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Endoxylanase and application thereof in xylose production

A sequence table, protein technology, application in the production of xylobiose, endoxylanase and its application in the production of xylobiose, protein field, can solve the high requirements of equipment and high energy consumption , unfriendly environment and other issues

Active Publication Date: 2019-01-15
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the chemical hydrolysis method requires the use of high-concentration chemicals (such as strong acid and strong alkali), it has the disadvantages of being unfriendly to the environment and requiring high equipment and equipment; the self-hydrolysis method requires the use of high temperature (generally much higher than 100ºC) and high pressure, so there is a problem of energy consumption. However, the enzymatic hydrolysis method is carried out under mild conditions, so it is an environmentally friendly and low-energy method (Mano MCR, Neri-Numa IA, da Silva JB, Paulino BN, Pessoa MG, et al. al. (2018) Oligosaccharide biotechnology: an approach of prebiotic revolution on the industry. Appl Microbiol Biotechnol 102: 17–37.)

Method used

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  • Endoxylanase and application thereof in xylose production
  • Endoxylanase and application thereof in xylose production
  • Endoxylanase and application thereof in xylose production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1. Gene SipoEnXyn10A Gain

[0063] (1) Extraction of genomic DNA of Streptomyces sweetpotato

[0064] The Streptomyces sphaeroides was inoculated to the modified Gao's No. 1 solid medium plate, and incubated at 28°C for 5 days to obtain fresh and vigorous spores. The spores were inoculated into Gao's No. 1 liquid medium (50 mL in a 250 mL Erlenmeyer flask), and cultured on a shaker at 28°C and 180 rpm for 2 days. The grown mycelium can be used to extract genomic DNA.

[0065] Use the genomic DNA rapid extraction kit to extract the genomic DNA of Streptomyces sweetpotato. The operation is carried out in accordance with the product instructions. The extracted DNA solution was subjected to 0.8% agarose gel electrophoresis, and the results were as follows figure 1 Shown. figure 1 The sample in the middle lane 1 is a 1 kb ladder, containing 7 DNA fragments, from large to small, respectively 15, 10, 7.5, 5, 2.5, 1 and 0.25 kb; the sample in lane 2 is the genomic DNA of S...

Embodiment 2

[0071] Example 2. Functional verification of the recombinant protein RSipoEnXyn10A

[0072] 1. Recombinant expression vector pET30a(+)- RSipoEnXyn10A Build

[0073] Use restriction enzymes for vector pET30a (+) Eco RⅠand Xho Ⅰ Double enzyme digestion, the product of the double enzyme digestion was recovered with the DNA purification and recovery kit and then tested by 0.8% agarose gel electrophoresis. There was only one 5.4 kb long vector band in the lane ( image 3 A, The sample in lane 1 is a 1 kb DNA ladder, containing 14 DNA fragments, from large to small, 10, 8, 6, 5, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.75, 0.5 and 0.25 kb; the sample in lane 2 uses restriction enzymes Eco R Ⅰ and Xho Ⅰ The empty vector pET30a (+) is double-enzyme-cut, and then the product is recovered by DNA purification and recovery kit).

[0074] The PCR product with sequence 1 obtained in Example 1 was subjected to 0.8% agarose gel electrophoresis, the gel block where the band was located was cut off, and the ...

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PUM

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Abstract

The invention relates to the biochemical field, in particular to a protein RSipoEnXyn10A and its application as an endoxylanase in the production of xylose. The protein RSipoEnXyn10A is a protein represented by the following 1) or 2): 1) a protein composed of an amino acid sequence represented by SEQ ID NO: 3 in the sequence list; 2) a protein derive from 1) which is formed by substitute, deletingor adding one or more amino acid residues to that amino acid sequence shown in SEQ ID NO: 3 in the sequence table and has the same function. The invention obtains a gene SipoEnXyn10A encoding an endoxylanase from the genome sequence of Streptomyces suavissimus, the gene can be expressed in host cells to produce the protein RSipoEnXyn10A. The protein RSipoEnXyn10A is used as an endoxylanase in theproduction of xylenedisaccharide. The endoxylanase is a kind of endoxylanase with the properties of high temperature, neutrality and xylenedisaccharide release from xylan hydrolyzed from agroforestryresidues, which is of great significance to the production of xylenedisaccharide.

Description

Technical field [0001] The present invention relates to the field of biochemical industry, in particular to an endoxylanase and its application in the production of xylobiose, in particular to a protein RSipoEnXyn10A and its use as an endoxylanase in agricultural and forestry residues The application of raw materials in the production of xylobiose. Background technique [0002] Xylooligosaccharides refer to the linear oligomers of xylose linked by β-(1,4) xylosidic bonds. Generally, molecules with a degree of polymerization of 2 to 20 xylose are considered to be xylo-oligosaccharides. Molecules greater than 20 are xylan (Carvalho AFA, deOliva Neto P, da Silva DF, Pastore GM (2013) Xylo-oligosaccharides from lignocellulosic materials: Chemical structure, health benefits and production by chemical and enzymatic hydrolysis. Food Res Int 51: 75– 85.). The human body cannot digest xylo-oligosaccharides, so xylo-oligosaccharides are called non-digestible oligosaccharides (NDOs). Xyl...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N15/56C12N15/70C12N1/21C12P19/12C12R1/19
CPCC12N9/2482C12N15/70C12P19/12C12Y302/01008
Inventor 刘幽燕冼亮唐爱星覃益民李青云黎忠
Owner GUANGXI UNIV
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