) was supplied at the 72nd h, which resulted in production 27.8-times of that of the control. Critical factors such as alleviating the product inhibition, increasing the membrane permeability, changing the hyphal morphology, and influencing the cell activity have been suggested as the underlying mechanisms. This system is of great significance for the bioprocess, which suffers product inhibition, and it can serve as a promising step for enhancing the yield of hydrophobic metabolites.

Effect of cultural conditions on antrodin C production by basidiomycete Antrodia camphorata in solid‐state fermentation

Antrodia camphorata is a medicinal fungus and antrodin C is one of the main bioactive components of A. camphorata in the submerged fermentation (SmF). To optimize the culture conditions, the factors influencing the production of antrodin C by A. camphorata under solid‐state fermentation (SSF) were investigated in this study. Different solid substrates and external nitrogen sources were tested for their efficiency in producing antrodin C. The response surface methodology was applied to evaluate the influence of several variables, namely, the concentrations of soybean meal, initial moisture content, and inoculum density on antrodin C production in solid‐state fermentation. The experimental results show that the optimum fermentation medium for antrodin C production by A. camphorata was composed of 0.578 g soybean meal, 0.05 g Na2HPO4, 0.05 g MgSO4 for 100 g rice, with 51.83% initial moisture content, 22 day culture time, 28 °C culture temperature, and 35.54% inoculum density. At optimized conditions, 6,617.36 ± 92.71 mg kg−1 yield of antrodin C was achieved. Solid‐state fermentation is one good cultural method to improve the production of antrodin C by A. camphorata.