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Process of producing beta-mannase with Aspergillus niger

A technology of mannanase and solid-state fermentation, applied in the field of enzyme engineering, can solve the problems of high price and hindering the wide-scale application of β-mannanase

Inactive Publication Date: 2009-11-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the high price hinders the wide application of β-mannanase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Strain: Aspergillus niger WA9024.

[0028] Erlenmeyer flask solid-state fermentation medium: 10g base material in 250mL Erlenmeyer flask (including: bran 6.4g, corn flour 1.6g, rapeseed cake flour 2.0g), konjac flour 0.4g, corn steep liquor 0.5g, (NH 4 ) 2 SO 4 100mg, KH 2 PO 4 30mg, CaCl 2 10mg, CuSO 4 2H 2 O 0.5mg, KI 0.5mg, MnSO 4 ·H 2 O 0.5mg, tap water 17mL, pH 6.0.

[0029] Conditions for solid-state fermentation in a triangular flask: sterilize the medium at 121°C for 40 minutes, inoculate one fungus WA9024 slant seed after cooling, and culture it at 32°C for 84 hours. 1.7 mL sterile water.

[0030] The test results of the above-mentioned Erlenmeyer flasks repeated for 5 batches showed that the average enzyme activity of β-mannanase in the mature fermented koji material reached 22185IU / g dry koji.

Embodiment 2

[0032] Strain: Aspergillus niger WA9024.

[0033] Industrial production of solid-state fermentation equipment: 30M 3 Solid-state fermenter with automatic temperature control blowing device, mechanical turning device, mechanical water replenishing device and mechanical discharging device, etc. The diameter of the tank is 4.5m, the height of the tank is 2.0m, and 1000kg of base material (calculated as dry material) is actually loaded, and the accumulation thickness of the fermentation medium is 30cm.

[0034] Industrialized production of solid-state fermentation medium: 1000kg base material (among them: bran 800kg, corn flour 100kg, rapeseed cake flour 100kg), konjac flour 40kg, corn steep liquor 50kg, (NH 4 ) 2 SO 4 10kg, KH 2 PO 4 3kg, CaCl 2 1.0kg, CuSO 4 2H 2 O 0.05kg, KI 0.05kg, MnSO 4 ·H 2 O 0.05kg, tap water 2000kg, pH 6.0.

[0035] Industrial production solid-state fermentation conditions: add the above fermentation medium to 6M 3 In a high-pressure cooker...

Embodiment 3

[0038] In order to facilitate the growth of bacteria, achieve the purpose of shortening the fermentation cycle and controlling the contamination of bacteria, this embodiment adopts a variable temperature control mode in which the temperature of the solid-state fermentation is high before and then low, and the formula of the solid-state fermentation medium is the same as that of Example 2.

[0039] After the solid-state fermentation medium is sterilized, cooled and inoculated, it is placed at 30M 3 solid state fermenter. (1) In the early stage of fermentation (0-20h): 0-8h, close the air inlet and outlet, and set the temperature controller at 36°C; 8-12h, open 1 / 3 of the air inlet and outlet, and then gradually Open the air inlet and outlet until they are all open, and the temperature is still controlled at 36°C. (2) Middle stage of fermentation (20-56h): Turn the koji once every 20 hours and add 200kg of sterile water at the same time, and lower the set temperature to 35°C; t...

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PUM

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Abstract

The invention discloses a method for producing β-mannanase by using Aspergillus niger, which belongs to the technical field of enzyme engineering. The present invention adopts agricultural and sideline products such as bran, corn flour and rapeseed cake powder as base material, and adds appropriate amount of konjac flour, corn steep liquor, inorganic salt and tap water etc., utilizes Aspergillus niger (Aspergillus niger) WA9024 bacterial strain solid-state fermentation to produce Mannanase provides the fermentation medium formula and fermentation conditions for small-scale production and industrial production. The β-mannanase activity of mature fermented koji produced in small-scale production reaches 18912-22185IU / g dry koji, which is higher than that of the original koji WA9024 strain. The enzyme activity (8984IU / g dry koji) of small-scale production of solid-state fermentation has increased by 2.11-2.47 times, which is higher than the current domestic level; the enzyme activity of industrial production products is 12678-15921IU / g dry koji. The present invention uses cheap bran, corn flour, rapeseed meal and the like as main raw materials for fermentation, is a kind of fully utilizing natural renewable resources, and is also a green and sustainable production method.

Description

technical field [0001] The invention relates to a method for producing beta-mannanase by solid-state fermentation of Aspergillus niger WA9024 strain, which uses agricultural and sideline products such as bran, corn flour and rapeseed cake powder as main raw materials, and belongs to the technical field of enzyme engineering. technical background [0002] β-1,4-D-mannan mannohydrolase (β-1,4-D-mannan mannohydrolase, EC3.2.1.78) is a kind of enzyme derived from mannan, glucomannan, galactomannan The hydrolase that randomly cuts the β-1,4-D-mannosidic bond inside the main chain of sugar and galactoglucomannan, etc., referred to as β-mannanase (β-mannanase), belongs to the hemicellulase class. Widely present in lower animals, legumes, marine organisms and microorganisms. [0003] With the development and utilization of natural hemicellulase, the in-depth understanding of the anti-nutritional factors of β-mannan and the continuous discovery of the physiological functions of mann...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12N1/14C12R1/685
Inventor 邬敏辰李剑芳张树飞李江华徐春梅盛金萍邓珊珊方诗月
Owner JIANGNAN UNIV
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