Method for producing chitosanase by using Aspergillus usamii
A technology of Aspergillus Usami and chitosanase, which is applied in the field of bioengineering, can solve the problems of high production and application costs of chitosanase, low activity of chitosanase, and restrictions on wide application, etc., and achieve low production cost and simple equipment , Environmentally friendly effect
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Embodiment 1
[0017] Strain: Aspergillus usamii E001 strain.
[0018] Solid-state fermentation medium: 10g of base material (9g of bran + 1g of bean cake powder) in a 250mL triangular bottle, (NH 4 ) 2 SO 4 100mg, MgSO 4 10mg, CaCl 2 10mg, KH 2 PO 4 50mg, tap water 12mL, pH 6.0.
[0019] Solid-state fermentation conditions: sterilize the medium at 121°C for 40 minutes, inoculate one fungus E001 bran koji seeds after cooling, and culture at 30°C for 84 hours, during which time the koji is turned once at 20 hours and 40 hours respectively.
[0020] Repeat the above-mentioned Erlenmeyer Flask experiment result of 3 batches (do 3 bottles in parallel in each batch) and show that the chitosanase production average enzyme activity of E001 mature solid-state fermented koji material reaches 1942U / g dry koji.
Embodiment 2
[0022] Strain: Aspergillus usamii E001 strain.
[0023] Solid-state fermentation medium: 10g base material (8g bran, 1g bean cake powder, 1g rapeseed cake powder) in 250mL triangular bottle, 0.2g chitosan, (NH 4 ) 2 SO 4 100mg, Tween-80 40mg, MgSO 4 10mg, CaCl 2 10mg, KH 2 PO 4 50mg, tap water 14mL, pH 6.0.
[0024] Solid-state fermentation conditions: sterilize the medium at 121°C for 40 minutes, inoculate one fungus E001 bran koji seeds after cooling, and culture at 30°C for 96 hours, during which time the koji is turned once at 20 hours and 40 hours respectively.
[0025] Repeat the above-mentioned Erlenmeyer flask experiment result of 3 batches (do 3 bottles in parallel for each batch) and show that the chitosanase average enzyme activity of E001 mature solid-state fermented koji material reaches 2274U / g dry koji.
Embodiment 3
[0027] In order to facilitate the growth of the bacteria, shorten the fermentation period and control the contamination of bacteria, this embodiment adopts the variable temperature control mode of the solid state fermentation temperature, which is high at the beginning and then low.
[0028] After the solid-state fermentation medium is sterilized, cooled and inoculated, it is cultivated at 34°C for 16 hours, and the song is turned for the first time; then it is cultivated at 32°C for 36 hours, and the song is turned for the second time; finally, it is cultivated at 30°C for 72 hours.
[0029] The above-mentioned Erlenmeyer flask experiment results of repeating 3 batches (3 bottles in parallel for each batch) showed that the chitosanase average enzyme activity of E001 mature solid-state fermented koji material reached 2461U / g dry koji.
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