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Method for separating and purifying fructooligosaccharides

A technology for separation and purification of fructooligosaccharides, applied in the directions of oligosaccharides, chemical instruments and methods, microorganism-based methods, etc., can solve the problem of high cost of fructosyltransferase, and achieve the effect of high product content

Active Publication Date: 2014-06-04
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problem of using fructosyltransferase to produce fructooligosaccharides in industry is the high cost of fructosyltransferase in the enzymatic hydrolysis of sucrose

Method used

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  • Method for separating and purifying fructooligosaccharides
  • Method for separating and purifying fructooligosaccharides
  • Method for separating and purifying fructooligosaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Preparation of fructooligosaccharide fermentation broth

[0059] Phaffia strain JMU-MVP14 with high astaxanthin production was selected and bred. The volumetric yield of astaxanthin fermented by this strain reached 150 mg / L, and the cell yield of astaxanthin reached 6 mg / g. The fructo-oligosaccharide fermented liquid obtained by the fermentation of sucrose by Phaffia yeast JMU-MVP14, the specific preparation method is as follows:

[0060] Inoculate 1 ml of activated Phaffia yeast JMU-MVP14 into shaker flask culture medium, and culture at 22°C, 190 r / min for 72 h to obtain seed liquid; In a 7 L fermentation tank with L fermentation medium, the temperature was controlled at 22°C, the ventilation rate was 3 L / min, and the dissolved oxygen was controlled at 30-40% by adjusting the stirring speed; 25% ammonia water was automatically fed to control the pH 6.0; every 8 Take a sample once to measure the astaxanthin content. After the astaxanthin synthesis reaches ...

Embodiment 2

[0064] The filtrate containing fructooligosaccharides was obtained according to the method in Example 1. Ion exchange resin was used as an adsorption medium to separate fructo-oligosaccharides.

[0065] Put the pretreated ion exchange resin (SP-Sepharose resin) into the chromatographic column with a column volume of 40 ml. After the resin settles, keep the column temperature at 25°C and equilibrate with ultrapure water for 5 column volumes. According to the method in Example 1, the filtrate containing fructooligosaccharides was loaded on the chromatographic column with a flow rate of 1 ml / min, and the loading volume was 40 ml, and then washed with ultrapure water, and the flow rate was 1 ml / min. min, collect one tube per 5 ml, detect with HPLC, collect the collection solution containing fructooligosaccharides, load the sample on the Q-Sepharose column chromatography with ultrapure water to balance 5 column volumes, wash with ultrapure water, The flow rate is 1 ml / min, one tub...

Embodiment 3

[0067] The filtrate containing fructooligosaccharides was obtained according to the method in Example 1.

[0068] Weigh an equal mass of pretreated diatomaceous earth and activated carbon and mix them into a chromatographic column with a volume of 40 ml. After it settles, keep the column temperature at 25°C and equilibrate 5 column volumes with ultrapure water Afterwards, the filtrate containing fructooligosaccharides obtained by the method in Example 1 is loaded on the chromatographic column with a flow rate of 1 ml / min, and the loading volume is 40 ml, and then washed with ultrapure water, and the conductivity is After the value reaches a stable value, use 50 ml of ethanol with a volume fraction of 5%, 10%, 15% and 20% to elute step by step, the flow rate is 1 ml / min, collect one tube for every 5 ml, and detect the oligomerization For fructose content, collect the collected liquid containing fructooligosaccharides, concentrate it to the volume before loading the sample by ro...

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Abstract

The invention discloses a method for separating and purifying fructooligosaccharides, wherein separating and purifying steps include: fermenting cane sugar through Phaffia rhodozyma JMU-MVP14 to obtain a fermentation fluid containing the fructooligosaccharides and astaxanthin; centrifugally separating the fermentation fluid through a centrifuge, extracting the lower-layer astaxanthin, heating the supernatant fluid through a boiling water bath for enzyme inactivation and then purifying the supernatant fluid through a microfiltration membrane, wherein the obtained filtrate flows through a chromatographic column filled with an absorption medium to be absorbed; next, eluting the absorption medium using ethanol solution; and evaporating and concentrating the eluate to obtain the fructooligosaccharides. The method is capable of simultaneously separating and purifying the fructooligosaccharides and the astaxanthin in the same batch of fermentation process; the separation operation is low in cost and simple; the obtained fructooligosaccharide product is high in content and meets the requirements of food sanitation.

Description

technical field [0001] The invention belongs to the field of fructo-oligosaccharide synthesis by fermentation method, and in particular relates to a separation and purification method of fructo-oligosaccharide synthesized by fermentation of Phaffia yeast JMU-MVP14. Background technique [0002] Fructose-oligosaccharides are formed by linking the fructose residues of sucrose molecules with one or more fructosyl groups through β-(1,2)-glucosidic bonds. FOS is easily soluble in water, its sweetness is 30%-60% of that of sucrose, its moisture retention is higher than that of sucrose, equivalent to sorbitol, its viscosity is higher than that of sucrose at the same concentration, and it has a strong pH value between 4.0-7.0 thermal stability. [0003] As people's health awareness becomes stronger and stronger, the market demand for functional food is increasing. In 2005, the market value of functional food in the United States was about 73.5 billion US dollars, and the demand in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P23/00C12P19/00C07H3/06C07H1/06C07C403/24C12R1/645
Inventor 肖安风李利君倪辉蔡慧农杨远帆黄高凌杜希萍
Owner JIMEI UNIV
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