CN105002237A - Method for synthesizing L-ascorbic acid-2-glucoside through enzyme fermentation process - Google Patents

Abstract

The invention relates to a method for synthesizing L-ascorbic acid-2-glucoside through the enzyme fermentation process. The method is characterized by comprising the steps of A, adding the purified water in a reaction vessel, then adding vitamin C and starting stirring; B, controlling the temperature in the reaction vessel at 37 DEG C, and adjusting the pH value in the reaction vessel to be 5.50+/-0.10 by the NaOH solution of 30% in mass percentage; C, adding beta-cyclodextrin into the reaction vessel, then adding cyclodextrin glucantransferase and replenishing an appropriate amount of purified water at the same time; D, controlling the temperature in the reaction vessel at 37+/-2 DEG C and reacting in the dark condition for 10-12 hours; E, adding glucoamylase, controlling the temperature in the reaction vessel at 37+/-2 DEG C and reacting in the dark condition for 10-14 hours; F, filtering; G, concentrating; H, crystallizing and drying to obtain the finished product of L-ascorbic acid-2-glucoside. The obtained L-ascorbic acid-2-glucoside is stable and uniform in crystal form, fewer in impurities, high in product purity, environment-friendly and pollution-free.

Description

A kind of method of enzymic fermentation synthesis L-AA-2-glucoside

Technical field

The present invention relates to enzymic fermentation technical field, particularly the method for enzymic fermentation synthesis L-AA-2-glucoside.

Background technology

Vitamins C (xitix) is exactly a kind of conventional whitening raw material.It can suppress the increase of pigment, makes freckle thin out, has whitening effect, can reduce pore, adjustment skin texture.Vitamins C can the generation of check melanin, and melanochrome is once produce, and vitamins C also can make it thin out; please take in vitamins C more; from preventing nti-freckle, protect skin, vitamins C, except having whitening function, increases the effect of collagen protein, adjustment skin texture in addition.

Vitamins C (xitix) can be used for making skin brighter, more healthy, and younger effect widely people is known.Scientific research shows that effect that vitamins C highlights skin whitening comes from its check melanin and formed and reduce melanic ability.But vitamins C is applied to health & beauty formula and faces lot of challenges, be heated, oxidation, with the reaction of other conventional compositions in metal ion and makeup, can make vitamins C lose biological activity easily.This can cause the variable color in cosmetic formulations of its composition, and the more healthy younger effect of skin that makes that it is brought is had a greatly reduced quality.For addressing this problem, there is a kind of special stable vitamins C in the market, being called L-AA 2-glucoside.This vitamin C derivatives, trade(brand)name, referred to as AA-2G, possesses remarkable formulation stability, can inherently suppress variable color and degraded, remain all biological activity simultaneously, highlight whitening to reach, resisting ultraviolet irradiation and antidotal effect.

Current L-AA-2-glucoside mainly adopts the method for chemosynthesis, and environmental hazard is large, and not environmentally, and productive rate is lower.

Summary of the invention

The technical problem to be solved in the present invention is, provide a kind of enzymic fermentation to synthesize the method for L-AA-2-glucoside, the method has environmental protection, the advantage that productive rate is high.

For solving the problems of the technologies described above, the present invention by the following technical solutions: the method for a kind of enzymic fermentation synthesis L-AA-2-glucoside, is characterized in that adopting following steps:

A, in reactor, add pure water, then add vitamins C, open and stir;

In B, control reactor, temperature is at 37 DEG C, regulates pH to 5.50 ± 0.10 by the NaOH solution of mass percent 30%;

C, in described reactor, add beta-cyclodextrin, and then add Maltose 4-glucosyltransferase, add appropriate amount of purified water simultaneously;

D, control reactor temperature are at 37 ± 2 DEG C, and lucifuge reacts 10-12h;

E, add saccharifying enzyme, control control temperature at 37 ± 2 DEG C, lucifuge reaction 10-14h;

F, in reactor, add pure water rinse, after flushing mixed solution; Mixed solution is carried out centrifugation, then filters, obtain filtrate;

G, transfer in reactor by gained filtrate, reduce pressure when temperature≤50 DEG C, concentrated during vacuum tightness≤-0.1MPa, being concentrated into AA 2G content is more than 60%;

H, control still temperature are to 38-45 DEG C, and add crystal seed, stir, then leave standstill slow crystallization, slow cooling 5h, temperature is down to 5-10 DEG C, opens and stirs, and then 5-10 DEG C is incubated crystallization;

J, crystallization are complete, and blowing is centrifugal, centrifugally to spray drip washing with frozen water after dripless flows out, and continue centrifugal, centrifugally after dripless flows out, continue centrifugal 30min, and discharging obtains AA 2G wet product; In temperature≤50 DEG C, dry under vacuum tightness <-0.1Mpa environment, after drying, obtain AA 2G finished product.

The present invention adopts enzyme process to prepare L-AA-2-glucoside, and the AA 2G crystal habit stable and uniform of precipitation, impurity is few, and product purity is high, environment friendly and pollution-free.

Embodiment

Embodiment 1:

A method for enzymic fermentation synthesis L-AA-2-glucoside, is characterized in that adopting following steps:

A, in reactor, add pure water, then add vitamins C, open and stir;

In B, control reactor, temperature is at 37 DEG C, regulates pH to 5.50 ± 0.10 by the NaOH solution of mass percent 30%;

C, in described reactor, add beta-cyclodextrin, and then add Maltose 4-glucosyltransferase, add appropriate amount of purified water simultaneously;

D, control reactor temperature are at 35 DEG C, and lucifuge reacts 10h;

E, add saccharifying enzyme, control control temperature at 35 DEG C, lucifuge reaction 10h;

F, in reactor, add pure water rinse, after flushing mixed solution; Mixed solution is carried out centrifugation, then filters, obtain filtrate;

G, transfer in reactor by gained filtrate, reduce pressure when temperature≤50 DEG C, concentrated during vacuum tightness≤-0.1MPa, being concentrated into AA 2G content is more than 60%;

H, control still temperature to 38 DEG C, add crystal seed, stir, then leave standstill slow crystallization, slow cooling 5h, temperature is down to 5 DEG C, opens and stirs, and then 5 DEG C are incubated crystallizatioies;

J, crystallization are complete, and blowing is centrifugal, centrifugally to spray drip washing with frozen water after dripless flows out, and continue centrifugal, centrifugally after dripless flows out, continue centrifugal 30min, and discharging obtains AA 2G wet product; In temperature≤50 DEG C, dry under vacuum tightness <-0.1Mpa environment, after drying, obtain AA 2G finished product.

Embodiment 2:

A method for enzymic fermentation synthesis L-AA-2-glucoside, is characterized in that adopting following steps:

A, in reactor, add pure water, then add vitamins C, open and stir;

In B, control reactor, temperature is at 37 DEG C, regulates pH to 5.50 ± 0.10 by the NaOH solution of mass percent 30%;

C, in described reactor, add beta-cyclodextrin, and then add Maltose 4-glucosyltransferase, add appropriate amount of purified water simultaneously;

D, control reactor temperature are at 39 DEG C, and lucifuge reacts 12h;

E, add saccharifying enzyme, control control temperature at 39 DEG C, lucifuge reaction 14h;

F, in reactor, add pure water rinse, after flushing mixed solution; Mixed solution is carried out centrifugation, then filters, obtain filtrate;

G, transfer in reactor by gained filtrate, reduce pressure when temperature≤50 DEG C, concentrated during vacuum tightness≤-0.1MPa, being concentrated into AA 2G content is more than 60%;

H, control still temperature to 45 DEG C, add crystal seed, stir, then leave standstill slow crystallization, slow cooling 5h, temperature is down to 10 DEG C, opens and stirs, and then 10 DEG C are incubated crystallizatioies;

J, crystallization are complete, and blowing is centrifugal, centrifugally to spray drip washing with frozen water after dripless flows out, and continue centrifugal, centrifugally after dripless flows out, continue centrifugal 30min, and discharging obtains AA 2G wet product; In temperature≤50 DEG C, dry under vacuum tightness <-0.1Mpa environment, after drying, obtain AA 2G finished product.

Embodiment 3:

A method for enzymic fermentation synthesis L-AA-2-glucoside, is characterized in that adopting following steps:

A, in reactor, add pure water, then add vitamins C, open and stir;

In B, control reactor, temperature is at 37 DEG C, regulates pH to 5.50 ± 0.10 by the NaOH solution of mass percent 30%;

C, in described reactor, add beta-cyclodextrin, and then add Maltose 4-glucosyltransferase, add appropriate amount of purified water simultaneously;

D, control reactor temperature are at 37 DEG C, and lucifuge reacts 11h;

E, add saccharifying enzyme, control control temperature at 37 DEG C, lucifuge reaction 11h;

F, in reactor, add pure water rinse, after flushing mixed solution; Mixed solution is carried out centrifugation, then filters, obtain filtrate;

G, transfer in reactor by gained filtrate, reduce pressure when temperature≤50 DEG C, concentrated during vacuum tightness≤-0.1MPa, being concentrated into AA 2G content is more than 60%;

H, control still temperature to 40 DEG C, add crystal seed, stir, then leave standstill slow crystallization, slow cooling 5h, temperature is down to 8 DEG C, opens and stirs, and then 8 DEG C are incubated crystallizatioies;

J, crystallization are complete, and blowing is centrifugal, centrifugally to spray drip washing with frozen water after dripless flows out, and continue centrifugal, centrifugally after dripless flows out, continue centrifugal 30min, and discharging obtains AA 2G wet product; In temperature≤50 DEG C, dry under vacuum tightness <-0.1Mpa environment, after drying, obtain AA 2G finished product.

Claims (1)

1. a method for enzymic fermentation synthesis L-AA-2-glucoside, is characterized in that adopting following steps:

A, in reactor, add pure water, then add vitamins C, open and stir;

In B, control reactor, temperature is at 37 DEG C, regulates pH to 5.50 ± 0.10 by the NaOH solution of mass percent 30%;

C, in described reactor, add beta-cyclodextrin, and then add Maltose 4-glucosyltransferase, add appropriate amount of purified water simultaneously;

D, control reactor temperature are at 37 ± 2 DEG C, and lucifuge reacts 10-12h;

E, add saccharifying enzyme, control control temperature at 37 ± 2 DEG C, lucifuge reaction 10-14h;

F, in reactor, add pure water rinse, after flushing mixed solution; Mixed solution is carried out centrifugation, then filters, obtain filtrate;

G, transfer in reactor by gained filtrate, reduce pressure when temperature≤50 DEG C, concentrated during vacuum tightness≤-0.1MPa, being concentrated into AA 2G content is more than 60%;

H, control still temperature are to 38-45 DEG C, and add crystal seed, stir, then leave standstill slow crystallization, slow cooling 5h, temperature is down to 5-10 DEG C, opens and stirs, and then 5-10 DEG C is incubated crystallization;

J, crystallization are complete, and blowing is centrifugal, centrifugally to spray drip washing with frozen water after dripless flows out, and continue centrifugal, centrifugally after dripless flows out, continue centrifugal 30min, and discharging obtains AA 2G wet product; In temperature≤50 DEG C, dry under vacuum tightness <-0.1Mpa environment, after drying, obtain AA 2G finished product.