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Bacillus amyloliquefaciens for producing surfactin, and application thereof

A technology of amylolytic spores and surfactin, which is applied in the field of microorganisms, can solve problems such as the lack of amyloliquefaciens, and achieve the effects of high surface activity, high-efficiency crude oil and guar gum degradation ability, and high crude oil emulsification and viscosity-reducing ability

Active Publication Date: 2016-10-12
大连知微生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are few relevant reports on the application of Bacillus amyloliquefaciens and its biologically active products in the field of microbial oil recovery and environmental remediation

Method used

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  • Bacillus amyloliquefaciens for producing surfactin, and application thereof
  • Bacillus amyloliquefaciens for producing surfactin, and application thereof
  • Bacillus amyloliquefaciens for producing surfactin, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Screening and Identification of Bacterial Strain CB-019

[0039] 1. Isolation of strain CB-019

[0040] (1) Sample source: produced fluid from an oilfield.

[0041] (2) Preliminary screening of bacterial strains: dilute the sample with sterile water to 10 -3 ~10 -8 , take 0.1mL and spread evenly on the blood plate medium, and put it in a 37°C incubator for cultivation. Transfer the colony with the hemolytic circle to a fresh blood plate, streak and purify it, inoculate it into the slant medium, store it at 4°C, and keep it for later use.

[0042] (3) Re-screening of strains: inoculate the preserved strains into LB medium, shake and culture at 37°C and 160rpm for 24 hours, then inoculate into the fermentation medium with an inoculum size of 1:50, and culture with shake at 160rpm at 37°C for 30 hours. ~36h. Take 30 mL of the above fermentation broth, centrifuge at 5000 r / min and measure the surface tension of the supernatant. Among them, the strain with th...

Embodiment 2

[0048] Example 2 Extraction and Identification of Fermentation Products of Bacterial Strain CB-019

[0049] Fermentation of strain CB-019: Inoculate Bacillus amyloliquefaciens CB-019 on LB solid medium, culture at 35°C for 1d, inoculate a single colony in the seed medium, and culture with shaking at 35°C and 150rpm for 2d , to become a seed solution, which was added to the fermentation medium at a volume ratio of 1:20, shaken and cultured at 35°C and 150 rpm for 3 days, and centrifuged to obtain a fermentation solution.

[0050] Extract the fermentation product in CB-019 from the fermentation broth by acid precipitation method: centrifuge the fermentation broth to remove the bacteria, adjust the pH of the supernatant to 2.0, let it stand overnight, collect the precipitate by centrifugation, wash the precipitate with hydrochloric acid at pH 2.0, and wash it with NaHCO 3 Adjust the hydrochloric acid washing liquid to pH 8.0, add chloroform methanol (3:1) mixture as the extractan...

Embodiment 3

[0054] Example 3 Determination of the performance of bacterial strain CB-019 biologically active substances

[0055] The fermentation broth of the bacterial strain CB-019 obtained in Example 2 was used to perform the following determinations.

[0056] 1. Determination of emulsifying performance

[0057] (1) Determination of the oil discharge circle of the fermentation broth: 0.5 g of Sudan Red III was added to 100 mL of liquid paraffin, the liquid paraffin was dyed, filtered and set aside. Add 8mL of dyed liquid paraffin to a 9cm plate containing 60mL of distilled water. After a layer of uniformly dispersed dyed liquid paraffin film is formed on the surface of the plate, slowly drop 1mL of the fermentation broth of strain CB-019 into the center of the oil film, and observe The size of the oil drain ring, the result is as follows Figure 5 As shown, the results show that CB-019 can produce a larger oil discharge ring, with a diameter of about 5-6cm.

[0058] (2) Emulsificati...

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Abstract

The invention discloses a Bacillus amyloliquefaciens CB-019 for producing surfactin. The above strain is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No.11950. A method for fermenting the strain and preparing a biochemical oil displacement agent is simple and is easy to implement, and the prepared biochemical oil displacement agent has good environment adaptability, and can keep high surface interface vitality and crude oil emulsification viscosity reduction ability; and the strain also can degrade petroleum, guar gum and other high-molecular polymers, and can be applied to treatment of oil-containing sewage, backflow fracturing fluids and polymer-bearing wastewater. The Bacillus amyloliquefaciens CB-019 has important promotion and application values in the fields of microbial oil extraction and environment restoration.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a strain of bacillus amyloliquefaciens producing surfactin and its application in the fields of microbial oil recovery and environmental restoration. Background technique [0002] Biosurfactants are surface-active substances metabolized by microorganisms under specific conditions. They can be divided into glycolipids, lipopeptides and lipoproteins, fatty acids and phospholipids, polymers and the surface of whole cells according to their chemical composition and microbial sources. five categories. Like chemically synthesized surfactants, the molecular structure of biosurfactants also consists of two parts, an oleophobic and hydrophilic polar group and a hydrophobic and lipophilic non-polar group. However, compared with chemical surfactants, biosurfactants have their own powerful advantages, such as wide selectivity, environmental friendliness, various types of molecular stru...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C09K8/582C09K8/584C12R1/07C02F101/32C02F101/34
CPCC02F3/34C02F3/344C02F2101/32C02F2101/34C09K8/582C09K8/584C12N1/205C12R2001/07
Inventor 赵静韩俊杰
Owner 大连知微生物科技有限公司
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