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Method for extracting rhamnolipid product in fermentation liquid

A technology of rhamnolipid and fermentation broth, which is applied to the preparation of sugar derivatives, sugar derivatives, sugar derivatives, etc., can solve the problems of high toxicity, high cost, etc., and achieve the effect of quick extraction and avoidance of use.

Active Publication Date: 2018-06-22
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The high cost and high toxicity of the above extraction methods limit the fermentative production and application of rhamnolipids

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Extraction of rhamnolipid products from fermentation broth with glucose as carbon source.

[0021] 1) Preparation of fermentation medium: Weigh separately with an electronic balance, 30g glucose, 3g NaNO 3 , 1g KH 2 PO 4 , 2gK 2 HPO 4 ·3H 2 O, 0.4g MgSO 4 ·7H 2 O, 1g of yeast powder; add 200ml of distilled water and mix well, pour it into a volumetric flask, and dilute to 1L with distilled water; add NaOH with a mass fraction of 30%, and adjust the pH of the medium to 7.0; divide it into a triangular flask, place Sterilize at 115°C for 30 minutes.

[0022] 2) Seed solution preparation: the slant strain 2 inoculation loop of the rhamnolipid-producing Pseudomonas aeruginosa (Pseudomonas aeruginosa) ATCC27853 was inoculated into the Erlenmeyer flask containing 100ml LB medium, at 37°C and 180rpm / min , cultured for 6-10 hours, to OD 600 The value is 0.7, that is, the seed solution cultivated to the late logarithmic growth period is obtained, and the numb...

Embodiment 2

[0027] Embodiment 2: selected bacterial strain, culture medium composition, culture condition are identical with embodiment 1, difference is that used chitosan has different deacetylation degree, it is respectively set to add deacetylation degree to be 85%, 90% and 92 % experimental treatment of 3 kinds of chitosan, using the method of the present invention to extract the rhamnolipid in the fermentation broth. The yields of rhamnolipids were 45.8%, 60.8% and 62.1%, respectively; the results showed that when the deacetylation degree of chitosan was ≥90%, the yield of rhamnolipid products was ≥60%.

Embodiment 3

[0028] Embodiment 3: Extract rhamnolipid product from the fermented liquid taking soybean oil as carbon source

[0029] Prepare fermentation medium with soybean oil as carbon source, weigh with electronic balance, 3g NaNO 3 , 1g KH 2 PO 4 , 2g K 2 HPO 4 ·3H 2 O, 0.4g MgSO 4 ·7H 2 O, 1g yeast powder; add 200ml of distilled water and mix well, pour it into a volumetric flask, and use distilled water to make up to 1L; add NaOH with a mass fraction of 30%, and adjust the pH of the medium to 7.0; The concentration is 30g / L, add soybean oil into the Erlenmeyer flask respectively; then sterilize at 121°C for 20min.

[0030] Rhamnolipid-producing Pseudomonas aeruginosa (Pseudomonas aeruginosa) DSM 22644 slant strain 2 inoculation loop, inoculated into the Erlenmeyer flask containing 100ml LB medium, at 37 ℃, 180rpm / min, culture 6-10 hours to OD 600 The value is 0.7, that is, the seed solution cultivated to the late logarithmic growth period is obtained, and the number of bact...

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Abstract

The invention discloses a method for extracting a rhamnolipid product in fermentation liquid. The method comprises the following steps of (1) pretreatment: heating fermentation liquid; after cooling,removing insoluble substances such as thalli and denatured protein through centrifugation; (2) flocculation and concentration: adding proper amount of chitosan into supernate; performing uniform mixing; performing natural settling at proper temperature; removing liquid phases at the upper layer; (3) elution: removing residual fermentation liquid in the precipitate through cleaning by distilled water; then, adding a NaHCO3 solution with certain proper concentration; eluting rhamnolipid in the precipitate; collecting elution liquid; (4) obtaining: regulating the elution liquid pH; performing still settlement and centrifugation to obtain the rhamnolipid product. The method is a simple and convenient method for efficiently extracting rhamnolipid in the fermentation liquid; important application values are realized in the field of rhamnolipid fermentation production.

Description

technical field [0001] The invention belongs to the field of rhamnolipid fermentation production, in particular, it relates to a method for extracting rhamnolipid products in fermentation broth. Background technique [0002] Rhamnolipid is a biosurfactant synthesized by the metabolism of bacteria such as Pseudomonas aeruginosa. Because of its high surface activity, low critical micelle concentration, safety, non-toxicity, and biodegradability, it is widely used in microbial oil recovery. , Environmental pollution remediation, food, biomedicine and other technical fields have broad application prospects. [0003] The fermentation broth of rhamnolipid-producing bacteria has complex components, and the cost and difficulty of extracting rhamnolipids from the fermentation broth are high. At present, the methods for extracting rhamnolipids from fermentation broth are mainly chloroform-methanol extraction, silica gel column chromatography, etc., and a large amount of chloroform ne...

Claims

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Application Information

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IPC IPC(8): C07H15/04C07H1/08B01F17/56C09K23/56
CPCC07H1/08C07H15/04C09K23/00
Inventor 赵峰张颖史荣久韩斯琴
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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