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In situ detoxication alcohol fermentation method of ligno-cellulose hydrolysate using single strain

A lignocellulose and ethanol fermentation technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems such as the complexity of the ethanol fermentation process and increase the cost, and achieve the effect of low production cost

Inactive Publication Date: 2010-07-07
CAPITAL NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hydrolyzate is usually detoxified before fermentation, which requires the addition of treatment equipment, and the detoxification equipment must be well integrated with the fermentation process, thus increasing the complexity of the entire ethanol fermentation process and increasing the production cost [7]

Method used

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  • In situ detoxication alcohol fermentation method of ligno-cellulose hydrolysate using single strain
  • In situ detoxication alcohol fermentation method of ligno-cellulose hydrolysate using single strain
  • In situ detoxication alcohol fermentation method of ligno-cellulose hydrolysate using single strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The separation and identification of embodiment 1 Y7

[0024] 1 Screening process

[0025] It was isolated from the sewage sludge of the alcohol factory in our laboratory. It was enriched and cultured in the YPD liquid medium added with furfural and hydroxymethylfurfural, and then spread on a solid plate containing xylose to isolate a single colony. After a large number of screenings, it was found that one of the strains had a strong metabolism, could efficiently utilize glucose and xylose in the hydrolyzate, and could tolerate high concentrations of inhibitors. It was named Y7 and was finally preserved on a slant.

[0026] 2 Morphological characteristics

[0027] The colonies are smooth, spherical and small, and reproduce by binary fission without false filaments.

[0028] 3 culture characteristics

[0029] After being cultured in the liquid medium for a period of time, the bacterial solution was turbid without flocculation.

[0030] 4 metabolic characteristics

...

Embodiment 2

[0032] Example 2 Identification of Y7 on the in-situ detoxification ethanol fermentation of lignocellulose dilute acid hydrolyzate

[0033] 1 Materials and methods

[0034] 1.1 Strains

[0035] Strain Pichia.Stipitis Y7

[0036] 1.2 Medium (g / L)

[0037] YPD medium: glucose 20, yeast extract 10, peptone 20, pH 5.0-5.5.

[0038] Fermentation medium: yeast extract 10, peptone 20. In addition, glucose and xylose were added according to different needs (concentration is subject to actual measurement) pH 5.0-5.5.

[0039] 1.3 Dilute acid hydrolyzate of lignocellulose (g / L)

[0040] The dilute acid hydrolyzate of lignocellulose used in this study was provided by East China University of Science and Technology, in which glucose was 44.678, xylose was 24.35, furfural was 1.37, 5-HMF0.47, acetic acid was 10, Ca(OH) 2 Adjust the pH of the hydrolyzate to 5.0.

[0041] 1.4 Culture of strains

[0042] The strain Y7 was inoculated in 100ml of YPD medium and cultured at 30°C and 150r...

Embodiment 3

[0068] Example 3 In situ detoxification ethanol fermentation of lignocellulose dilute acid hydrolyzate by Y7

[0069] 1 seed solution preparation

[0070] Add 100mL YPD liquid medium (glucose 20g / L, peptone 20g / L, yeast extract 10g / L) into a 250mL Erlenmeyer flask, put it into a sterilizing pot at 115°C for 25 minutes for sterilization. Pick a ring of single colony into the Erlenmeyer flask, 150rpm, shaker at 30°C for 12 hours.

[0071] 2 Treatment of hydrolyzate

[0072] Add peptone (20g / L), yeast extract (10g / L), Ca(OH) to the hydrolyzate 2 Adjust the pH to 5.0, put it into a sterilizing pot at 115°C for 25 minutes to sterilize.

[0073] 3 Fermentation of lignocellulose dilute acid hydrolyzate

[0074] Take 10mL Y7 seed solution (the number of viable bacteria is 8×10 8 pcs / ml~9×10 8 Each / ml) was added to the Erlenmeyer flask equipped with hydrolyzate, 150rpm, shaker and cultured at 30°C.

[0075] 4 analysis method

[0076] 4.1 Determination of sugar concentration

...

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Abstract

The invention provides a method for adopting single strains to carry out fermentation to in-situ virus-eliminated alcohol of hydrolyzate of lignocellulose; therefore, the invention firstly provides a new strain of pichia stipitis of trunks and the preservation number is CGMCC No.2661; the experiment proves that the strain can carry out detoxification in situ to lignocellulose diluted acid hydrolyzate, can transform glucose and xylose in lignocellulose diluted acid hydrolysate into ethanol effectively and reach 92.4 percent of the highest theoretical value of the ethanol. The use of the straincan simplify the technique that the lignocellulose is used as materials to produce the ethanol, reduces the production cost of the ethanol and has significant theoretical and practical significance to the commercialization of the production of lignocellulose ethanol.

Description

technical field [0001] The present invention relates to an ethanol fermentation method, specifically an in-situ detoxification ethanol fermentation method for lignocellulose hydrolyzate by using a single strain, and also relates to a new microbial strain for ethanol fermentation, specifically It is a new strain of Pichia.Stipitis. Background technique [0002] In the field of research and development of ethanol production from lignocellulose, dilute acid hydrolysis of lignocellulose is considered to be the easiest production process to achieve commercialization [1] , a large number of in-depth researches have been carried out in this field at home and abroad. But so far, there are still bottlenecks in dilute acid hydrolysis of lignocellulose to produce fuel ethanol at home and abroad. To solve the technical bottleneck is to obtain ethanol from xylose in the hydrolyzed sugar solution by efficient fermentation and efficient metabolism of xylose in the hydrolyzate. Fermentati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P7/10C12R1/84
CPCY02E50/16Y02E50/10
Inventor 杨秀山
Owner CAPITAL NORMAL UNIVERSITY
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