Green preparation method of high-purity galactooligosaccharide

A high-purity galacto-oligosaccharide technology, which is applied in the field of green preparation of high-purity galacto-oligosaccharides, can solve the problems of unfavorable large-scale production of high-purity GOS, high equipment requirements, and complicated operations, so as to be beneficial to industrial applications and The effects of promotion, fast growth and metabolic rate, and simple operation

Inactive Publication Date: 2017-12-29
TIANJIN UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods require high equipment, complicated operation and high cost, and the purity of the product usually obtained is not more than 65%, which is not conducive to large-scale production of high-purity GOS

Method used

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preparation example Construction

[0038] The green preparation method of high-purity galacto-oligosaccharide of the present invention ( figure 1 ), including the following steps:

[0039] 1) Fermentation preparation of whole yeast cells containing β-galactosidase intracellular enzyme

[0040] Inoculate yeast single clones into the seed liquid medium to prepare the yeast seed liquid, and then add 0.1-10% (v / v) yeast seed liquid to the lactose-rich biomass medium at a temperature of 16-37°C, stirring at a rotating speed It is prepared by fermentation under the conditions of 50-500 rpm and pH value of 4-10, and the fermentation time is 8 to 48 hours, and a fermentation broth containing whole yeast cells with a concentration of 1-20 g / L can be obtained.

[0041] 2) Use ethanol solution to permeabilize whole yeast cells

[0042] The above-mentioned fermentation broth is centrifuged to obtain yeast whole cell sediment and fermentation supernatant, which is used for permeabilization treatment and reaction purification of cru...

Embodiment 1

[0048] Fermentation Preparation of Pichia Pastoris Whole Cells Containing β-Galactosidase Intracellular Enzyme

[0049] The preserved Pichia pastoris strain was inoculated onto a flat solid LB medium and cultured at 30°C for 72 hours to obtain activated single colonies. The activated single colony was inoculated into liquid LB medium, and it was shaken and expanded for 20 hours at 30° C. and 160 rpm to obtain seed culture solution. In a 500mL Erlenmeyer flask, connect 0.1mL seed culture broth to 100mL fermentation medium, culture for 48h at the initial pH value of 4.0, rotating speed 50rpm, temperature 16℃, to obtain fermentation broth, and centrifuge to obtain whole Pichia yeast cells Precipitation and fermentation supernatant, Pichia pastoris whole cell precipitation is used for permeabilization and reaction purification of crude galactooligosaccharide (GOS), and the fermentation supernatant is removed. The formula of the fermentation medium is as follows: whey powder 100g / L, ...

Embodiment 2

[0057] Fermentation preparation of whole cells of Saccharomyces cerevisiae containing β-galactosidase intracellular enzyme

[0058] The preserved Saccharomyces cerevisiae strains were inoculated on a flat solid LB medium and cultured at 30°C for 72 hours to obtain activated single colonies. The activated single colony was inoculated into liquid LB medium, and it was shaken and expanded for 20 hours at 30° C. and 160 rpm to obtain seed culture solution. In a 500mL Erlenmeyer flask, insert 10mL seed culture solution into 100mL fermentation medium, culture for 8h at the initial pH value of 10, rotating speed of 500rpm, and temperature of 37℃ to obtain fermentation broth, and centrifuge to obtain the whole yeast Cell precipitation and fermentation supernatant, Saccharomyces cerevisiae whole cell precipitation is used for permeabilization and reaction purification of crude GOS, and fermentation supernatant is removed. The formula of the fermentation medium is as follows: whey 100mL / L...

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Abstract

The invention discloses a green preparation method of high-purity galactooligosaccharide. The green preparation method comprises the following steps of fermenting preparation of yeast whole cell containing beta-galactosidase endoenzyme, permeabilizing treatment of yeast whole cell by an ethanol solution, preparation of galactooligosaccharide by permeabilized yeast cell catalyzing lactose (purpose of yeast I), and obtaining of high-purity galactooligosaccharide (purpose of yeast II) by using yeast whole cell reaction to purify a crude product of galactooligosaccharide. The green preparation method has the advantages that the permeable yeast cell and the yeast whole cell can be repeatedly utilized, so that the effect of one yeast, two purposes is realized in the green cycling preparation of the high-purity galactooligosaccharide; the purity of the obtained high-purity galactooligosaccharide can reach 99% to the highest degree, the technology is simple, no harmful matter is added, the required equipment cost is low, and the high-purity galactooligosaccharide is suitable for being widely and industrially popularized.

Description

Technical field [0001] The invention belongs to the technical field of preparation of functional oligosaccharides, and relates to a green preparation method of high-purity galacto-oligosaccharides, including the fermentation preparation of whole yeast cells containing β-galactosidase intracellular enzyme, and the use of ethanol solution Permeabilization treatment of yeast whole cells, including the use of permeabilized yeast cells to catalyze lactose to prepare galacto-oligosaccharides, and also including the use of yeast whole-cell reaction to purify crude galacto-oligosaccharides to obtain high-purity galacto-oligosaccharides, and the method is transparent Both the sexualized yeast cells and whole yeast cells can be recycled repeatedly, realizing the green recycling preparation of high-purity galactooligosaccharides of "one type of yeast, two uses". Background technique [0002] Galacto-oligosaccharides (GOS) is an important prebiotic factor, which can effectively promote the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/00C12N1/16C12N1/18C12N1/00C07H1/06C07H3/06C12R1/84C12R1/865C12R1/645
CPCC07H1/06C07H3/06C12N1/005C12N1/16C12N1/18C12P19/00
Inventor 齐崴王梦凡尤生萍苏荣欣
Owner TIANJIN UNIV
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