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Novel seawater fermentation strain generating biosurfactant

A surfactant and biological surface technology, applied in the field of fermentation engineering, can solve problems such as harm to health, unreasonable recycling, environmental pollution, etc.

Inactive Publication Date: 2014-10-08
YANCHENG TEACHERS UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of the traditional fermentation water is fresh water, and the cost of seawater desalination is relatively high at present. Therefore, it is of special significance to use new seawater fermentation strains to produce biosurfactant rhamnolipids in coastal tidal areas where fresh water is scarce and the water quality is poor.
[0004] The cost of fermented carbon source raw materials is also an important part of the production cost of fermentation engineering, but waste edible oils such as gutter oil and frying oil have not been well utilized at present Instead, it becomes a serious environmental pollutant or is unreasonably recycled and returned to the dining table, endangering people's health

Method used

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  • Novel seawater fermentation strain generating biosurfactant
  • Novel seawater fermentation strain generating biosurfactant
  • Novel seawater fermentation strain generating biosurfactant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: Screening and isolation steps of novel biosurfactant seawater fermentation strains

[0020] (1) Sampling

[0021] The following samples are mainly collected: Soil or mud samples are collected from coastal tidal flats. Water samples were collected from offshore seawater near the port. Bring back immediately after collection and store at 4°C.

[0022] (2) Enrichment culture

[0023] Add soil samples or mud samples to shake flasks with seawater fermentation medium, shake culture at 30°C for 3 days, then transfer to new seawater fermentation medium, enrich and cultivate for 3 times, so that waste edible oil can be used as the only Carbon source microorganisms are enriched.

[0024] (3) Primary screening

[0025] Take the enrichment solution in (2) and streak or dilute the culture on the high-salt blood plate, pick a single colony with a large and clear transparent circle and connect it to the slope at 30°C for 1-2 days, then store it at 4°C for later use....

Embodiment 2

[0036] Embodiment 2: Identification of seawater fermentation strain Z-2

[0037] The physiological and biochemical identification and 16SrDNA molecular identification of the seawater fermentation strain Z-2 were carried out, and the species of the seawater fermentation strain was determined by combining the morphological characteristics and physiological and biochemical characteristics of the bacteria.

[0038] (1) Cell morphology, physiological and biochemical characteristics of seawater fermentation strain Z-2

[0039] According to the "Common Bacterial System Identification Handbook", including morphological microscopy, spore production, Gram staining, pyocyanin, indole production, nitrate reduction, glucose oxidative fermentation, catalase, gelatin liquefaction, starch hydrolysis, Citrate, methyl red, acetyl methyl carbinol (V-P), malonate and other experiments. Determination result is as follows table 1:

[0040] Table 1 Cell morphology and physiological and biochemical...

Embodiment 3

[0047] Embodiment 3: Determination of seawater fermentation strain Z-2

[0048] The 16SrDNA sequencing results of the strain were homologously compared with the sequences in GenBank, and compared by BLAST, it had a high homology with Pseudomonas sp. Retrieve relevant 16SrDNA sequences from GenBank, use software such as ClustalX and Mega5 for phylogenetic analysis, and use neighbor-joining method to construct phylogenetic tree ( image 3 ). Based on the results of physiological and biochemical experiments (Table 1) and molecular identification and cluster analysis of this strain, it can be determined that the seawater fermentation strain Z-2 is Pseudomonas aeruginosa. Its classification is named Pseudomonas aeruginosa (pseudomonas aeruginosa), and it has been preserved in the General Microbiology Center of the China Committee for the Collection of Microorganisms (Address: Institute of Microbiology, Chinese Academy of Sciences, No. 1, Beichen West Road, Chaoyang District, Beiji...

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Abstract

Traditional fermentation generally adopts a freshwater resource. The invention discloses a novel seawater fermentation strain generating a biosurfactant. Fermentation with natural seawater as fermentation water and waste edible grease as a single carbon source is carried out to produce the biosurfactant rhamnolipid. Seawater fermentation enterprises can be built in coastal regions lacking a freshwater resource and having bad water quality to solve the dependence of the traditional fermentation industry on the freshwater resource. The strain is separated and purified through sweater enrichment culture, hypersalemia flat primary screening, synthetic medium secondary screening and seawater shaking bottle fermentation verification, and is determined as Pseudomonas aeruginosa through physiologic biochemical experiments and molecular identification analysis, is preserved in China General Microbiological Culture Collection Center on April 25, 2014, and has a preservation number of CGMCC No.9091. The strain uses the advantages of the seawater fermentation, and the obtained crude product rhamnolipid can be used in regional market promotion, and has a potential application prospect in the fields of the agriculture, the petroleum chemical industry, environment treatment, soil restoration, the cosmetic industry, the pharmacy industry and the food industry.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, and in particular relates to a seawater fermentation type bacterial strain using natural seawater as fermentation water and waste edible oil as a single carbon source to ferment and produce biosurfactant rhamnolipid. Background technique [0002] Most of the surfactants currently on the market are mainly derived from petroleum-based chemical products. The production process requires a large amount of petroleum products and is easy to cause environmental pollution. However, the acquisition of biosurfactants does not have such problems. Biosurfactant rhamnolipid is a new type of anionic biosurfactant obtained from the secondary metabolism of microbial fermentation. It has the characteristics of biodegradability, low toxicity and environmental friendliness. And because rhamnolipid has good emulsification, wettability, and foaming properties, it has potential application prospects in agricultu...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/44C12R1/385
Inventor 张祥胜张飞苗丽平周冲
Owner YANCHENG TEACHERS UNIV
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