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Method for producing fatty acid by mixing and fermenting oil-producing yeast and bacillus to degrade lignocellulose

A technology of lignocellulose and bacillus, which is applied in the field of mixed fermentation and degradation of lignocellulose to produce fatty acid by oleaginous yeast and bacillus, can solve the problems of high cost and low economic feasibility.

Inactive Publication Date: 2015-02-25
ZHENJIANG BIO INNOVA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For most microorganisms, the most suitable carbon source for growth is generally glucose, so there have been many studies on oleaginous yeast using glucose as a carbon source, and the highest oil content in the bacteria can reach 80%, but The cost of large-scale microbial oil production using glucose is high and the economic feasibility is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] seed pot culture

[0009] The used seed tank culture medium formula of the present embodiment is as follows:

[0010] A 5L fermenter was used as a seed tank to prepare a 3L fermentation broth.

[0011] Glucose 10g / L, yeast extract 5g / L, urea 1g / L, (NH 4 ) 2 SO 4 1.5g / L,

[0012] MgSO 4 ·7H 2 O 1.5g / L, Na 2 HPO4·12H 2 O 9g / L, KH 2 PO4 1.5g / L. Except for glucose, other components of the medium were sterilized at 121°C for 20 minutes, and glucose was sterilized at 115°C for 20 minutes alone. The inoculation volume is 10%,

[0013] Oleaginous yeast culture conditions: initial pH 7.0±0.02, 30°C, 200rpm, cultured for 24h.

[0014] Bacillus culture conditions: initial pH 9.0±0.02, 30°C, 200rpm, culture for 24h.

[0015] Expand training

[0016] The contents of each component of the fermentation medium used in the present embodiment are as follows:

[0017] Lignocellulose 15g / L, urea 1g / L, MgSO 4 ·7H 2 O 1g / L, CaCl 2 2H 2 O 0.02g / L, FeSO 4 ·7H 2 O 0.5g / L,...

Embodiment 2

[0024] Example 2 (determination of dry weight of oleaginous yeast cells)

[0025] Take 1ml of the fermentation broth after the completion of the above-mentioned Example 2, centrifuge at 10000rpm for 10min, wash the biomass precipitate twice with water, and dry at a constant temperature at 40°C for 24h, and determine the biomass by gravimetric analysis.

Embodiment 3

[0026] Example 3 (extraction of oil in yeast cells by acid heat method)

[0027] Get the dry thalline 1g that above-mentioned embodiment 3 makes, add 10mL concentration and be the hydrochloric acid of 4mol / L

[0028] Shake and mix

[0029] Stand for 1 hour at 20°C

[0030] Boiling water bath 8-10min

[0031] Cool at room temperature for 30 minutes

[0032] The solution was added to the separatory funnel, and 5ml of absolute ethanol was added

[0033] Add 25ml chloroform:methanol (2:1) mixture to extract yeast intracellular oil

[0034] The organic layer of the lower phase was collected, and the lower phase containing lipid was evaporated in a rotary evaporator under a nitrogen flow atmosphere, and the fat was recovered and weighed.

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PUM

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Abstract

The invention discloses a method for producing fatty acid by mixing and fermenting oil-producing yeast and bacillus to degrade lignocellulose. The method comprises the following steps: grinding straws or wood chips into fine granules, then treating the ground lignocellulose raw materials with dilute alkali, so as to be thoroughly dissolved; centrifuging cultured bacillus and washing twice with normal saline so as to entirely removal carbon source compositions, transferring to a lignocellulose-containing fermentation culture medium for culturing for 96-144h; inoculating the oil-producing yeast to the fermentation culture medium for continuing fermentation for 96-144h after culturing for 96h; after the culturing is over, centrifuging and collecting cells, breaking walls to extract biolipid. The method has the advantages that compared with other technologies, microbial lipid can be produced under low energy consumption through the research, and the sources of raw materials can be increased.

Description

technical field [0001] The invention relates to a method for microbial degradation of lignocellulose to produce fatty acid, in particular to a method for mixed fermentation of oleaginous yeast and bacillus to degrade lignocellulose to produce fatty acid. Background technique [0002] Lignocellulose is a complex composed of a variety of complex polymer organic compounds, mainly composed of three main aggregates of cellulose, hemicellulose and lignin, and some extracts soluble in polar or weakly polar solvents. Lignin and hemicellulose are covalently bonded to surround cellulose to form a strong natural barrier, so lignocellulose is difficult to degrade during direct hydrolysis. In addition, harmful inhibitors to fermentation and subsequent hydrolysis will be produced during the hydrolysis of lignin, and cellulose itself has a complex crystal structure, so the effect of direct hydrolysis of lignocellulose is poor, and it can degrade lignocellulose. Microorganisms are mainly f...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P7/64C12R1/07C12R1/645
Inventor 朱道辰谢长校陈焱
Owner ZHENJIANG BIO INNOVA BIOTECH
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