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High xylanase yield Aspergillus niger and application thereof

A xylanase, Aspergillus niger technology, applied in the direction of enzymes, microorganism-based methods, and microorganism determination/inspection, etc., can solve problems such as low xylanase enzyme activity

Active Publication Date: 2014-08-20
杨凌未来中科环保科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As shown by de Souza Moreira (2013), vanillin and ferulic acid have a certain inhibitory effect on xylanase derived from Aspergillus terreus, but the xylanase activity of the Aspergillus terreus strain used is extremely low

Method used

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  • High xylanase yield Aspergillus niger and application thereof
  • High xylanase yield Aspergillus niger and application thereof
  • High xylanase yield Aspergillus niger and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: the screening of bacterial strain

[0023] 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.

[0024] 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 medium, culture at 30°C, 120 rpm, for 6 days. Based on the assay method of Bailey (1992) et al. with appropriate modifications. The specific...

Embodiment 2

[0028] Embodiment 2: Preparation of xylanase and determination of its enzyme activity and enzymatic properties

[0029] 1. Preparation of xylanase preparation: Aspergillus niger (Aspergillus niger) SM24 / a was activated, cultured in seeds, and the seed medium was modified Mandel's nutrient solution (same as in Example 1), and then xylan was added to make Its final concentration is 5g / L, pH5.6, and sterilized at 115°C for 30min. 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 liquid was inoculated into the solid fermentation medium with an inoculation amount of 7.5% (v / w). The formula of the solid fermentation medium was: the mass ratio of corncob to bran was 1:5, and the corncob and A mixture of substrate and support material, with 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 ...

Embodiment 3

[0035] Example 3: Tolerance of xylanase to fermentation inhibitors

[0036] 1. Tolerance of xylanase to a single inhibitor

[0037] The relative enzymatic activity of xylanase to different concentrations of the same fermentation inhibitor (such as ethanol, acetic acid, furfural, vanillin and ferulic acid) showed certain differences, but generally speaking, they all retained a higher Enzyme activity residual rate. When the concentrations of ethanol and acetic acid are 5g / L, 15g / L and 30g / L, and furfural (2.5g / L, 1.25g / L and 0.4g / L), ferulic acid (1.2g / L, 0.6g / L and 0.2g / L) and vanillin (1.3g / L, 0.65g / L and 0.2g / L) concentration range, the enzyme activity retention rate of xylanase of the present invention is 93~108%.

[0038] 2. Tolerance of xylanase to mixed inhibitors added with 5-hydroxymethylfurfural:

[0039] Such as Figure 4 As shown, in the mixed inhibitor system of higher concentration (the concentrations of ethanol, acetic acid, furfural, vanillin, ferulic acid a...

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Abstract

The present invention discloses high xylanase yield Aspergillus niger and an application thereof, wherein the Aspergillus niger SM24 / a has the preservation number of CGMCC No.8671. According to the present invention, the Aspergillus niger SM24 / a can produce xylanase providing good tolerance for ethanol, acetic acid, furfural, vanillin, ferulic acid and a mixture of the above materials, the xylanase production activity of the strain is high, the solid fermentation enzyme activity of the xylanase can be up to 10801 IU / g (catalysis production of 1 [mu]mol of xylose within 1 min is defined as an enzyme unit), the optimum pH value is 5.6, the optimum reaction temperature is 37 DEG C, and the 50 DEG C enzyme activity remaining rate is 79.01%; and 5-hydroxymethyl furfural provides an inhibition effect for the enzyme, and the concentration and the enzyme inhibition rate present a certain linear relationship when the 5-hydroxymethyl furfural exists in a separate or mixed manner, such that the xylanase can further be used as the qualitative even quantitative indicator of the 5-hydroxymethyl furfural concentration.

Description

Technical field: [0001] The invention belongs to the field of microbial fermentation and its enzyme engineering application, and in particular relates to a high-yield xylanase Aspergillus niger SM24 / a and the rapid detection of 5-hydroxymethyl by the enzyme inhibition method used in the preparation of the xylanase application of formulations based on furfural concentrations. Background technique: [0002] Xylanase, the English name xylanase, referred to as XYL, is an enzyme that can catalyze the hydrolysis of xylopyranose in the glycosidic bond of β-1,4 xylan, thereby releasing xylose and xylooligosaccharides. In the processing of hemicellulose raw materials and even cellulose raw materials, adding xylanase can significantly accelerate the reaction process, improve the use characteristics of products, and reduce the cost, energy consumption and environmental pollution of the production process. Therefore, xylanase is widely used in traditional fields such as pulp and paper,...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/42C12Q1/34C12R1/685
CPCC12N9/248C12Q1/34C12N1/145C12R2001/685G01N2333/924
Inventor 袁振宏许敬亮何敏超张宇孔晓英粱翠谊陈小燕刘云云刘颖
Owner 杨凌未来中科环保科技有限公司
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