Sphingomonas sp. T-3 and method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation

A sphingomonas, biopolysaccharide technology, applied in the field of biotechnology and biomaterials, can solve the problem of limiting the application of gellan gum and dieter gum, affecting the properties of gellan gum and dieter gum, poly-beta hydroxybutyric acid Problems such as low output, to achieve the effect of good shear thinning performance and temperature resistance, low cost and good industrial value

Active Publication Date: 2015-05-27
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main reason is: firstly, the yield of poly-β-hydroxybutyric acid in these two strains is low; secondly, the separation of gellan gum and Dieter gum from poly-β-hydroxybutyric acid needs to be diluted, the process of membrane separation, and dilution after separation The extraction of gellan gum and Dieter gum in the ferme

Method used

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  • Sphingomonas sp. T-3 and method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation
  • Sphingomonas sp. T-3 and method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation
  • Sphingomonas sp. T-3 and method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Screening and identification of Sphingomonas sp. T-3

[0034] The strain was isolated from oil-contaminated soil.

[0035] The specific implementation steps are as follows: Take 1g of fresh soil sample and 9ml of sterilized 0.9% normal saline by shaking and mixing, take the supernatant and inoculate it into 100ml of sterilized strain selection liquid medium, culture with shaking at 30℃ for 7 days to enrich the bacteria Kind. Then dilute the coated bacterial solution with 0.9% saline, and add 10 -2 ,10 -3 For the two gradients, 200 μL was applied to the solid plate of the screening medium, and the colony morphology of the growing strain was observed after 3 days of static culture at 30°C. The colony is large and thick, with neat edges, and a single bacteria that seems to have exopolysaccharide production. After Gram staining, 16S rDNA gene sequence (see sequence table) analysis and BIOLOG microbial classification and identification system, the strain is determined...

Embodiment 2

[0039] Example 2: Co-production of biopolysaccharides and poly-β-hydroxybutyric acid in shake flasks

[0040] (1) Inoculate a single colony of Sphingomonas T-3 into 5ml of TPG liquid medium, and culture with shaking at 30°C for 24 hours;

[0041] (2) Inoculate the culture solution prepared in step (1) into 100 ml seed culture medium at an inoculum of 1%, and culture with shaking at 30°C for 24 hours;

[0042] (3) Inoculate the seed liquid prepared in step (2) with a 10% inoculum amount into a glass shaker flask containing 55 ml of fermentation medium, at 30°C, a constant pH of 7.2, and cultivate for 72 hours;

[0043] (4) Dilute the fermentation broth of step (3) with distilled water at least 5 times the volume, heat it at 90-105°C for 15 minutes, and separate the fermentation broth and bacterial cell precipitation by membrane;

[0044] (5) Adjust the pH of the supernatant after membrane separation in step (4) to 3.0 to obtain a jelly-like precipitate, adjust the precipitate to neutral ...

Embodiment 3

[0057] Example 3: Using a 5L fermentor to jointly produce biopolysaccharides and poly-β-hydroxybutyric acid

[0058] (1) Inoculate a single colony of Sphingomonas T-3 into 5ml of TPG liquid medium, and culture with shaking at 30°C for 24 hours;

[0059] (2) Inoculate the culture solution prepared in step (1) into 300 ml seed culture medium at an inoculum amount of 1%, and culture with shaking at 30°C for 24 hours;

[0060] (3) Inoculate the seed liquid prepared in step (2) with an inoculum amount of 10% into a 5L fermentor containing 3.5L of fermentation medium at 30°C and a constant pH of 7.2 for 72 hours;

[0061] (4) Dilute the fermentation broth of step (3) with distilled water at least 5 times the volume, heat it at 90-105°C for 15 minutes, and separate the fermentation broth and bacterial cell precipitation by membrane;

[0062] (5) Adjust the pH of the supernatant after membrane separation in step (4) to 3.0 to obtain a jelly-like precipitate, adjust the precipitate to neutral pH...

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Abstract

The invention relates to a Sphingomonas sp. T-3 and a method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation. The collection number of the Sphingomonas sp. strain T-3 is CGMCC No.10150. The method comprises the following steps: carrying out activating culture on the strain T-3, inoculating into a seed culture medium, inoculating a seed solution into the fermentation culture medium, and culturing; and after the fermentation finishes, diluting with distilled water, heating, carrying out membrane separation to obtain a supernate and a thallus precipitate; regulating the pH value of the supernate to 3.0 or so with hydrochloric acid, regulating the pH value of the obtained precipitate to neutrality with NaOH, and drying to obtain the biological polysaccharide; and drying the thallus precipitate, extracting with chloroform, and drying the supernate to obtain the poly-beta-hydroxy-butanoic acid. In the strain T-3 fermentation process, abundant poly-beta-hydroxy-butanoic acid is accumulated in the cells, and abundant biological polysaccharide is secreted into the culture medium; and the two obtained products can be widely used in the fields of industry, food, medical treatment and the like.

Description

Technical field [0001] The invention belongs to the field of biotechnology and biomaterials, and specifically relates to a method for simultaneously synthesizing biopolysaccharides and poly-β-hydroxybutyric acid using a method of microbial fermentation. Background technique [0002] Sphingomonas sp is a new genus proposed based on the 16s rRNA sequence, respiratory quinone species and cellular polar lipid pattern. The important feature that distinguishes it from other Gram-negative bacteria is that its cell membrane contains glycosphingolipids instead of lipopolysaccharides. Sphingomonas is widely distributed in water, soil and air. At present, the research on Sphingomonas is mainly focused on its ability to degrade refractory organic pollutants. For example, diphenylfuran can be used by Sphingomonas sp.RW1 as the only carbon source material; it has synthetic β carrot Capsule polysaccharides; have the ability to synthesize a class of acidic capsular polysaccharides. These capsu...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/04C12P7/62C12R1/01
CPCC12P7/625C12P19/04C12N1/205C12R2001/01
Inventor 马挺吴萌萌李国强张文文陈思彬周洁芳
Owner NANKAI UNIV
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