The complex scheme of unit operations shown in Fig.1. for simultaneous extraction and purification of enzymes, lectins, free and protein- bound polysaccharides with triple helix structure of β-1,3-1,6-glucan and secondary metabolites including terpenes, alkaloids and antibiotics from either spent mushroom compost, mushroom stems or partially degraded mushroom fruiting bodies as follows: Pyranose dehydrogenase, xylanase, pectinase, protease and laccase from spent mushroom compost of Agaricus bisporus defined as retentate in Fig. 1 are separated by immobilized metal affinity chromatography (IMAC) by using epoxy- activated Sepharose 6B- IDA- Cu as chromatographic matrix and 20 mM phosphate buffer pH 6.0 or 7.0 containing 1M NaCl as mobile phase. Tyrosinase, superoxide dismutase and protein-bound polysaccharides with triple helix structure of β-1,3-1,6--glucan from partially degraded mushroom fruiting bodies defined as retentate in Fig.1 are separated by immobilized metal affinity chromatography using epoxy- activated Sepharose 6B- IDA- Cu as chromatographic matrix and 20 mM phosphate buffer pH 6.0 or 7.0 containing 1 M NaCl, as mobile phase. Extracellular free and protein-bound polysaccharides with triple helix structure of β-1,3-1,6--glucan from spent mushroom compost defined as Sediment III in Fig.1 are separated by extraction, precipitation and gel-filtration chromatography by using Sephacryl S-300 HR-300 or 100 and 20 mM phosphate buffer pH 6.0 or 7.0, respectively, as chromatographic matrix and mobile phase, respectively. Terpenes, alkaloids and antibiotics with biological activities were separated by extraction and HPLC from spent mushroom compost defined as aqueous phase I in Fig. 1 by Nucleosil 100-7 C-18 250 x 4.6 mm i.d. with a guard column of CC 8/4 Nucleosil 100-5 C18 and methanol and water (0.5:1), as chromatographic matrix and mobile phase, respectively. Pyranose dehydrogenase, xylanase, pectinase, protease and laccase from spent mushroom compost of Agaricus bisporus defined as retentate in Fig. 1 are separated by immobilized metal affinity chromatography (IMAC) by using epoxy- activated Sepharose 6B- IDA- Cu as chromatographic matrix and 20 mM phosphate buffer pH 6.0 or 7.0 containing 1M NaCl as mobile phase. Tyrosinase, superoxide dismutase and protein-bound polysaccharides with triple helix structure of β-1,3-1,6--glucan from partially degraded mushroom fruiting bodies defined as retentate in Fig.1 are separated by immobilized metal affinity chromatography using epoxy- activated Sepharose 6B- IDA- Cu as chromatographic matrix and 20 mM phosphate buffer pH 6.0 or 7.0 containing 1 M NaCl, as mobile phase. Extracellular free and protein-bound polysaccharides with triple helix structure of β-1,3-1,6--glucan from spent mushroom compost defined as Sediment III in Fig.1 are separated by extraction, precipitation and gel-filtration chromatography by using Sephacryl S-300 HR-300 or 100 and 20 mM phosphate buffer pH 6.0 or 7.0, respectively, as chromatographic matrix and mobile phase, respectively. Terpenes, alkaloids and antibiotics with biological activities were separated by extraction and HPLC from spent mushroom compost defined as aqueous phase I in Fig. 1 by Nucleosil 100-7 C-18 250 x 4.6 mm i.d. with a guard column of CC 8/4 Nucleosil 100-5 C18 and methanol and water (0.5:1), as chromatographic matrix and mobile phase, respectively.