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

Inactive Publication Date: 2011-08-17
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To sum up, whether it is liquid fermentation or solid-state fermentation, the activity of chitosanase produced by microorganisms is low at present, resulting in high production and application cos

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a method for producing chitosanase by using Aspergillus usamii, which belongs to the technical field of biological engineering. In the method, agricultural and sideline products such as bran, soybean cake powder and rapeseed cake powder are used as fermentation substrates, a proper amount of inorganic nitrogen source, chitosan, inorganic salt, surfactant and the like are added, and the chitosanase is produced by the solid fermentation of an Aspergillus usamii E001 strain. The invention provides the formula and culture conditions of a solid fermentation culture medium for test production of chitosanase by the Aspergillus usamii E001 in a laboratory, and the chitosanase activity of a mature fermented grain material of the medium reaches 1,942 to 2,461U/g. The method has the advantages of simple equipment, small investment, quick effectiveness, low production cost, environmentally-friendly, high activity of chitosanase fermentation enzyme, and the like, contributes to the development and utilization of chitosanase and has high economic and practical value.

Description

technical field [0001] The invention relates to a method for producing chitosanase by using Aspergillus usamii, in particular to a new process for producing chitosanase by solid-state fermentation of Aspergillus usamii E001 strain, which belongs to the technical field of bioengineering. Background technique [0002] Chitosan is the deacetylation product of chitin, and it is a polymer cationic flocculant. The chitosan oligosaccharide obtained by partial hydrolysis of chitosan has good water solubility, non-toxicity, no pyrogen, no variability, easy to be absorbed by biological organisms, and has unique physiological activity. Wide application prospects. Using chitosan, a biocatalyst with high specificity, to hydrolyze chitosan, it has the characteristics of mild reaction conditions, easy control of the reaction process, no pollution to the environment, the highest yield of chitosan oligosaccharides, and stable quality. It is an ideal production method. Chitooligosaccharide ...

Claims

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Application Information

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IPC IPC(8): C12N9/42
Inventor 邬敏辰史红玲吴静李剑芳陈伟唐存多
Owner JIANGNAN UNIV
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