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Aspergillus niger for largely producing xylanase and application thereof

A technology of xylanase and Aspergillus niger, applied in the direction of glycosylase, enzyme, hydrolase, etc., can solve the problems of low xylanase activity and rare enzymatic performance of xylanase, and achieve wide application Effect

Inactive Publication Date: 2014-07-16
GUANGZHOU INST OF ENERGY CONVERSION - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The study of de Souza Moreira (2013) showed that vanillin and ferulic acid have certain inhibitory or activating effects on xylanase from Aspergillus terreus, but the xylanase produced by Aspergillus terreus used in this study Very low enzyme activity
However, there is no report about the effects of furfural, acetic acid and 5-hydroxymethylfurfural on the xylanase enzymatic hydrolysis of lignocellulose after pretreatment.
In addition, ethanol produced during yeast fermentation can also inhibit the enzymatic hydrolysis of xylanase
[0005] There are very few reports on the effect of fermentation inhibitors on the hydrolysis performance of xylanase after pretreatment

Method used

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  • Aspergillus niger for largely producing xylanase and application thereof
  • Aspergillus niger for largely producing xylanase and application thereof
  • Aspergillus niger for largely producing xylanase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: the screening of bacterial strain

[0022] Screening samples are dead branches, rotten leaves, soil and other materials. After the screening samples are properly processed, they are serially diluted and applied to the xylan primary screening medium. The formula of the medium is: each liter of medium contains KH 2 PO 4 0.5g, (NH 4 ) 2 SO 4 2.0g, MgSO 4 ·7H 2 O0.25g, xylan (purchased from Shanghai Yuanju Biology) 5.0g, agar 18-20g, the balance is water, pH5.6. Culture at 30°C for 4-7d, pick the colony with larger hydrolysis transparent circle on PDA Purified on the culture medium, and the purified strains were stored on a PDA slant.

[0023] Re-screening of fermentation activity of xylanase-producing strains: inoculate the purified strains preserved on PDA slant into the same amount of fermentation re-screening culture medium, culture at 30°C, 120 rpm, for 6 days, and obtain fermentation liquid (ie, enzyme liquid). Based on the assay method of Bailey...

Embodiment 2

[0026] Embodiment 2: Preparation of xylanase and determination of its enzyme activity and enzymatic properties

[0027] 1. Preparation of xylanase preparation: Aspergillus niger (Aspergillus niger) SM751 was activated after seed cultivation. The seed medium was modified Mandel's nutrient solution (same as Example 1), added 5g / L xylan, and the initial pH was 5.6. Sterilize at 115°C for 30 minutes. The parameters of the shaker were set as follows: 30° C., 120 rpm, and the seed culture solution was obtained after culturing for 3 days. The seed culture solution was inoculated into the solid fermentation medium at an inoculum size of 10% (v / w). The formula of the solid-state fermentation medium is: the mass ratio of corncob to bran is 1:5, the mixture of corncob and bran is used as substrate and support material, and 1.4g / L (NH4) 2 SO 4 , 2.0g / L KH 2 PO 4 , 0.3g / L CaCl 2 , 0.3g / L MgSO 4 , adding an appropriate amount of trace elements (FeSO 4 ·7H 2 O, 5mg / L; CoCl 2 ,20mg...

Embodiment 3

[0033] Example 3: Effect of fermentation inhibitors on xylanase

[0034] 1. The inhibitory effect of a single fermentation inhibitor on xylanase (xylanase produced by Aspergillus niger (Aspergillus niger) SM751):

[0035] Different fermentation inhibitors have different inhibitory effects on xylanase. Such as Figure 4 It was shown that when the ethanol concentration was 10.00g / L, the enzyme activity residual rate of xylanase was 102.15%, and the enzyme activity increased by about 2.15%. When the concentration of acetic acid was 10.10g / L, the residual rate of xylanase activity was 99.05%. When the concentration of furfural was 1.40g / L, the residual rate of xylanase was 100.08%. When the concentration of 5-hydroxymethylfurfural was 1.20g / L, the residual rate of xylanase was 95.64%. When the concentration of vanillin was 1.10g / L, the residual rate of xylanase was 98.16%. When the concentration of ferulic acid was 1.10g / L, the residual rate of xylanase was 72.99%. Therefore...

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Abstract

The invention discloses aspergillus niger for largely producing xylanase and application thereof. The collection number of the Aspergillus niger SM751 is CGMCC No. 8670. The xylanase generated by the Aspergillus niger SM751 is activated in enzyme activity under the action of mixing of multiple inhibitors; compared with blank control, the maximal activation rate of the enzyme activity achieves 33.43%. The Aspergillus niger SM751 provided by the invention can be used for largely producing xylanase which has high enzyme activity and can tolerate the pretreated fermented inhibitors, thereby having wide application in the field of lignocellulose enzymolysis.

Description

Technical field: [0001] The invention belongs to the field of microbial fermentation and its enzyme engineering application, in particular to a high-yield xylanase Aspergillus niger (Aspergillus niger) SM751 and the produced xylanase, and the xylanase in lignocellulose hydrolysis Application of enzymatic hydrolysis of hemicellulose in liquid. Background technique: [0002] Global annual hemicellulose production is 6×10 10 Ton is an important renewable carbon resource bank second only to cellulose, which can be widely used to manufacture biofuels and many biochemical refining products. During the processing of plant raw materials rich in hemicellulose raw materials, adding xylanase can significantly accelerate the reaction process, promote the enzymatic hydrolysis effect of other enzymes such as cellulase, pectinase and mannanase, and significantly improve The usage characteristics of the product. Therefore, xylanase is widely used in traditional fields such as pulp and pa...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/42C12P19/14C12R1/685
CPCC12N9/248C12P19/14C12Y302/01008C12N1/145C12R2001/685
Inventor 袁振宏许敬亮何敏超张宇孔晓英陈小燕粱翠谊郭颖刘云云
Owner GUANGZHOU INST OF ENERGY CONVERSION - CHINESE ACAD OF SCI
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