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Strain for directly producing butanol by utilizing xylan as sole carbon source and application of strain

A technology for producing butanol and xylan, applied in the field of microorganisms, can solve problems such as increased production cost, unfavorable industrial production and the like

Active Publication Date: 2017-08-01
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because it cannot use lignocellulose as a carbon source, its production cost is greatly increased, which is not conducive to industrial production

Method used

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  • Strain for directly producing butanol by utilizing xylan as sole carbon source and application of strain
  • Strain for directly producing butanol by utilizing xylan as sole carbon source and application of strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation and screening of thermophilic strain Thermoanaerobacterium thermosaccharolyticum M5 using xylan as the sole carbon source:

[0029] Weigh 1 g of soil samples taken from Inner Mongolian haystacks, dilute them with physiological saline, draw 200 μL onto a plate with xylan as the sole carbon source, and culture them anaerobically at 60°C for 5 days. The grown colonies were streaked and purified for 5 generations to screen out the strains that can directly utilize xylan and are thermophilic.

[0030] The medium formula of above-mentioned flat plate is NaCl 1g / L, K2 HPO 4 0.75g / L, KH 2 PO 4 0.75g / L, yeast powder 3g / L, CaCl 2 ·2H 2 O 0.015g / L, FeCl 2 ·4H 2 O 1.5g / L, KCl 0.3g / L, adjust pH to 6.5, xylan 30g / L, add agar powder 15-20g / L to solid medium, pass nitrogen for 10-20min, sterilize at 121°C for 15min.

Embodiment 2

[0032] Identification and growth characteristics of a thermophilic strain Thermoanaerobacterium thermosaccharolyticum M5 using xylan as the sole carbon source:

[0033] Identification of M5:

[0034] 16S rDNA identification of DL-10: The 16S rDNA of strain M5 was amplified using primers 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-TACCTTGTTACGACTT-3', and ligated to the cloning vector pMD19T by means of T / A cloning , construct the recombinant cloning vector pMD19T-16S, transform it into the cloned host strain Escherich coli DH5α to obtain the recombinant microorganism Escherich coli DH5α (pMD19T-16S), sequence the obtained exogenous fragments of the recombinant microorganism, and compare the 16S rDNA in the NCBI database. Sequence, the strain M5 was identified to the genus Thermoanaerobacterium thermosaccharolyticum at the molecular level, and the nucleotide sequence of its 16S rDNA is shown in SEQ ID NO: 1 in the sequence listing.

[0035] M5 Growth and Metabolic Properties...

Embodiment 3

[0038] The thermophilic strain Thermoanaerobacterium thermosaccharolyticum M5 using xylan as the sole carbon source utilizes different carbon sources for growth and fermentation characteristics:

[0039] The strain Thermoanaerobacterium thermosaccharolyticum M5 can use glucose, xylose, starch, xylan and other monosaccharides or polysaccharides as carbon sources for growth ( figure 1 ). Strain Thermoanaerobacterium thermosaccharolyticum M5 Pick a single colony of strain M5 from the plate and inoculate it into 5ml fermentation medium, 60℃, 120r·min -1 Cultured for 48h, then inoculated into the fermentation medium with an inoculum volume of 5% v / v, 60°C, 120r·min -1 The culture was shaken, the pH was adjusted to 6.0-6.2 every 24h, and the concentrations of various products were measured by GC after 72h. The concentration of fermentation products after 120h fermentation was as follows: ethanol 2.52g / L, butanol 0.78g / L, acetic acid 3.34g / L, butyric acid 3.05g / L.

[0040] Above-m...

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Abstract

The invention discloses a strain for directly producing butanol by utilizing xylan as a sole carbon source. A classification name of the strain is Thermoanaerobacterium thermosaccharolyticum, and the number of the strain is M5; moreover, the strain is preserved in China Center for Type Culture Collection (CCTCC), the preservation date of the strain is 27th February, 2017, and the preservation number is CCTCC NO: M 2017072. The strain M5 can basically degrade 30g / L of xylan in 3-5 days and can be used as the sole carbon source for growth; after the xylan is degraded into xylose, pyruvic acid is obtained by xylose isomerase and xylulokinase; subsequently, acetic acid, ethanol, butyric acid and butanol are obtained through a series of metabolic pathways. According to the strain and the application of the strain disclosed by the invention, enzymes of all pathways can bear a high temperature of 55 to 65 DEG C, and the strain M5 is the one that can produce the butanol by directly utilizing the xylan, which is the only reported bacterial strain until now; therefore, a series of high-temperature resistant enzymes can be provided for industrial production of the butanol, and the strain has important application value.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain that utilizes xylan as the sole carbon source to directly produce butanol. Background technique [0002] As a fuel, butanol has the advantages of high energy density, high stability to water, direct use in internal combustion engines, and convenient transportation. In today's increasingly severe energy crisis, butanol has broad development prospects as a fuel. Butanol is also an important organic chemical raw material, widely used in paints, surface coatings, leather treatment, plastics and other fields. [0003] The production methods of butanol mainly include acetaldehyde condensation method, propylene oxo synthesis method and fermentation method. The acetaldehyde condensation method has a long process flow, low yield and high cost, and has been eliminated abroad at present; the raw material for the production of butanol by the propylene oxo method ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/16C12R1/01
CPCC12P7/16C12R2001/01C12N1/205Y02E50/10
Inventor 信丰学蒋羽佳姜岷董维亮张敏章文明吴昊马江锋
Owner NANJING UNIV OF TECH
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