CN116179626A - Method for fermenting and extracting low-acyl gellan gum - Google Patents
Method for fermenting and extracting low-acyl gellan gum Download PDFInfo
- Publication number
- CN116179626A CN116179626A CN202211498341.7A CN202211498341A CN116179626A CN 116179626 A CN116179626 A CN 116179626A CN 202211498341 A CN202211498341 A CN 202211498341A CN 116179626 A CN116179626 A CN 116179626A
- Authority
- CN
- China
- Prior art keywords
- gellan gum
- fermentation
- fermentation liquor
- precipitation
- acyl gellan
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229920002148 Gellan gum Polymers 0.000 title claims abstract description 58
- 239000000216 gellan gum Substances 0.000 title claims abstract description 58
- 235000010492 gellan gum Nutrition 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 36
- 238000000855 fermentation Methods 0.000 claims abstract description 54
- 230000004151 fermentation Effects 0.000 claims abstract description 54
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000001556 precipitation Methods 0.000 claims abstract description 22
- 150000003839 salts Chemical class 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims abstract description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 28
- 125000002252 acyl group Chemical group 0.000 claims description 14
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 12
- DVEKCXOJTLDBFE-UHFFFAOYSA-N n-dodecyl-n,n-dimethylglycinate Chemical compound CCCCCCCCCCCC[N+](C)(C)CC([O-])=O DVEKCXOJTLDBFE-UHFFFAOYSA-N 0.000 claims description 12
- 239000000047 product Substances 0.000 claims description 12
- 230000003311 flocculating effect Effects 0.000 claims description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 9
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 8
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 8
- 108091005658 Basic proteases Proteins 0.000 claims description 7
- 238000000108 ultra-filtration Methods 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 5
- 241000736110 Sphingomonas paucimobilis Species 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 241001052560 Thallis Species 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 230000003301 hydrolyzing effect Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 230000001105 regulatory effect Effects 0.000 claims description 5
- 239000005720 sucrose Substances 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 4
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 4
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 238000004042 decolorization Methods 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 238000000643 oven drying Methods 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 12
- 238000000605 extraction Methods 0.000 abstract description 7
- 230000008569 process Effects 0.000 description 6
- 239000003960 organic solvent Substances 0.000 description 5
- 238000001599 direct drying Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 230000020176 deacylation Effects 0.000 description 2
- 238000005947 deacylation reaction Methods 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000005189 flocculation Methods 0.000 description 2
- 230000016615 flocculation Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 150000004804 polysaccharides Chemical class 0.000 description 2
- 238000005185 salting out Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- RBNPOMFGQQGHHO-REOHCLBHSA-N L-glyceric acid Chemical group OC[C@H](O)C(O)=O RBNPOMFGQQGHHO-REOHCLBHSA-N 0.000 description 1
- 241000790234 Sphingomonas elodea Species 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 229920001586 anionic polysaccharide Polymers 0.000 description 1
- 150000004836 anionic polysaccharides Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- AEMOLEFTQBMNLQ-QIUUJYRFSA-N beta-D-glucuronic acid Chemical compound O[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-QIUUJYRFSA-N 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000004260 plant-type cell wall biogenesis Effects 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 150000004044 tetrasaccharides Chemical group 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Abstract
The invention provides a method for fermenting and extracting low-acyl gellan gum, which comprises the following steps: 1) Preparing fermentation liquor; 2) Pretreatment of fermentation liquor; 3) Salt precipitation; 4) Alcohol precipitation; 5) And separating and drying to obtain a low-acyl gellan gum finished product. The new technology research of extracting the low-acyl gellan gum by a salt-alcohol combination method mainly integrates the advantages of salt extraction and alcohol extraction, and fermentation liquor is subjected to salt precipitation and then alcohol precipitation, so that the product quality is improved, and the production cost is reduced.
Description
Technical Field
The invention relates to the technical field of biological fermentation, in particular to a method for fermenting and extracting low-acyl gellan gum.
Background
Gellan gum is a novel fully transparent gel, which is a linear anionic heteropolysaccharide. Is one of food glue integrating thickening, suspending, emulsifying and stabilizing, has excellent performance and has extremely high commercial profit and market prospect. The gellan gum has the advantages of low dosage, high transparency, strong fragrance releasing capability, acid resistance, alkali resistance and the like, and has wide application in food industry, medicine industry, daily chemical product industry and papermaking industry. Gellan gum is a polymeric anionic polysaccharide with a linear tetrasaccharide repeating unit as the main chain, and comprises beta-D-glucose, beta-D-glucuronic acid and alpha- (1-4) -L-rhamnose, wherein the composition ratio is about 2:1:1, the molecular weight is up to 500KD, and some groups are substituted by acetyl and L-glycerate.
At present, the process method for separating and extracting the low-acyl gellan gum product mainly comprises the following steps: organic solvent precipitation, salting-out precipitation, direct drying, ultrafiltration membrane separation, etc. But is limited by the respective conditions and characteristics in application. Different extraction methods have great influence on the quality and application characteristics of the low-acyl gellan gum product, and are also important factors for determining the cost of the product. Except for a direct drying method, the organic solvent precipitation method, the salting-out precipitation method, the ultrafiltration membrane separation method and the like all need washing and precipitation processes of the organic solvent. The solvents have strong volatility, and the original low-acyl gellan gum extraction process is batch or semi-closed, wherein a part of the solvents are mechanically and manually matched with open production. Therefore, the organic solvent consumption is large and accounts for 25% -40% of the production cost, and some production enterprises even reach more than 50% of the production cost. Moreover, these organic solvents are flammable and explosive dangerous materials, and the large amount of solvent evaporated will increase the risk of explosion and fire in the production site. Meanwhile, the intermittent and open production form has the advantages of large fluctuation of product quality, poor production environment, low efficiency and high labor intensity. Although the direct drying method and the enzyme-ultrafiltration-direct drying method can avoid the problems, the product contains more impurities and has poorer quality, and the application of the product is limited.
CN106350554a discloses a method for extracting non-transparent low acyl gellan gum, which comprises the following steps: step 1) fermentation, step 2) fermentation broth pretreatment, step 3) dehydration, step 4) alcohol dissolution, step 5) deacylation, step 6) flocculation, step 7) dehydration and pressing, step 8) crushing, and step 9) drying. The fermentation step adopts mixed fermentation of two strains, so that the fermentation efficiency is improved, but the mixed culture process parameters of the two strains are difficult to fix and normalize, and have larger uncertainty. Priority application number 2022105119276, filing date 2022.05.12.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for fermenting and extracting low-acyl gellan gum, which can effectively reduce the production cost of the low-acyl gellan gum and improve the product quality and yield.
The invention is realized by the following technical scheme.
A method for fermenting and extracting low acyl gellan gum comprising the steps of: 1) Preparing fermentation liquor; 2) Pretreatment of fermentation liquor; 3) Salt precipitation; 4) Alcohol precipitation; 5) And (5) separating and drying.
Specifically, the method comprises the following steps:
step 1) fermentation liquor preparation: inoculating Sphingomonas paucimobilis seed solution into a fermentation culture medium for fermentation culture for 60 hours to obtain fermentation liquor;
step 2) pretreatment of fermentation liquor: regulating pH value of the fermentation liquor to 10, and preserving heat at 70-80deg.C for 20-40min to obtain gellan gum liquid; adding CaCl 2 Flocculating and settling, filtering with a plate frame, removing thalli, diluting the filtrate for 2-10 times, adding alkaline protease, and hydrolyzing for 4 hours at the pH of 10 and 55 ℃; concentrating at 70-90deg.C by ultrafiltration for 2-6 times, decolorizing with active carbon, maintaining at 70-90deg.C for 20-60min, and removing activityCarbon, collecting decolorized solution;
step 3) salt precipitation: adding saturated sodium chloride solution accounting for 6% of the volume of the decolorized solution into the decolorized solution, precipitating and filtering to obtain a wet gellan gum product;
step 4) alcohol precipitation: adding 3-6 times of distilled water into wet gellan gum, stirring, adding 1-3 times of 70-80% ethanol, mixing, and flocculating low acyl gellan gum at 30-50deg.C;
step 5) separating and drying: standing to settle flocculate, discarding supernatant, collecting precipitate, and oven drying.
Preferably, the fermenter medium is added with hydrogen peroxide and dodecyl dimethyl betaine based on conventional fermentation medium.
More preferably, the components of the fermenter medium are: 40g/L of sucrose, 15g/L of corn steep liquor, 8g/L of ammonium sulfate, 1g/L of monopotassium phosphate, 1g/L of dipotassium phosphate, 0.8g/L of hydrogen peroxide, 0.4g/L of dodecyl dimethyl betaine, 0.1g/L of ferrous sulfate heptahydrate, 0.1g/L of citric acid and 0.1g/L of magnesium sulfate heptahydrate.
Preferably, in the fermentation culture process, dissolved oxygen is controlled to be 20%, the pressure is controlled to be 0.05Mpa, the stirring speed is controlled to be 200rpm, the pH is controlled to be 7.1, and the temperature is controlled to be 31 ℃.
Preferably, the step 2) pretreatment of the fermentation broth: regulating pH value of the fermentation liquor to 10, and preserving heat at 78 ℃ for 30min to obtain gellan gum liquid; adding 0.2% (w/v) CaCl 2 Flocculating and settling, filtering with a plate frame, removing thalli, diluting the filtrate for 6 times, adding 0.3% (w/v) alkaline protease, preserving the temperature for 4 hours at the pH of 10 and 55 ℃, and hydrolyzing protein; then, the mixture is concentrated by ultrafiltration at 80 ℃ for 4 times, then 2% (w/v) of active carbon is added for decolorization, the temperature is kept at 80 ℃ for 40min, the active carbon is removed, and decolorized solution is collected.
Preferably, the step 4) alcohol precipitation: adding 4 times of distilled water into wet gellan gum, stirring, adding 2 times of 70% ethanol, mixing, and flocculating low acyl gellan gum at 40deg.C.
More preferably, the alkaline protease has an enzyme activity of 10 ten thousand U/g.
The beneficial effects achieved by the invention mainly include, but are not limited to, the following:
the method for extracting the low acyl gellan gum, which is adopted by the invention, has the advantages that most impurities are removed by pretreatment of fermentation liquor, then the acyl is directly removed, the production cost of redissolving and deacylating the high acyl gellan gum is effectively reduced, the extraction yield of the low acyl gellan gum is effectively improved by the extraction method by a salinitol method, and the product quality is also improved.
In the fermentation process, the fermentation medium is optimized, and the yield of the gellan gum is greatly improved.
The extraction process is simple, the prices of the sodium chloride and the ethanol are low, the material cost can be reduced, and the method is suitable for large-scale industrial production.
The process adopts the pretreatment of the fermentation liquor and then directly deacylation treatment, thereby saving the cost of preparing the low-acyl gellan gum by adopting the high-acyl gellan gum and greatly reducing the production cost of the low-acyl gellan gum.
The invention uses alcohol in the alcohol precipitation flocculation low acyl gellan gum, the alcohol used in the process can be completely recovered, and the alcohol can enter the low acyl gellan gum production process again after the distillation reaches the required index, thereby realizing the repeated use of the alcohol.
Drawings
Fig. 1: influence of Hydrogen peroxide on gellan gum content in fermentation broth
Fig. 2: influence of dodecyl dimethyl betaine on gellan gum content of fermentation broth.
Detailed Description
In order that those skilled in the art will better understand the technical solutions of the present application, the present invention will be more clearly and completely described in the following in conjunction with specific embodiments of the present application, and it is apparent that the described embodiments are only some embodiments of the present application, not all embodiments. All other embodiments, which can be made by one of ordinary skill in the art based on the embodiments herein without making any inventive effort, shall fall within the scope of the present invention.
Example 1
A novel method for extracting low-acyl gellan gum from fermentation broth comprises the following specific steps:
step 1) fermentation liquor preparation: sphingomonas paucimobilis (Sphingomonas elodea) ATCC31461 was cultured to a concentration of 1X 10 8 Inoculating cfu/mL seed solution into a fermentation tank according to an inoculum size of 8% (volume ratio) for culturing for 60 hours to obtain fermentation liquor; in the fermentation process, controlling dissolved oxygen to be 20%, pressure to be 0.05Mpa, stirring speed to be 200rpm, pH to be 7.1 and temperature to be 31 ℃; maintaining the sugar concentration in the tank to be not lower than 2.0g/L by feeding 500g/L sucrose solution until the fermentation is finished; wherein, the components of the fermentation tank culture medium are as follows: 40g/L of sucrose, 15g/L of corn steep liquor, 8g/L of ammonium sulfate, 1g/L of monopotassium phosphate, 1g/L of dipotassium phosphate, 0.8g/L of hydrogen peroxide, 0.4g/L of dodecyl dimethyl betaine, 0.1g/L of ferrous sulfate heptahydrate, 0.1g/L of citric acid and 0.1g/L of magnesium sulfate heptahydrate; the solvent is water;
step 2) pretreatment of fermentation liquor: and (3) regulating the pH value of the fermentation liquor to 10, and preserving the temperature at 78 ℃ for 30min to obtain the low-acyl gellan gum liquid. Adding 0.2% (w/v) CaCl into low acyl gellan gum solution 2 Flocculating and settling, filtering with a plate frame, removing thalli, diluting the filtrate for 6 times, adding 0.3% (w/v) alkaline protease (10 ten thousand U/g), preserving the temperature for 4 hours at the pH of 10 and 55 ℃, and hydrolyzing protein; then, the mixture is concentrated by ultrafiltration at 80 ℃ for 4 times, then 2% (w/v) of active carbon is added for decolorization, the temperature is kept at 80 ℃ for 40min, the active carbon is removed, and decolorized solution is collected.
Step 3) salt precipitation: adding saturated sodium chloride solution accounting for 6% of the volume of the decolorized solution into the decolorized solution, precipitating, separating out and filtering to obtain a gellan gum wet product.
Step 4) alcohol precipitation: adding 4 times of distilled water into wet gellan gum, stirring, adding 2 times of 70% ethanol, mixing, and flocculating low acyl gellan gum at 40deg.C;
step 5) separating and drying: standing to settle flocculate, discarding supernatant, collecting precipitate, and oven drying to obtain the final product. The detection shows that the colloid purity can reach 92.6%, the yield is 83.1%, and the average molecular weight is maintained at about 73.8 kilodaltons.
Example 2
The fermentation medium optimizes the influence on the gum yield.
Conventional fermentation media: 40g/L of sucrose, 15g/L of corn steep liquor, 8g/L of ammonium sulfate, 1g/L of monopotassium phosphate, 1g/L of dipotassium phosphate, 0.1g/L of ferrous sulfate heptahydrate, 0.1g/L of citric acid and 0.1g/L of magnesium sulfate heptahydrate.
1. On the basis of a conventional fermentation medium, a certain amount of hydrogen peroxide is added, the adding amount is 0,0.2,0.4,0.8,1.6,3.2, the unit is g/L, as shown in fig. 1, the low-concentration hydrogen peroxide is positively correlated with the gellan gum yield, when the hydrogen peroxide concentration is increased to 0.8g/L, the gellan gum yield is 4.43g/L, and when the hydrogen peroxide concentration is increased to more than 1.6g/L, the gellan gum yield is reduced, probably because the damage to the strain can be caused by the overlarge hydrogen peroxide concentration, so that the yield of the gellan gum is reduced, and the low-concentration hydrogen peroxide can not only provide oxygen, but also play a certain stress role, and promote the synthesis of gellan gum extracellular polysaccharide.
2. On the basis of a conventional fermentation medium, a certain amount of dodecyl dimethyl betaine is added, the addition amount is 0,0.2,0.4,0.8,1.6,3.2, the unit is g/L, as shown in figure 2, the dodecyl dimethyl betaine has a certain promotion effect on gellan gum produced by Sphingomonas paucimobilis, but is weaker than hydrogen peroxide, the highest value is 4.01g/L, and after the concentration of the dodecyl dimethyl betaine is increased to 0.8g/L, the dodecyl dimethyl betaine has an obvious inhibition effect on the yield of the gellan gum,
dodecyl dimethyl betaine is a surfactant capable of promoting extracellular polysaccharide synthesis by inhibiting cell wall synthesis; however, if the inhibition of the cell wall is too strong, the activity of the cell is decreased, and thus the activity of the enzyme in the synthesis pathway is decreased, so that the gellan gum content is decreased.
3. Based on the test, 0.8g/L hydrogen peroxide and 0.4g/L dodecyl dimethyl betaine are added into a conventional fermentation medium. Under the same fermentation process conditions, the method has comparability, and the method discovers that the gellan gum produced by the Sphingomonas paucimobilis is greatly improved, and is increased by 1.57g/L and 43.1% compared with the conventional fermentation medium (3.64 g/L).
While the invention has been described with reference to specific embodiments thereof, it will be understood by those skilled in the art that the invention is not limited thereto, and that various changes and modifications may be made without departing from the spirit of the invention.
Claims (10)
1. A method for fermenting and extracting low acyl gellan gum comprising the steps of: 1) Preparing fermentation liquor; 2) Pretreatment of fermentation liquor; 3) Salt precipitation; 4) Alcohol precipitation; 5) And (5) separating and drying.
2. The method according to claim 1, characterized in that it comprises the steps of:
step 1) fermentation liquor preparation: inoculating Sphingomonas paucimobilis seed solution into a fermentation culture medium for fermentation culture for 60 hours to obtain fermentation liquor;
step 2) pretreatment of fermentation liquor: regulating pH value of the fermentation liquor to 10, and preserving heat at 70-80deg.C for 20-40min to obtain gellan gum liquid; adding CaCl 2 Flocculating and settling, filtering with a plate frame, removing thalli, diluting the filtrate for 2-10 times, adding alkaline protease, and hydrolyzing at pH10 and 55 ℃ for 4 hours; concentrating by ultrafiltration for 2-6 times, decolorizing with active carbon, maintaining at 70-90deg.C for 20-60min, removing active carbon, and collecting decolorized solution;
step 3) salt precipitation: adding saturated sodium chloride solution accounting for 6% of the volume of the decolorized solution into the decolorized solution, precipitating, and filtering to obtain a wet gellan gum product;
step 4) alcohol precipitation: adding 3-6 times of distilled water and 1-3 times of 70-80% ethanol into wet gellan gum, mixing, and flocculating low acyl gellan gum at 30-50deg.C;
step 5) separating and drying: standing to settle flocculate, discarding supernatant, collecting precipitate, and oven drying.
3. The method according to claim 2, wherein the fermenter medium is supplemented with hydrogen peroxide and dodecyl dimethyl betaine on the basis of a conventional fermentation medium.
4. The method of claim 2, wherein the components of the fermenter medium are: 40g/L of sucrose, 15g/L of corn steep liquor, 8g/L of ammonium sulfate, 1g/L of monopotassium phosphate, 1g/L of dipotassium phosphate, 0.8g/L of hydrogen peroxide, 0.4g/L of dodecyl dimethyl betaine, 0.1g/L of ferrous sulfate heptahydrate, 0.1g/L of citric acid and 0.1g/L of magnesium sulfate heptahydrate.
5. The method according to claim 2, wherein the dissolved oxygen is controlled to be 20%, the pressure is controlled to be 0.05Mpa, the stirring speed is 200rpm, the pH is controlled to be 7.1, and the temperature is controlled to be 31 ℃.
6. The method according to claim 2, wherein the step 2) fermentation broth pretreatment: regulating pH value of the fermentation liquor to 10, and preserving heat at 78 ℃ for 30min to obtain gellan gum liquid; adding 0.2% (w/v) CaCl 2 Flocculating and settling, filtering with a plate frame, removing thalli, diluting the filtrate for 6 times, adding 0.3% (w/v) alkaline protease, preserving the temperature for 4 hours at the pH of 10 and 55 ℃, and hydrolyzing protein; then, the mixture is concentrated by ultrafiltration at 80 ℃ for 4 times, then 2% (w/v) of active carbon is added for decolorization, the temperature is kept at 80 ℃ for 40min, the active carbon is removed, and decolorized solution is collected.
7. The method according to claim 2, wherein said step 4) alcohol precipitation: adding 4 times of distilled water and 2 times of 70% ethanol into wet gellan gum, mixing, and flocculating at 40deg.C.
8. The method according to claim 6, wherein the alkaline protease has an enzyme activity of 10U/g.
9. Gellan gum prepared according to the method of any of the preceding claims 1-8.
10. Use of gellan gum according to claim 9 in industry.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2022105119276 | 2022-05-12 | ||
| CN202210511927 | 2022-05-12 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116179626A true CN116179626A (en) | 2023-05-30 |
Family
ID=86446922
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211498341.7A Pending CN116179626A (en) | 2022-05-12 | 2022-11-28 | Method for fermenting and extracting low-acyl gellan gum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116179626A (en) |
-
2022
- 2022-11-28 CN CN202211498341.7A patent/CN116179626A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5765859B2 (en) | Yellow pigmentation defects Use in the production of sphingolipidoid aeromonas and gellan gum | |
| CN105695543B (en) | A kind of production method of surfactin | |
| CN101985641B (en) | Method for preparing bacterial cellulose by using wheat straw | |
| CN101020724A (en) | Process of preparing low molecular weight sodium hyaluronate | |
| CN104031956A (en) | Bacterial cellulose fermentation medium made from apple pomace and method for producing bacterial cellulose by utilizing medium | |
| CN104312834A (en) | Method for preparing sugarcane fruit wine | |
| CN102051395A (en) | Method for preparing bacterial cellulose from corn stalks | |
| CN112094752A (en) | Method for producing ultra-low molecular weight pullulan by fermenting aureobasidium pullulans | |
| CN113621674B (en) | Method for producing L-lactic acid by using liquor distiller grains | |
| CN108467876A (en) | A kind of fermentation process improving curdlan yield | |
| EP0319372B1 (en) | Process for the production of polysaccharides | |
| CN101985642B (en) | Method for preparing bacterial cellulose by using straw | |
| JPH0739386A (en) | Production of bacterial cellulose | |
| KR20020009480A (en) | Production of exopolysaccharides unattached to the surface of bacterial cells | |
| CN116179626A (en) | Method for fermenting and extracting low-acyl gellan gum | |
| DK165843B (en) | ENZYMATIC PROCEDURE FOR PREPARING A SYRUP WITH A HIGH CONTENT OF DEXTROSE | |
| CN106434443B (en) | A kind of production technology of Sodium Hyaluronate | |
| CN112608959B (en) | Method for improving fermentation unit of acetylglucosamine | |
| CN111286522B (en) | Preparation method of fermentation liquor containing rhamnolipid | |
| CN107674854B (en) | Nitrogen-fixing sphingosine monad and application thereof in preparation of gellan gum | |
| CN1124349C (en) | Process for preparing arabitol by transforming glucose with yeast cells | |
| CN112553259A (en) | Method for producing microbial flocculant by using Bacillus licheniformis | |
| CN102220394A (en) | Method for producing transparent xanthan gum | |
| CN105368912A (en) | Method for extracting sodium hyaluronate through spray reverse flow method | |
| CN117286082B (en) | Xanthomonas campestris and method for producing low-viscosity xanthan gum by fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |