A high-efficiency xylose-fermenting mutant strain and a method for producing ethanol by fermentation thereof

A technology for ethanol production and mutant strains, which is applied in the field of high-solid lignocellulose ethanol fermentation, and can solve problems such as temperature tolerance that needs to be improved

Active Publication Date: 2019-06-14
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it still needs to be improved in terms of temperature tolerance and ethanol production performance from xylose fermentation

Method used

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  • A high-efficiency xylose-fermenting mutant strain and a method for producing ethanol by fermentation thereof
  • A high-efficiency xylose-fermenting mutant strain and a method for producing ethanol by fermentation thereof
  • A high-efficiency xylose-fermenting mutant strain and a method for producing ethanol by fermentation thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: U1-58 mutant strain mutagenesis screening

[0036] (1) Mutagenesis and primary screening S. passalidarum NRRL Y-27907 was used as the starting strain (hereinafter referred to as U1 strain), plasma mutagenesis was carried out for 25s, and the mutagenized bacterial solution was diluted to an appropriate concentration, and spread on the primary screening plate Above, after 1-2 days of inverted culture at 37°C, mutants with better growth and smooth surface were picked for re-screening.

[0037] Primary screening plate: xylose 50g / L, peptone 20g / L, yeast extract powder 10g / L, agar 20g / L, pH natural.

[0038] (2), double screening carries out xylose culture medium shake flask fermentation double screening to the mutant strain gained in (1), fermentation medium is: xylose 100g / L, yeast extract powder 22g / L, K 2 HPO 4 2.5g / L, MgSO 4 0.1g / L.

[0039] The fermentation conditions are: liquid volume 100mL / 250mL, strain initial OD 600 1.0, pH 5.5, temperature 3...

Embodiment 2

[0041] Example 2: Determination of the growth performance of the U1-58 mutant strain and the U1 starting strain at different temperatures

[0042] The U1-58 mutant strain and the U1 starting strain were used as mutual controls, and the growth curves of the strains were measured at 30°C, 33°C, 35°C, 37°C, 39°C and 41°C.

[0043] Determination method: Pick one ring of bacteria from the inclined plane and put it into a 250mL shaker flask filled with 50mL YPX liquid medium (20g / L xylose, 20g / L peptone, 10g / L yeast extract powder), at 30°C and 180rpm Cultivate, take samples at regular intervals, dilute appropriately, and measure the OD value at 600nm. Measurement results such as figure 2 shown.

[0044] from figure 2 It can be clearly seen that the temperature is between 30°C and 41°C. Although the growth of the strain is inhibited to varying degrees as the temperature increases, it can still be clearly seen that the growth performance of the U1-58 mutant strain is similar to ...

Embodiment 3

[0045] Example 3: Determination of xylose fermentation performance of U1-58 mutant strain at different temperatures

[0046] Using the U1-58 mutant strain and the U1 starting strain as mutual controls, the xylose fermentation performance of the strains was measured at 30°C, 33°C, 35°C, 37°C, 39°C and 41°C.

[0047] Fermentation condition and the mensuration of metabolite concentration in fermented liquid are all the same as embodiment 1 (2). The residual xylose concentration, ethanol concentration and corresponding sugar alcohol conversion rate in the fermented liquid after fermentation are analyzed as indicators, and the fermentation results are as follows: image 3 shown.

[0048] like image 3 As shown, at each temperature, the U1-58 mutant strain had a significantly higher ethanol concentration and a significantly lower residual sugar concentration than the U1 starting strain, and the corresponding sugar alcohol conversion rate also increased to a certain extent. At 30°...

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Abstract

The invention belongs to the technical field of microbial fermentation, and in particular relates to a mutant strain of Spathaspora passalidarum obtained through mutagenesis and screening that can efficiently ferment xylose to produce ethanol and a method for using it to ferment high-solid lignocellulose ethanol. The mutant strain overcomes the problems of low ethanol tolerance, difficult high-solids fermentation, and poor ethanol production ability of using xylose in the prior art, and has high temperature tolerance, and can ferment xylose to produce ethanol at 33°C. The concentration reaches 44.19g / L, and the co-fermentation of glucose and xylose by lignocellulose hydrolyzate can reach an ethanol concentration of 43.26g / L, and the synchronous saccharification and co-fermentation of high-solid lignocellulose can reach an ethanol concentration of 49.92g / L. Significant progress has been made over the prior art.

Description

Technical field: [0001] The invention belongs to the technical field of microbial fermentation and the technical field of biotransformation of fibrous substances, and specifically relates to a mutant strain of Spathaspora passalidarum U1-58 obtained through mutagenesis and screening that can efficiently ferment xylose to produce ethanol, and the U1-58 mutant strain for high Method for ethanol fermentation of solid lignocellulose. Background technique: [0002] Energy and environmental protection are two big mountains restricting my country's economic development. Finding sustainable, renewable and clean energy has become an important development strategy for our country. Ethanol is the main form of biomass liquid energy, clean and renewable, and an ideal substitute for fossil fuels. More importantly, ethanol can also provide raw materials for some chemical raw materials such as ethylene. Lignocellulose is considered to be the most potential raw material for ethanol product...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P7/06C12R1/645
CPCC12P7/06C12N1/145C12R2001/645Y02E50/10
Inventor 陈叶福肖冬光付更新李保忠郭学武
Owner TIANJIN UNIV OF SCI & TECH
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