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Biomass and extracellular polysaccharide of Ganoderma tsugae have various biological activity including anti-inflamatory activity, antioxidant activity and antitumor activity. However, the growth rate of G. tsugae in nature is very slow. Therefore, many studies have attempted to develop mass culture systems for G. tsugae using laboratory techniques. Many parameters of submerged fermentation for G. tsugae were studies to determine the optimization of process by combination of statistical techniques. Ten parameters from preliminary results and literature reviews (maltose, skim milk, KH2PO4+K2HPO4, MgSO4-7H2O, CaCO3, vitamin B5+B6, olive oil, ethanol, pH and shaking speed) were screened by Packett Berman design. The significant parameters were determined the optimal ranges by path of steepest ascent method. The optimal condition of process was performed by response surface method. Maltose, skim milk and pH are significant parameters for G. tsugae cultivation. The conditions of 31.031 g L-1 maltose, 14.055 g L-1 skim milk and an initial pH of 7.12 resulted in the maximum extracellular polysaccharide content of 415 mg L-1 and the same fermentation broth at an initial pH of 6.46 exhibited the most biomass at 15.776 g L-1. Finally, the optimal condition was compared with un-optimal condition which result indicates that the combination of statistical techniques enhance the productions of biomass and extracellular polysaccharide (13X and 1.5X of the control, respectively). Therefore, these strategies are useful for improvement of submerged fermentation of G. tsugae which it can apply in pharmaceutical industry.

The quantitative effects of oxygen supply in terms of shaking speed and medium volume on the production of mycelia and extracellular polysaccharide (EPS) using Ganoderma tsugae in submerged fermentation were investigated. Mycelia growth required the proper shaking speed at 134 rpm for breaking a larger pellet into several smaller pellets. Furthermore, high level of medium volume for consumption of abundant nutrient is needed for maximum mycelia growth. For EPS, a high agitation is needed to promote a good mass transfer for achieving high product concentrations recovery. The appropriate medium volume was found to be 150.4 mL for aiding to produce a secondary metabolite by promoting the mass transfer of substrates. The dry cell mass (DCM)-EPS diagram of G. tsugae was created from the both production equations which this diagram is an useful tool for submerge fermentation industry for decision about to control the both productions.