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Batch fermentation method of 2'-fucosyllactose

A batch fermentation technology of fucosyllactose, which is applied in the field of microbial fermentation, can solve the problems of instability, yield not as good as that of E. coli system, complex system, etc., and achieve easy expansion and high 2'-fucosyllactose production , the effect of simple components

Inactive Publication Date: 2021-04-16
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, 2’-FL is also produced by constructing Bacillus subtilis and Corynebacterium glutamicum, but because the system is complex and unstable, the yield is far inferior to that of the E. coli system

Method used

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  • Batch fermentation method of 2'-fucosyllactose
  • Batch fermentation method of 2'-fucosyllactose
  • Batch fermentation method of 2'-fucosyllactose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Use a displacement fermenter with a volume of 5L to carry out the fermentation operation, and the liquid filling capacity is 3L.

[0043]1) Preparation of primary seed fermentation broth: Escherichia coli E.coli-XYY1, a glycerol strain frozen in a -80 refrigerator, was inoculated in 50 mL of LB medium at a 2% inoculum size, and cultured on a shaker at 37° C. and 220 rpm for 8 hours.

[0044] 2) Preparation of secondary seed fermentation broth: 30 mL of primary seed fermentation broth was inoculated into 300 mL of LB medium, and cultured on a shaking table at 37° C. and 220 rpm for 4 hours.

[0045] 3) Fermentation tank medium configuration: add Na to the 5L tank 2 HPO 4 12H 2 O 51.3g, NaCl 1.5g, KH 2 PO 4 9g, NH 4 Cl 12g, Triton 0.3mL, Tween-800.3mL, dilute to 2L with deionized water, sterilize in a high-temperature sterilization pot at 121°C for 15min, and then cool to 37°C. Also configure MgSO 4 72mg in 10mL deionized water, CaCl 2 13.5mg in 10mL of deionize...

Embodiment 2

[0049] A 30L capacity in-situ fermenter was used for the fermentation operation, and the liquid loading capacity was 20L.

[0050] 1) Preparation of first-grade seed fermentation broth: Escherichia coli E.coli-XYY1, a glycerol strain frozen in a -80 refrigerator, was inoculated in 100 mL of LB medium at a 5% inoculum size, and cultured on a shaking table at 37° C. and 220 rpm for 6.5 hours.

[0051] 2) Preparation of secondary seed fermentation broth: 100 mL of primary seed fermentation broth was inoculated into 2L LB medium, and cultured on a shaker at 37° C. and 220 rpm for 6 hours.

[0052] 3) Fermentation tank medium configuration: add Na to the 30L tank 2 HPO 4 12H 2 O 342g, NaCl 10g, KH 2 PO 4 60g, NH 4 Cl 80g, Triton 2mL, Tween-802mL, dilute to 13L with deionized water, set high temperature steam sterilization at 121°C for 15min, then cool to 37°C. Also configure MgSO 4 480mg in 10mL deionized water, CaCl 2 90mg in 10mL of deionized water, 320g of glucose in 1...

Embodiment 3

[0055] A 30L capacity in-situ fermenter was used for the fermentation operation, and the liquid loading capacity was 20L.

[0056] (1) Preparation of first-grade seed fermentation broth: Escherichia coli E.coli-XYY1, a glycerol strain frozen in a -80 refrigerator, was inoculated in 100 mL of LB medium at a 3% inoculum size, and cultured on a shaking table at 37° C. at 220 rpm for 6 hours.

[0057] (2) Preparation of the secondary seed fermentation liquid: take 100 mL of the primary seed fermentation liquid and inoculate it in 2L LB medium, and culture it on a shaking table at 37° C. at 220 rpm for 4 hours;

[0058] (3) Fermentation tank medium configuration: add Na to the 30L tank 2 HPO 4 12H 2 O 342g, NaCl 10g, KH 2 PO 4 60g, NH 4 Cl 80g, Triton 2mL, Tween-802mL, dilute to 13L with deionized water, set high temperature steam sterilization at 121°C for 15min, then cool to 37°C. Also configure MgSO 4 480mg in 10mL deionized water, CaCl 2 90mg in 10mL of deionized wate...

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Abstract

The invention discloses a batch fermentation method of 2'-fucosyllactose. The method is more suitable for large-scale fermentation on the original basis, is higher in production efficiency and final yield, is easier for industrial enlarged culture, and has higher application prospect and economic value. A fermentation medium used in the method is simple in component, free of toxic and harmful components, economical, cheap and easy to obtain, and an excellent fermentation effect and high 2'- fucosyllactose yield can be obtained.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a fermentation tank batch fermentation method of 2'-fucosyllactose. Background technique [0002] 2'-Fucosyllactose (2'-FL) is the most abundant oligosaccharide in breast milk, with a molecular weight of 488.44Da. It connects the fucose residue and the galactose end of lactose through an α1-2 glycosidic bond, and is a very special natural trisaccharide. As a neutral oligosaccharide, 2'-FL can resist the digestion of human gastrointestinal enzymes, so it can directly reach the colon of the human body, be digested and decomposed by the microorganisms in the human intestinal tract, and promote the proliferation of related beneficial bacteria. Acts as a prebiotic. [0003] In recent years, a large number of studies have shown that oral administration of 2'-FL can regulate the immune function of suckling mice, reduce inflammation, enhance intestinal barrier function...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/00C12N1/38C12R1/19
Inventor 王硕王远一飞
Owner NANKAI UNIV
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