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2 '-fucosyllactose high-yield strain and preparation method and application thereof

A technology of fucosyllactose and high-yielding strains, applied in the directions of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as inability to achieve HMO titer, etc.

Pending Publication Date: 2020-07-31
江苏华燕集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, HMO titers greater than 20 g / L in the fermentation broth are not or rarely achieved even with the most efficient methods currently available based on bacterial fermentation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0029] Medium:

[0030] The seed medium is: sucrose 5.0 g / L, nitrogen source and trace elements are composed of: 1.0 g / L yeast extract, 2.0 g / LNH 4 Cl, 10.0 g / L Na 2 HPO 4 ·7H 2 O, 3.0 g / L KH 2 PO 4 , 0.5 g / L NaCl, 0.25 g / L MgSO 4 ·7H 2 O, 15.0 mg / L CaCl 2 2H 2 O and 10 mg / L vitamin B1.

[0031] The fermentation medium is: sucrose 22.0 g / L, 13.5 g / L KH 2 PO 4 , 4.0 g / L (NH 4 ) 2 h 2 PO 4 , 1.7 g / L citric acid, 1.4 g / L MgSO 4 ·7H 2 O, 10 mg / L trace element solution (10 g / L ferric citrate, 2.25 g / LZnSO 4 ·7H 2 O, 10 g / L CuSO 4 ·5H 2 O, 0.35 g / L MnSO 4 ·H 2 O, 0.23 g / L Na 2 B 4 o 7 10H 2 O, 0.11g / L (NH 4 ) 6 Mo 7 o 24 , 2.0 g / L CaCl 2 2H 2 O)

[0032] Analytical method

[0033] Bacterial concentration detection was performed using a spectrophotometer (Biomate 5, Thermo, N.Y., USA). Intracellular FucT2 enzyme protein was detected using 12% SDS-PAGE. After adding 0.1 mM IPTG for 3 h, the cells were collected and the concentration was adjusted to ...

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PUM

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Abstract

The invention discloses a 2 '-fucosyllactose high-yield strain as well as a preparation method and application thereof. The preservation number of the strain is CGMCC No.19557. Escherichia coli E.coliBL21 (DE3) is used as a production host, and phosphomannose mutase (manB), mannose-1-phosphate phosphate guanylyltransferase (manC), GDP-mannose-4, 6-dehydratase (gmd), GDP-L-fucose synthase (flc) and alpha-1,2- fucosyltransferase (futC) are overexpressed so as to construct a synthesis path of the 2'-FL; substrate supply is improved by overexpressing sucrose transporter and lactose transporter onthe basis, the production capacity of 2 '-FL is further improved, efficient expression of 2'-FL in escherichia coli is realized, the 2 '-FL high-yield strain is improved.

Description

technical field [0001] The invention relates to a 2'-fucosyllactose high-yield bacterial strain and its preparation method and application. Background technique [0002] Human milk is a complex mixture of carbohydrates, fats, proteins, vitamins, minerals and trace elements. The main constituents are carbohydrates, which can be further divided into lactose and more complex oligosaccharides (human milk oligosaccharides, HMOs). While lactose can be used by the body as an energy source, complex oligosaccharides cannot be metabolized by infants. Complex oligosaccharides are composed of more than 200 different oligosaccharides, which can constitute up to 20% of the total carbohydrate content. The presence and concentration of these complex oligosaccharides are unique to humans and have not been found in the emulsions of other mammals. [0003] About 200 HMOs with various structures have been identified so far, and many beneficial properties of them have been reported. HMOs can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/70C12N15/90C12N15/66C12N15/61C12N15/60C12N15/54C12N15/53C12N15/31C12P19/00C12R1/19
CPCC12N15/52C12N9/90C12N9/1241C12N9/88C12N9/0006C12N9/1051C07K14/32C07K14/245C12N15/70C12N15/902C12N15/66C12P19/00C12Y504/02008C12Y207/07013C12Y402/01047C12Y101/01271
Inventor 帅玉英荣德明
Owner 江苏华燕集团有限公司
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