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Most factories use liquid spawns for efficient, industrialized, and standardized cultivation, addressing intensive production bottlenecks.  

1. Types and Selection of Shakers

 - Reciprocating shakers: 80-140 r/min frequency, 5-14 cm stroke; lower cost, suitable for small factories; risk of liquid splashing and contamination if parameters are improper.   - Rotary shakers: 3-6 cm eccentricity, 60-300 r/min speed; complex structure, higher cost, better oxygen transfer, lower power consumption, no liquid splashing; used in large factories/laboratories.

 2. Shake Flask Culture Process Flow

 Prepare medium → Package → Sterilize → Cool → Inoculate → Shake flask culture.  

3. Key Points for Shake Flask Production

 - Medium formula: 200g potatoes, 20g glucose, 5g peptone, 2.0g KH₂PO₄, 1.0g MgSO₄, vitamin B1, pH 6.8-7.0, 1000ml water.   - Medium preparation: Boil potatoes, filter, add ingredients (peptone dissolved in cold water first), adjust pH (slightly higher before sterilization as it drops after autoclaving).   - Packaging: Pour into Erlenmeyer flasks (50ml in 250-300ml flasks; 150ml in 500ml flasks) with 10 glass beads; seal with silicone breathable stoppers (or 12-layer gauze + kraft paper/newspaper).   - Sterilization and cooling: Autoclave at 121°C for 30 minutes; cool to below 30°C, place in clean bench.   - Inoculation: Use fresh slant cultures; inoculate 2-3 cm² mycelia per flask under sterile conditions; 4-5 flasks per slant; label.   - Cultivation: Static incubation at 25°C for 48h, then shake culture (100 rpm for reciprocating, 150 rpm for rotary shakers) at 25°C for 5-7 days. Qualified when: no contamination, hyphae dry weight 1.5g/100ml, mycelial pellets 1-2mm in diameter; use immediately to avoid aging.   - Storage: Store at 4°C.   - Quality requirements: Clear medium, dense brown mycelial pellets (like millet porridge), no impurities/autolysis/wall detachment, sweet smell; no foreign bacteria under microscopy.