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Industrialized preparation method of lipopeptide biosurfactant

A bio-surface, active agent technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of low charging coefficient, complex process, long cycle, etc., and achieve high charging coefficient and simple process. , the effect of short cycle

Active Publication Date: 2013-11-27
DAQING HUALI ENERGY BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to defects such as low output, long cycle time, low charging coefficient, and complicated process, the existing surfactants can not be mass-produced.

Method used

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  • Industrialized preparation method of lipopeptide biosurfactant
  • Industrialized preparation method of lipopeptide biosurfactant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Shake flask and fermenter medium formulations are: rice bran oil 2%, urea 1%, molasses 11%, KCl 0.1%, KH2PO4 0.05%, K2HPO4 0.3%, yeast extract 0.005%, compound trace elements 0.008%, the rest is water, The pH is 7.0, and the composite trace elements are zinc sulfate 1.1 g / L, tin oxide 1.3 g / L, manganese chloride 1.0 g / L, copper sulfate 1.0 g / L, cobalt oxide 1.0 g / L and solvent water compounding Matched.

[0027] Shake flask and fermenter culture:

[0028] Inoculate Bacillus subtilis (ACCC01430) into seven 3L shake flasks, fill the shake flasks with 1L medium, insert 2 rings of strains into each shake flask, and cultivate them at 35±2°C and 120 rpm for 14 h. When the number of bacteria detected by the blood cell counting board reaches more than 200 million, the shake flask strain is obtained; then transferred to a 200-liter primary fermentation tank, the filling coefficient is 70%, the inoculation amount is 5%, and the temperature is 35±2°C. The ventilation ratio is 1:...

Embodiment 2

[0030] Shake flask and fermenter medium formulations are: rice bran oil 3%, urea 1.5%, molasses 17%, KCl 1.0%, KH 2 PO 4 0.2%, K 2 HPO 4 1.5%, yeast extract 0.02%, compound trace elements 0.01%, the rest is water, pH is 7.0, the compound trace elements are zinc sulfate 1.1 g / L, tin oxide 1.3 g / L, manganese chloride 1.0 g / L , copper sulfate 1.0 g / L, cobalt oxide 1.0 g / L and solvent water.

[0031] Shake flask and fermenter culture:

[0032] Transfer the slanted solid Bacillus subtilis (ACCC01430) strains into four 3L shaker flasks (each filled with 1L medium), insert 1 ring of strains per 500ml medium, at a temperature of 35±2°C, 120 rpm Cultivate for 13.5 hours, and after the number of bacteria detected by the hemocytometer reaches more than 200 million, transfer it to a 200-liter primary fermentation tank with a filling coefficient of 67%, an inoculum size of 3%, a temperature of 35±2°C, and a ventilation ratio of 1. : 0.4, pH 7.0, stirring speed 120 rpm for 12 hours, tr...

Embodiment 3

[0034]Shake flask and fermenter medium formulations are: rice bran oil 5%, urea 0.2%, molasses 20%, KCl 0.05%, KH 2 PO 4 1.2%, K 2 HPO 4 0.05%, yeast extract 0.1%, compound trace elements 0.005%, the rest is water, pH is 7.5, the compound trace elements are zinc sulfate 1.1 g / L, tin oxide 1.3 g / L, manganese chloride 1.0 g / L , copper sulfate 1.0 g / L, cobalt oxide 1.0 g / L and solvent water.

[0035] Shake flask and fermenter culture:

[0036] Transfer the slanted solid Bacillus subtilis (ACCC01430) strain into six 3L shake flasks (loaded with 1L medium), culture at 35±2°C, 120 rpm for 12h, and the number of bacteria detected by the hemocytometer plate reaches more than 200 million. Connect to a 200-liter primary fermenter with a filling factor of 60%, an inoculum size of 5%, a temperature of 35±2°C, a ventilation ratio of 1:1, a pH of 7.5, and a stirring speed of 120 rpm for 12 h, then transfer Put into a 2-ton secondary fermentation tank, the filling factor is 60%, the ...

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Abstract

The invention provides an industrialized preparation method of a lipopeptide biosurfactant and relates to a production method of a biosurfactant which is prepared from bacillus subtilis ACCC01430 through fermentation. The method comprises the following steps: slope solid bacillus subtilis strains are joined into a shaking bottle for cultivation at the temperature of 35+ / -2 DEG C, and are cultured by shaking at 120 revolutions per minute for 12-16 hours so as to obtain shaking bottle strains; the prepared shaking bottle strains are joined into a primary fermentation tank provided with a culture medium to obtain a primary fermentation strain; the primary fermentation strain is joined into a secondary fermentation tank provided with a culture medium to prepare a secondary fermentation strain; the secondary fermentation strain is joined into a third-level fermentation tank provided with a culture medium to be fermented for 24-26 hours, and then the lipopeptide biosurfactant is obtained. In the industrialized preparation method of the lipopeptide biosurfactant disclosed by the invention, the content of lipopeptid achieves 7-10%g / L, an industrialized production experiment is successful, the fermentation cycle is shortened to 48 hours, and the production cost is greatly reduced. According to the invention, industrially produced lipopenic fermentation liquor is applied on site firstly, and is injected into an underground oil layer, so water is smoothly injected into the original blocks in which water is hard to be injected, the recovery ratio of crude oil is increased by 5-12%, and the efficiency is high.

Description

technical field [0001] The invention relates to a production method of a biosurfactant, in particular to an industrial preparation method of a lipopeptide biosurfactant. Background technique [0002] Lipopeptides, also known as fatty acylpeptides, belong to lipids containing amino acids. Lipopeptide biosurfactants are generally metabolites of Gram-positive Bacillus. Lipopeptide molecules are composed of two parts: hydrophilic peptide bonds and lipophilic aliphatic hydrocarbon chains. Due to their special chemical composition and amphiphilic molecular structure, lipopeptide biosurfactants are widely used in the fields of medicine, food, cosmetics and microbial oil recovery. There are important application prospects. Lipopeptide surfactants are generally metabolized by Gram-positive Bacillus. There are many strains that can produce lipopeptide surfactants, such as Bacillus subtilis and Bacillus licheniformis. In addition to bacteria that can produce lipopeptide surfactants,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00C12R1/125
Inventor 沈玉江孙彦达李国军王柱张丽茹段宝颜
Owner DAQING HUALI ENERGY BIOLOGICAL TECH
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