CN105586381A - Preparation method of high-dispersity low-acyl gellan gum - Google Patents

Abstract

The invention discloses a preparation method of high-dispersity low-acyl gellan gum, belonging to the field of low-acyl gellan gum. The method comprises the following steps: inoculating Pseudomonaselodea and Bacillus pumilus into a fermentation culture medium, fermenting, separating the fermentation liquid, taking the supernate, carrying out deproteinization, regulating the pH value, filtering with a cellulose acetate filter membrane, mixing the filtrate with chitosan until flocculation is generated, dissolving, carrying out acyl removal treatment, and finally, carrying out decolorization and spray drying to obtain the high-dispersity low-acyl gellan gum. When the low-acyl gellan gum is dissolved in cold water, no phenomenon of agglomeration can be generated; the low-acyl gellan gum can be uniformly dispersed in cold water even no additive is added; the method has the advantages of simple preparation technique and low cost; and the obtained product has the advantages of high gel strength and high transparency, and has wide application prospects.

Description

A kind of preparation method of high dispersive low-acyl gellan gum

Technical field

The invention discloses a kind of preparation method of high dispersive low-acyl gellan gum, belong to low-acyl gellan gum field.

Background technology

Gellan gum is a kind of line style polyanion microbial polysaccharide, has unique gel characteristic and solution rheoiogy character, and self-discovery rises and is applied in foods and cosmetics. In recent years, along with the development of biomedical subject, natural polymer gellan gum and hydrogel thereof, shown wide application prospect in the field such as drug delivery system and tissue engineering material. Gellan gum is nontoxic, has biocompatibility and biodegradable, the transparent and good stability of the hydrogel that forms, and the mechanical property of gel is close with human body normal tissue under certain condition.

Gellan gum is a kind of anion microbial polysaccharide, there is the good characteristics such as gel formation ability is strong, transparency is high, acidproof heat-proof performance is good in food processing industry, gellan gum not only can be used as a kind of good gelling agent and uses, and can give quality and the mouthfeel of food pleasant; In addition, gellan gum also can be used in conjunction with other foodstuff glues, is widely used in, in the numerous food processing such as millet cake, candy, flavouring, making food can obtain best sense organ, matter structure and stability requirement.

In food industry, the dispersed of hydrophilic colloid is very important, sometimes needs a large amount of stir process to avoid due to colloid swelling insufficient " flake " producing of hydration that cause of uniting, lump in production. In the operation of food industry at present; the dispersiveness that low-acyl gellan gum has been had; the in the situation that of formula and production technology permission, gellan gum and chelating agent, white sugar or other prepare burden dry powder, vegetable oil, propane diols etc. can be mixed in advance, then put in the lump in cold water. This processing can be separated gellan gum fine particle, reaches homodisperse effect in water, but in some special food, can not use other batchings, and this will significantly impact the dispersive property of low-acyl gellan gum.

Summary of the invention

The technical problem that the present invention mainly solves: for being dissolved in separately in cold water and easily uniting at present, must or be dry mixed into other powdery batchings by high-speed stirred and could improve dispersive property, if do not exist in formula in other powdery batchings or operating process and do not allow high-speed stirred, the low dispersiveness of product can badly influence the processing in later stage, and gellan gum productive rate is not high, the problem of step complexity, a kind of preparation method of high dispersive low-acyl gellan gum is provided, the present invention is inoculated into waterweed pseudomonad and bacillus pumilus in fermentation medium and ferments, separation of fermentative broth is got to supernatant de-albumen, after regulating pH value, use cellulose acetate membrane filtration, filtrate and shitosan mixing generation flocculation dissolving are carried out to the processing of deacylated tRNA base, finally decolouring, the dry loose low-acyl gellan gum that can secure satisfactory grades of spraying, the low-acyl gellan gum that the present invention makes is dissolved in cold water, can not produce and unite, also can be dispersed in cold water even if do not add batching, and the gel strength of gellan gum reaches 1200g/cm²Above, transparency reaches more than 89%, is with a wide range of applications.

In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:

(1) count by weight, get the sodium chloride solution of 30～40 parts of glucose, 15～20 parts of peptones, 2～3 parts of Dried Corn Steep Liquor Powder, 18～22 portions of beef extracts, 1.5～2.2 parts of dipotassium hydrogen phosphates, 0.5～1.8 part of potassium dihydrogen phosphate and 21～23 parts of mass fractions 0.5%, after mixing, mixture is placed on to sterilizing under uviol lamp, obtain fermentation medium, the ammoniacal liquor that is 2% with mass fraction regulates fermentation medium pH to 7.0;

(2) fermentation medium of getting after the above-mentioned pH of 1.0～1.5L has regulated is put into 3L fermentation tank, by inoculum concentration 8～10%, mixed bacteria liquid is put into fermentation tank, Temperature Setting is 30～35 DEG C, speed setting is 150～170r/min, ferment 8～10 days, described mixed bacteria liquid is that concentration is 1.5 × 10⁸Waterweed pseudomonad bacterium liquid and the concentration of individual/mL are 0.6 × 10⁸The bacillus pumilus liquid of individual/mL by volume 1:1 mixes;

(3) after above-mentioned fermentation ends, collect zymotic fluid, zymotic fluid is put into centrifuge, centrifugation 5～8min under 3000～5000r/min, collect supernatant, supernatant is put into beaker, in beaker, add supernatant volume 30～35% absolute ethyl alcohols, stir;

(4) after above-mentioned stirring, put into water-bath, with 5 DEG C/min speed, beaker is heated up, in the time that temperature rises to 35～45 DEG C, in beaker, add supernatant quality 2～4% compound proteases, carry out insulated and stirred 1～2h with rotating speed 200r/min, and then continue to be warming up to 85～95 DEG C with 5 DEG C/min speed, after heating up, regulating pH with the potassium hydroxide solution of mass fraction 15% is 10.5～11.0, insulation 50～60min;

(5) after finishing, above-mentioned insulation naturally cools to room temperature, mixture in beaker is filtered with cellulose acetate filter membrane, collect filtrate and put into flask, to the absolute ethyl alcohol that adds 2～3 times of filtrate volumes in flask, add again filtrate quality 2～3% shitosans, be uniformly mixed, leave standstill until filter collection filtrate after producing without flocculate;

(6) filtrate of above-mentioned gained being put into spray dryer sprays dry; collect dry thing and put into beaker; to the water that adds 70 DEG C in beaker; stir until dry thing stops adding water after dissolving completely; again to the 200 order carclazytes that add dry matter amount 6～8% in beaker, stir after 30～40min filtration under diminished pressure while hot, collect filtrate and put into spray dryer and spray and be dried; collect dry thing, can obtain high dispersive low-acyl gellan gum.

The method of application of the present invention: take the gellan gum that 1～2g the present invention makes, be scattered in 250～350mL distilled water, at room temperature be stirred to abundant dissolving, in solution, add respectively 0.4～0.6g calcium chloride, 7～9g jelly powder, 120～140g white granulated sugar, 0.8～1.2g natrium citricum, 0.1～0.2g essence pigment, after stirring, be heated to continuation at 60～80 DEG C and stir 20～30min, after stirring, be cooled to room temperature, after cooling formation gel, can form jelly, at 100～105 DEG C, high-temperature sterilization is processed 10～15min.

The invention has the beneficial effects as follows:

(1), when the low-acyl gellan gum that the present invention makes is dissolved in cold water, can not produces and unite, also can be dispersed in cold water even if do not add batching;

(2) the low-acyl gellan gum gel strength that the present invention generates is high, and transparency is high, and preparation technology is simple, and cost is low, is with a wide range of applications.

Detailed description of the invention

First meter by weight, get the sodium chloride solution of 30～40 parts of glucose, 15～20 parts of peptones, 2～3 parts of Dried Corn Steep Liquor Powder, 18～22 portions of beef extracts, 1.5～2.2 parts of dipotassium hydrogen phosphates, 0.5～1.8 part of potassium dihydrogen phosphate and 21～23 parts of mass fractions 0.5%, after mixing, mixture is placed on to sterilizing under uviol lamp, obtain fermentation medium, the ammoniacal liquor that is 2% with mass fraction regulates fermentation medium pH to 7.0; The fermentation medium of getting after the above-mentioned pH of 1.0～1.5L has regulated is put into 3L fermentation tank, by inoculum concentration 8～10%, mixed bacteria liquid is put into fermentation tank, Temperature Setting is 30～35 DEG C, speed setting is 150～170r/min, ferments 8～10 days, and described mixed bacteria liquid is that concentration is 1.5 × 10⁸Waterweed pseudomonad bacterium liquid and the concentration of individual/mL are 0.6 × 10⁸The bacillus pumilus liquid of individual/mL by volume 1:1 mixes; After fermentation ends, collect zymotic fluid, zymotic fluid is put into centrifuge, centrifugation 5～8min under 3000～5000r/min, collects supernatant, and supernatant is put into beaker, in beaker, adds supernatant volume 30～35% absolute ethyl alcohols, stirs; After stirring, put into water-bath, with 5 DEG C/min speed, beaker is heated up, in the time that temperature rises to 35～45 DEG C, in beaker, add supernatant quality 2～4% compound proteases, carry out insulated and stirred 1～2h with rotating speed 200r/min, and then continue to be warming up to 85～95 DEG C with 5 DEG C/min speed, after heating up, regulating pH with the potassium hydroxide solution of mass fraction 15% is 10.5～11.0, insulation 50～60min; After finishing, insulation naturally cools to room temperature, mixture in beaker is filtered with cellulose acetate filter membrane, collect filtrate and put into flask, to the absolute ethyl alcohol that adds 2～3 times of filtrate volumes in flask, add again filtrate quality 2～3% shitosans, be uniformly mixed, leave standstill until filter collection filtrate after producing without flocculate; The filtrate of gained is put into spray dryer sprays dry; collect dry thing and put into beaker; to the water that adds 70 DEG C in beaker; stir until dry thing stops adding water after dissolving completely; again to the 200 order carclazytes that add dry matter amount 6～8% in beaker, stir after 30～40min filtration under diminished pressure while hot, collect filtrate and put into spray dryer and spray and be dried; collect dry thing, can obtain high dispersive low-acyl gellan gum.

Example 1

First meter by weight, get the sodium chloride solution of 30 parts of glucose, 20 parts of peptones, 3 parts of Dried Corn Steep Liquor Powder, 20 portions of beef extracts, 2.2 parts of dipotassium hydrogen phosphates, 1.8 parts of potassium dihydrogen phosphates and 23 parts of mass fractions 0.5%, after mixing, mixture is placed on to sterilizing under uviol lamp, obtain fermentation medium, the ammoniacal liquor that is 2% with mass fraction regulates fermentation medium pH to 7.0; The fermentation medium of getting after the above-mentioned pH of 1.0L has regulated is put into 3L fermentation tank, by inoculum concentration 8%, mixed bacteria liquid is put into fermentation tank, and Temperature Setting is 30 DEG C, and speed setting is 150r/min, ferments 8 days, and described mixed bacteria liquid is that concentration is 1.5 × 10⁸Waterweed pseudomonad bacterium liquid and the concentration of individual/mL are 0.6 × 10⁸The bacillus pumilus liquid of individual/mL by volume 1:1 mixes; After fermentation ends, collect zymotic fluid, zymotic fluid is put into centrifuge, centrifugation 5min under 3000r/min, collects supernatant, and supernatant is put into beaker, in beaker, adds supernatant volume 30% absolute ethyl alcohol, stirs; After stirring, put into water-bath, with 5 DEG C/min speed, beaker is heated up, in the time that temperature rises to 35 DEG C, in beaker, add supernatant quality 2% compound protease, carry out insulated and stirred 1h with rotating speed 200r/min, and then continue to be warming up to 85 DEG C with 5 DEG C/min speed, after heating up, regulating pH with the potassium hydroxide solution of mass fraction 15% is 10.5, insulation 50min; After finishing, insulation naturally cools to room temperature, mixture in beaker is filtered with cellulose acetate filter membrane, collect filtrate and put into flask, to the absolute ethyl alcohol that adds 2 times of filtrate volumes in flask, add again filtrate quality 2% shitosan, be uniformly mixed, leave standstill until filter collection filtrate after producing without flocculate; The filtrate of gained is put into spray dryer sprays dry; collect dry thing and put into beaker; to the water that adds 70 DEG C in beaker; stir until dry thing stops adding water after dissolving completely; again to the 200 order carclazytes that add dry matter amount 6% in beaker, stir after 30min filtration under diminished pressure while hot, collect filtrate and put into spray dryer and spray and be dried; collect dry thing, can obtain high dispersive low-acyl gellan gum.

Take the gellan gum that 1g the present invention makes, be scattered in 250mL distilled water, be at room temperature stirred to abundant dissolving, in solution, add respectively 0.4g calcium chloride, 7g jelly powder, 120g white granulated sugar, 0.8g natrium citricum, 0.1g essence pigment, is heated to continuation at 60 DEG C and stirs 20min after stirring, after stirring, be cooled to room temperature, after cooling formation gel, can form jelly, at 100 DEG C, high-temperature sterilization is processed 10min.

Example 2

First meter by weight, get the sodium chloride solution of 35 parts of glucose, 15 parts of peptones, 2 parts of Dried Corn Steep Liquor Powder, 22 portions of beef extracts, 1.5 parts of dipotassium hydrogen phosphates, 1.5 parts of potassium dihydrogen phosphates and 23 parts of mass fractions 0.5%, after mixing, mixture is placed on to sterilizing under uviol lamp, obtain fermentation medium, the ammoniacal liquor that is 2% with mass fraction regulates fermentation medium pH to 7.0; The fermentation medium of getting after the above-mentioned pH of 1.3L has regulated is put into 3L fermentation tank, by inoculum concentration 9%, mixed bacteria liquid is put into fermentation tank, and Temperature Setting is 33 DEG C, and speed setting is 160r/min, ferments 9 days, and described mixed bacteria liquid is that concentration is 1.5 × 10⁸Waterweed pseudomonad bacterium liquid and the concentration of individual/mL are 0.6 × 10⁸The bacillus pumilus liquid of individual/mL by volume 1:1 mixes; After fermentation ends, collect zymotic fluid, zymotic fluid is put into centrifuge, centrifugation 6min under 4000r/min, collects supernatant, and supernatant is put into beaker, in beaker, adds supernatant volume 33% absolute ethyl alcohol, stirs; After stirring, put into water-bath, with 5 DEG C/min speed, beaker is heated up, in the time that temperature rises to 40 DEG C, in beaker, add supernatant quality 3% compound protease, carry out insulated and stirred 1.5h with rotating speed 200r/min, and then continue to be warming up to 90 DEG C with 5 DEG C/min speed, after heating up, regulating pH with the potassium hydroxide solution of mass fraction 15% is 10.8, insulation 55min; After finishing, insulation naturally cools to room temperature, mixture in beaker is filtered with cellulose acetate filter membrane, collect filtrate and put into flask, to the absolute ethyl alcohol that adds 2.5 times of filtrate volumes in flask, add again filtrate quality 2.5% shitosan, be uniformly mixed, leave standstill until filter collection filtrate after producing without flocculate; The filtrate of gained is put into spray dryer sprays dry; collect dry thing and put into beaker; to the water that adds 70 DEG C in beaker; stir until dry thing stops adding water after dissolving completely; again to the 200 order carclazytes that add dry matter amount 7% in beaker, stir after 35min filtration under diminished pressure while hot, collect filtrate and put into spray dryer and spray and be dried; collect dry thing, can obtain high dispersive low-acyl gellan gum.

Take the gellan gum that 1.5g the present invention makes, be scattered in 300mL distilled water, be at room temperature stirred to abundant dissolving, in solution, add respectively 0.5g calcium chloride, 8g jelly powder, 130g white granulated sugar, 1.0g natrium citricum, 0.15g essence pigment, is heated to continuation at 70 DEG C and stirs 25min after stirring, after stirring, be cooled to room temperature, after cooling formation gel, can form jelly, at 103 DEG C, high-temperature sterilization is processed 13min.

Example 3

First meter by weight, get the sodium chloride solution of 40 parts of glucose, 15 parts of peptones, 2 parts of Dried Corn Steep Liquor Powder, 18 portions of beef extracts, 2 parts of dipotassium hydrogen phosphates, 1 part of potassium dihydrogen phosphate and 22 parts of mass fractions 0.5%, after mixing, mixture is placed on to sterilizing under uviol lamp, obtain fermentation medium, the ammoniacal liquor that is 2% with mass fraction regulates fermentation medium pH to 7.0; The fermentation medium of getting after the above-mentioned pH of 1.5L has regulated is put into 3L fermentation tank, by inoculum concentration 10%, mixed bacteria liquid is put into fermentation tank, and Temperature Setting is 35 DEG C, and speed setting is 170r/min, ferments 10 days, and described mixed bacteria liquid is that concentration is 1.5 × 10⁸Waterweed pseudomonad bacterium liquid and the concentration of individual/mL are 0.6 × 10⁸The bacillus pumilus liquid of individual/mL by volume 1:1 mixes; After fermentation ends, collect zymotic fluid, zymotic fluid is put into centrifuge, centrifugation 8min under 5000r/min, collects supernatant, and supernatant is put into beaker, in beaker, adds supernatant volume 35% absolute ethyl alcohol, stirs; After stirring, put into water-bath, with 5 DEG C/min speed, beaker is heated up, in the time that temperature rises to 45 DEG C, in beaker, add supernatant quality 4% compound protease, carry out insulated and stirred 2h with rotating speed 200r/min, and then continue to be warming up to 95 DEG C with 5 DEG C/min speed, after heating up, regulating pH with the potassium hydroxide solution of mass fraction 15% is 11.0, insulation 60min; After finishing, insulation naturally cools to room temperature, mixture in beaker is filtered with cellulose acetate filter membrane, collect filtrate and put into flask, to the absolute ethyl alcohol that adds 3 times of filtrate volumes in flask, add again filtrate quality 3% shitosan, be uniformly mixed, leave standstill until filter collection filtrate after producing without flocculate; The filtrate of gained is put into spray dryer sprays dry; collect dry thing and put into beaker; to the water that adds 70 DEG C in beaker; stir until dry thing stops adding water after dissolving completely; again to the 200 order carclazytes that add dry matter amount 8% in beaker, stir after 40min filtration under diminished pressure while hot, collect filtrate and put into spray dryer and spray and be dried; collect dry thing, can obtain high dispersive low-acyl gellan gum.

Take the gellan gum that 2g the present invention makes, be scattered in 350mL distilled water, be at room temperature stirred to abundant dissolving, in solution, add respectively 0.6g calcium chloride, 9g jelly powder, 140g white granulated sugar, 1.2g natrium citricum, 0.2g essence pigment, is heated to continuation at 80 DEG C and stirs 30min after stirring, after stirring, be cooled to room temperature, after cooling formation gel, can form jelly, at 105 DEG C, high-temperature sterilization is processed 15min.

Claims (1)

1. a preparation method for high dispersive low-acyl gellan gum, is characterized in that concrete preparation process is

(1) count by weight, get the sodium chloride solution of 30～40 parts of glucose, 15～20 parts of peptones, 2～3 parts of Dried Corn Steep Liquor Powder, 18～22 portions of beef extracts, 1.5～2.2 parts of dipotassium hydrogen phosphates, 0.5～1.8 part of potassium dihydrogen phosphate and 21～23 parts of mass fractions 0.5%, after mixing, mixture is placed on to sterilizing under uviol lamp, obtain fermentation medium, the ammoniacal liquor that is 2% with mass fraction regulates fermentation medium pH to 7.0;

(2) fermentation medium of getting after the above-mentioned pH of 1.0～1.5L has regulated is put into 3L fermentation tank, by inoculum concentration 8～10%, mixed bacteria liquid is put into fermentation tank, Temperature Setting is 30～35 DEG C, speed setting is 150～170r/min, ferment 8～10 days, described mixed bacteria liquid is that concentration is 1.5 × 10⁸Waterweed pseudomonad bacterium liquid and the concentration of individual/mL are 0.6 × 10⁸The bacillus pumilus liquid of individual/mL by volume 1:1 mixes;

(3) after above-mentioned fermentation ends, collect zymotic fluid, zymotic fluid is put into centrifuge, centrifugation 5～8min under 3000～5000r/min, collect supernatant, supernatant is put into beaker, in beaker, add supernatant volume 30～35% absolute ethyl alcohols, stir;

(4) after above-mentioned stirring, put into water-bath, with 5 DEG C/min speed, beaker is heated up, in the time that temperature rises to 35～45 DEG C, in beaker, add supernatant quality 2～4% compound proteases, carry out insulated and stirred 1～2h with rotating speed 200r/min, and then continue to be warming up to 85～95 DEG C with 5 DEG C/min speed, after heating up, regulating pH with the potassium hydroxide solution of mass fraction 15% is 10.5～11.0, insulation 50～60min;

(5) after finishing, above-mentioned insulation naturally cools to room temperature, mixture in beaker is filtered with cellulose acetate filter membrane, collect filtrate and put into flask, to the absolute ethyl alcohol that adds 2～3 times of filtrate volumes in flask, add again filtrate quality 2～3% shitosans, be uniformly mixed, leave standstill until filter collection filtrate after producing without flocculate;

(6) filtrate of above-mentioned gained being put into spray dryer sprays dry; collect dry thing and put into beaker; to the water that adds 70 DEG C in beaker; stir until dry thing stops adding water after dissolving completely; again to the 200 order carclazytes that add dry matter amount 6～8% in beaker, stir after 30～40min filtration under diminished pressure while hot, collect filtrate and put into spray dryer and spray and be dried; collect dry thing, can obtain high dispersive low-acyl gellan gum.