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Low-impurity high-yielding gellan gum-producing bacteria and its application

A technology for gellan gum and bacteria production, which is applied in the direction of bacteria, microorganisms, microorganisms, etc., to achieve the effect of reducing purification cost, increasing yield and reducing production cost

Active Publication Date: 2020-12-01
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are currently many methods of genetic engineering to increase production and reduce metabolic by-products, gellan gum, as a food additive, is produced using genetically engineered strains, especially when food safety issues are extremely prominent, which is likely to cause consumers' concerns

Method used

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  • Low-impurity high-yielding gellan gum-producing bacteria and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Screening of gellan gum-producing bacteria without nodocaxanthin (EMS mutagenesis and high-efficiency screening):

[0028] 1), the first round of mutagenesis treatment:

[0029] The slant of the sphingomonas elatiens ATCC 31461YM medium preservation was transferred to the LB liquid medium, cultivated at 30°C for 16h, and then added EMS (ethyl methanesulfonate) according to the volume ratio of 1% of the LB liquid medium to avoid Leave at room temperature for 1 hour, then transfer to fresh YM liquid medium, and incubate at 30°C for 6 hours.

[0030] The composition of YM liquid medium: peptone 5g / L, malt extract powder 3g / L, glucose 10g / L, yeast powder 3g / L, solvent is water, pH7.0.

[0031] The composition of YM solid medium: peptone 5g / L, malt extract powder 3g / L, glucose 10g / L, yeast powder 3g / L, 15g / L agar, solvent is water, pH7.0.

[0032] 2) The samples obtained from the first round of mutagenesis treatment were spread on the YM plate (YM solid medium) ...

Embodiment 2

[0034] Example 2, Screening of gellan gum-producing bacteria without nodocaxanthin and PHB:

[0035] 1), the second round of mutagenesis treatment:

[0036] The strain obtained in Example 1 with the highest gellan gum polysaccharide yield and no nodocaxanthin production was used as the starting strain for the second round of mutagenesis, transferred to YM liquid medium, cultured at 30°C for 16h, and then followed the YM liquid medium Add EMS at a volume ratio of 1%, place at room temperature for 1 h in the dark, then transfer to fresh YM medium, and incubate at 30°C for 6 h.

[0037] 2) The samples obtained from the second round of mutagenesis treatment were serially diluted and spread on YM solid medium added with 5 μg / mL Nile blue Nile blue fluorescent agent, and cultured at 30° C. for 72 hours. Among them, Nile blue can be combined with PHB to make the PHB-containing strains produce fluorescence under ultraviolet light, while the non-fluorescence or extremely weak fluoresc...

Embodiment 3

[0039] Example 3, Screening of producing bacteria that do not contain nodocaxanthin and PHB and high-yield gellan gum:

[0040] 1), the third round of mutagenesis treatment:

[0041]The strain obtained in Example 2 with the highest polysaccharide yield and no nodocaxanthin and PHB was the starting strain for the third round of mutagenesis, transferred to YM liquid medium, cultivated at 30°C for 16h, and then followed the method of 1% YM liquid medium. Add EMS to the volume ratio, place at room temperature in the dark for 1 hour, transfer to fresh YM medium, and incubate at 30°C for 6 hours;

[0042] 2) The samples obtained from the third round of mutagenesis treatment were serially diluted and spread on a YM high-yield screening plate supplemented with 100 μg / mL ampicillin, and cultured at 30° C. for 5 days.

[0043] Recipe for YM high yield screening plate supplemented with 100 μg / mL ampicillin:

[0044] Peptone 5g / L, malt extract powder 3g / L, glucose 10g / L, yeast powder 3g...

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Abstract

The invention discloses a low-impurity and high-yield gellan gum-producing bacterium, which is Sphingomonas elodea LG010, and its preservation number is: CCTCC NO: 2016143. The present invention also provides the use of the low-impurity, high-yield gellan gum-producing bacteria: producing gellan gum that does not contain nodocaxanthin and poly-β-hydroxybutyric acid. The invention removes the metabolic by-products that compete with gellan gum for substrates and affect purification from the source, effectively improves fermentation efficiency, simplifies purification steps, and reduces production costs.

Description

technical field [0001] The invention relates to a low-impurity high-yield gellan gum-producing bacterium and its application. Background technique [0002] As a fermentation industrial product, microbial polysaccharides not only have excellent performance but also have many special functions that other similar polysaccharides lack. They are more widely used than animal and plant polysaccharides, and their production is not affected by factors such as geographical environment, climate, and natural disasters, and the production cycle is short. , stable output and quality, high cost performance. To a large extent, it can meet people's demand for natural pollution-free food. [0003] Gellan gum is a new type of microbial polysaccharide, which was discovered in 1978. It is a linear anionic heteropolysaccharide produced by Sphingomonas elacha under aerobic conditions. It has been approved as a food additive to be used in various foods in Japan, the United States, the European Un...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/04C12R1/01
CPCC12P19/04C12N1/205C12R2001/01
Inventor 李昂李欧胡婷婷骆航琦忻佳琪栗宏伟林伊诺黄敬瑜郑文倩
Owner ZHEJIANG SCI-TECH UNIV
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