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Method and application of designing enzyme preparation feeding mode in fermentation process by computer

A technology of fermented raw materials and fermented liquid, which is applied in the fields of renewable energy and bioengineering, can solve problems such as the difficulty in the rational design of cellulase formulations, and achieve the effects of saving production costs, reducing additives, and improving economic efficiency

Active Publication Date: 2022-01-18
NANJING NORMAL UNIVERSITY
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Problems solved by technology

[0004] The purpose of the present invention is to overcome the problem of difficulty in the rational design of cellulase formulations in the prior art, and to provide a method and application of computer-designed enzyme preparation feeding methods in the fermentation process, which can optimize fermentation to produce ethanol Timely cellulase addition strategy can increase ethanol production and reduce costs, thereby improving the economic benefits of the entire ethanol process

Method used

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  • Method and application of designing enzyme preparation feeding mode in fermentation process by computer
  • Method and application of designing enzyme preparation feeding mode in fermentation process by computer
  • Method and application of designing enzyme preparation feeding mode in fermentation process by computer

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preparation example Construction

[0064] The second aspect of the invention provides a method of producing biohal ethanol comprising: forming a fermentable raw material by liquefied treatment, inoculation of ethanol fermentation strain to the fermentation raw material, and adds a glycosidase, nitrogen source Synchronized saccharification fermentation is performed, and the enzyme preparation feed during the fermentation process obtained according to the above method is added, and the respective enzyme component is added to the feed period.

[0065]By activity of cellulase components measured at different simulated concentrations seen in all cellulase - ethanol system, the ethanol concentration is between 0-16% (v / v), gradually increasing the number of ethanol molecules, more significantly inhibited cellulase will occur with increasing concentration of ethanol, cellulase activity decreases, which certainly limits the efficient hydrolysis of starch the cellulose in the feedstock to glucose, thereby interfering with...

Embodiment 1

[0078] (1) Measurement of the activity of exo-cellulase I (CBHI-Tr): cutting the outer 0.1mL I cellulase solution, 0.7mL concentration of 10mmol / L of 4-nitrophenyl fibers -β-D- diglycoside 0.1mL solution at a concentration of 0.05M citrate buffer, mixed, and ethanol was added to obtain a mixture, such a mixture of ethanol concentration of 0 vol%, 4 vol%, 8 vol%, 12 vol%, 16 vol %, 20 vol%, 40min followed by reaction at 50 ℃, 2mL was added at a concentration of 1mol / L of Na 2 CO 3 10mL water and, measuring absorbance at 400nm, the results shown in Table 1;

[0079] (2) Measurement of the activity of exo-cellulase II (CBHII-Hi): 200-fold diluted to 0.1mL exocellulases II solution, 0.14mL concentration of 2 wt% solution of microcrystalline cellulose PH101, 0.1mL 0.05M citrate buffer at a concentration of mixed solution, and ethanol was added to obtain a mixture, such that the ethanol concentration in the mixture was 0% by volume, respectively, 4% by volume, 8 vol%, 12 vol%, 16 vo...

Embodiment 2

[0087] Culturing 24h (1) to add a YPD medium with glucose as a seed medium Saccharomyces cerevisiae (glucose concentration of 100g / L), at a temperature of 30 ℃, speed of 150rpm conditions to give a seed culture broth, the seeds the culture was centrifuged at 4000rpm 10min seed obtained wet cells;

[0088] (2) The corn flour was mixed with water such that the dry matter concentration of the corn is 30 wt%, 0.012 wt.% Alpha] amylase (relative to the dry corn weight), after the temperature of the liquefied 90min under conditions of 88 deg.] C, cooled to 60 ℃ fermentation feedstock obtained;

[0089] (3) 30g of step (2) as a raw material obtained in the fermentation broth, the step (1) seed obtained wet cells was inoculated to the dry cell weight 0.44g / kg broth, was added 0.0325 wt% (with respect to the maize dry weight) of glucoamylase and 0.012 wt% (relative to the dry weight of corn) urea, then the value of pH 4.0, rotation speed of 150 rpm, the fermentation temperature is at 3...

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Abstract

The invention relates to renewable energy and bioengineering technology, and discloses a computer-designed enzyme preparation feeding method in the fermentation process and its application. The method comprises: under the condition that the target product of the fermentation process exists, respectively measuring the activity of each enzyme component, wherein the concentration of the target product is set to a plurality of different simulated concentrations; obtaining the crystal structure of the enzyme component from a protein database, Select the model of the target product, and carry out the molecular dynamics simulation of the enzyme component when the concentration of the target product is the simulated concentration; obtain the concentration value of the target product during the fermentation process without adding the enzyme preparation, and compare the activity data with the molecular dynamics simulation results In combination, the feeding time period and feeding amount of each enzyme component were proposed. The method can optimize the adding strategy of cellulase during fermentation to produce ethanol, increase the yield of ethanol, reduce the cost, and further improve the economic benefit of the whole ethanol process.

Description

Technical field [0001] The present invention relates to renewable energy and biological engineering techniques, and more particularly to methods and applications of enzyme preparations during fermentation by a computer. Background technique [0002] Biological ethanol is a very promising liquid fuel that meets the growing energy demand and sustainable economy, and has a positive impact on rural, environmental and energy security. A large number of research work on biological ethanol has been launched in recent years, and advanced fermentation preparation techniques have been developed to improve the economics of the entire biological ethanol process and increase profitability. However, the industry still urgently requires higher biomass utilization and ethanol production. [0003] During the production of biological ethanol, hydrolysis is a key step of converting cellulose biomass (corn, corn stalk, wheat straw, willow branches, etc.) into ethanol, except for chemicals (mainly ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/06C12Q3/00G16C10/00
CPCC12P7/06C12Q3/00G16C10/00Y02E50/10Y02P20/59
Inventor 黄和李秀娟乔杰王明慧盛义杰张立慧魏珺楠
Owner NANJING NORMAL UNIVERSITY
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