CN101560537B - Fermentation method for producing high viscocity xanthan gum by xanthomonas campestris - Google Patents
Fermentation method for producing high viscocity xanthan gum by xanthomonas campestris Download PDFInfo
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a fermentation method for producing high viscocity xanthan gum by xanthomonas campestris, which relates to the technical field of the production of the xanthan gum by using microorganism fermentation engineering. The prior fermentation method for producing the xanthan gum by the xanthomonas campestris comprises: slant mother seeds, shaking culture, starter can culture, total culture medium fermentation culture and fermentation liquor extracting stage; and a fermentation culture stage of the invention is designed to be a basal culture medium fermentation culture stage which comprises a supplementary food culture medium A fed-batch, a food culture medium B fed-batch and a pH regulation and control process. Starch and crude farm product containing starchiness are sequentially batch-fed in two steps under the condition of firm control of fermentation liquor photodensity to increase the carbon nitrogen ratio gradually; the ferment pH value is controlled by an acid-base neutralization solution, so that the process fermentation time is reduced from 60 to 75 hours to 40 to 48 hours; high cost industrial raw materials are replaced by the low cost primary farm products, so that the raw material cost is reduced by 15 to 20 percent; the reduction of the fermentation time reduces the energy consumption by 25 to 30 percent; and experimental data shows that the feed stock conversion reaches 85 to 90 percent and the viscocity is improved by 10 to 20 percent.
Description
Technical field
The present invention relates to utilize microbial fermentation engineering production xanthan gum technical field, especially relate to a kind of fermentation process of producing high viscocity xanthan gum by xanthomonas campestris.
Background technology
Xanthan gum (Xanthan gum) claims yellow glue, xanthan gum again, is that xanthomonas campestris (Xanthomonascampestris) is main raw material with the carbohydrate, through a kind of broad-spectrum Microbial exopolysaccharides of fermentation engineering production.Nineteen fifty-two is separated to the cabbage black rot Xanthomonas campestris by the northern institute in USDA Yi Linuosi state skin Lille difficult to understand, and makes the wild cabbage extract be converted into the outer mixed polysaccharide of water miscible acid born of the same parents and obtain.Because this polysaccharide has very high viscosity, the thixotropy that flows and stable physico-chemical property, and it is nontoxic, pretend to additive and have vast market prospect in the detergents and cosmetic field, be collect thickening, suspension, emulsification in the world at present, be stable at one, xanthan gum that best performance is got over, be widely used in fields such as food, medicine, oil recovery, weaving, pottery, printing and dyeing, spices, makeup and fire-fighting.But, it is higher utilizing the production cost of microbial fermentation engineering production xanthan gum in the prior art, there are many deficiencies and defective, the xanthan gum production method that one is traditional, promptly utilize microbial fermentation, do fermention medium production xanthan gum with W-Gum or sucrose and peptone or corn steep liquor, the raw materials used macromolecular compounds such as polysaccharide and protein that are, cause being utilized slowly and the prolongation fermentation period by microorganism, domestic literature reports that its fermentation time is many at 60～75 hours, the long bigger consumption that causes naturally water, electricity, steam equal energy source of fermentation time; Increase the technology cost and partly adopt on-the-spot saccharification and hydrolysis method to degrade; Two, more traditional fermentation method adopts and is interrupted feed supplement or disposablely in substratum the nutritive ingredient preparation is finished, cause inoculum size big and must three grades or more multistage fermentation, insufficient energy utilization rate that causes of conducting heat is low, does not utilize raw material to increase the reduction that the fermented liquid viscosity causes the oxygen transfer efficiency; Three, because the full-bodied characteristic of xanthan gum product and fermentation itself is the process of a high oxygen consumption, later stage technology severe depletion of oxygen causes fermenting, the viscosity performance that fermentation time is got over long products can descend gradually, for example the coming off and lack of side-chain radical is unfavorable more to obtaining high-quality xanthan gum product.The researchist is devoted to make great efforts to reduce its production cost by the whole bag of tricks always for a long time, for example Chinese patent application number is that 200810137040.5 patent documentation discloses a kind of Xanthomonas campestris polysaccharide substratum and preparation method thereof, and characteristics are to utilize useless starch water to do substratum.A kind of production technique of utilizing the beet sirup fermenting and producing xanthan gum etc. is disclosed and China Patent No. is the patent documentation of ZL03124005.4.This research is then studied its fermention medium and fermentation process thereof at the problems referred to above that exist in the traditional processing technology and is innovated, and has proposed a kind of fermentation process of producing high viscocity xanthan gum by xanthomonas campestris.
Summary of the invention
The technical problem to be solved in the present invention is exactly the fermentation process that discloses a kind of producing high viscocity xanthan gum by xanthomonas campestris, to overcome deficiency and the defective that prior art exists in fermentation period, raw materials cost and energy consumption, reach the purpose that shortens fermentation period, reduction raw materials cost, reduces energy consumption.
The purpose technical solution that realizes the present invention is as follows:
A kind of fermentation process of producing high viscocity xanthan gum by xanthomonas campestris, comprise the female kind in inclined-plane, shake-flask culture, seed tank culture, full substratum fermentation culture, broth extraction stage, it is characterized in that: full substratum fermentation culture stage is designed to the basic medium fermentation culture stage and comprises that supplemented medium A stream adds, supplemented medium B stream adds, PH regulates and control link, and concrete steps are as follows:
(1) can utilize carbon, nitrogenous source substratum such as monose and protolysate fully fast according to Xanthomonas campestris, principle as fermention medium fast breeding Xanthomonas campestris, at first prepare basic medium, its basic medium component (g/L weightmeasurement ratio) be prepared as:
(2) the Xanthomonas campestris liquid after the cascade amplification culture (bacterial classification CICC10258) is seeded in the basic fermention medium, inoculum size 10% (volume ratio), the pH value is controlled at 6.5～7.5,30 ℃ of temperature, fermentation time continues 8～12 hours;
The component of supplemented medium A and supplemented medium B (g/L weightmeasurement ratio) be prepared as:
A, supplemented medium A
W-Gum 120, Dried Corn Steep Liquor Powder 60, citric acid 2;
B, supplemented medium B
Semen Maydis powder 300, citric acid 2;
Culture temperature: 30 ℃, cultivate pH value 6.5～7.5;
Training method: oxygen consumption is cultivated;
Substratum adopts said components, and equal 121 ℃, the 20min preparation of sterilizing;
(3) add supplemented medium A and supplemented medium B with the mode of flow feeding in basic fermention medium by given pace, breeding optical density(OD) at its fermented liquid thalline is under OD60010.0～12.0 conditions the disposable benefit of supplemented medium A to be gone in the fermented liquid, and it is earlier fermentation that fermentation time continues 6～8 hours; After supplemented medium A finishes by above step feed supplement, promptly use supplemented medium B to begin flow feeding, flow feeding was gone into the disposable benefit of supplemented medium B in the fermented liquid after 16～20 hours fast, simultaneously fermentating liquid PH value is adjusted to 4.5～5.5 and cultivates, fermentation time continues 6～8 hours latter stages of promptly fermenting and finishes fermentation.
The fermentation process of described a kind of producing high viscocity xanthan gum by xanthomonas campestris is characterized in that, the feed supplement fed-batch mode adds by given pace stream and is meant: the feed supplement initial rate of supplemented medium A adds by 30～60L/hr stream respectively; The feed rate of supplemented medium B adds by 40～50L/hr stream.
The fermentation process of described a kind of producing high viscocity xanthan gum by xanthomonas campestris is characterised in that, the total OD600 of fermented liquid optical density(OD) is controlled at 13.0～15.0 in the fermenting process; When total OD600 adds to basic fermention medium supplemented medium A is disposable 10.0～12.0 the time.
The fermentation process of described a kind of producing high viscocity xanthan gum by xanthomonas campestris is characterised in that, described fermenting process mid-early stage fermentation pH value is controlled in 6.5～7.5 the scope, and fermenting is controlled in 4.5～5.5 scopes latter stage.
The fermentation process of described a kind of producing high viscocity xanthan gum by xanthomonas campestris is characterized in that the fermentation time periodic Control is about 40～48 hours.
Described carbon source comprise glucose, sucrose, maltose, W-Gum, potato starch, Semen Maydis powder, Tapioca Starch, potato dry powder etc. one or more; Described nitrogenous source comprise ammonium sulfate, ammonium chloride, urea, hydrolytic soya bean protein peptone, extractum carnis, yeast extract paste, Dried Corn Steep Liquor Powder etc. one or more.
Compared with prior art, advantage and beneficial effect thereof that the present invention produces are: its gordian technique is the Xanthomonas campestris that cascade is amplified to be inserted contain in the carbon nitrogen source substratum that monose and protolysate etc. can utilize fast by microorganism, under the situation of strictness control fermented liquid optical density(OD) successively in two steps stream add starch and contain the thick agricultural-food of starchiness, progressively increase carbon-nitrogen ratio, by acid-base neutralisation hydraulic control system fermentation pH value, reached with the technology fermentation time foreshortened to 40-48 hour from 60-75 hour, fermentation time shortening about 30%; Being improved to cheaply expensive industrial raw material just, agricultural-food substitute, raw materials cost reduces by 15～20%, the saving sterilisation vapour caused energy consumption to reduce by 25～30% after fermentation time shortened back saves energy, technology adjustment, show by test for data, feed stock conversion reaches 85～90%, and viscosity improves 10～20%.
Description of drawings
Fig. 1 is each processing step schematic block diagram of traditional zymotic method of the present invention.
Fig. 2 is each the processing step schematic block diagram of fermentation method after the improvement of the present invention.
Fig. 3 is the half sponge process product application performance comparison chart after the improvement of the present invention.
Embodiment
With embodiment the present invention is done further in conjunction with the accompanying drawings and specify.As can be seen from Figure 1, traditional fermentation method comprises the female kind in inclined-plane, shake-flask culture, seed tank culture, secondary seed cultivation, full substratum fermentation culture, broth extraction stage.As can be seen from Figure 2, each processing step of fermentation method after the improvement of the present invention is at full substratum fermentation culture stage in the traditional fermentation method of Fig. 1, the present invention has carried out innovative design, and this stage is set to the basic medium fermentation culture stage and comprises each sport technique segment step such as supplemented medium A stream adds, supplemented medium B stream adds, PH regulation and control.Concrete steps comprise: at first prepare basic fermention medium and begin fermentation, its basic fermention medium component (g/L weightmeasurement ratio) is glucose 10; Hydrolytic soya bean protein peptone 1; Urea 2, Dried Corn Steep Liquor Powder 3; K
2HPO
45, citric acid 1, MgSO
47H
2O 0.3, CaCO
33, sterilization back NaOH transfers PH to 6.5～7.5, coefficient 0.6; Secondly, with the bacterial classification after the cascade amplification culture number be the Xanthomonas campestris liquid inoculation of CICC10258 to basic fermention medium, inoculum size 10% (volume ratio), the pH value is controlled at 6.5～7.5,30 ℃ of temperature, lasting 8～12 hours of fermentation time; They are three years old, the component of supplemented medium A and supplemented medium B is important, feed stock conversion by evidence W-Gum and Dried Corn Steep Liquor Powder is respectively 88% and 87%, has therefore selected the feed composition as supplemented medium such as W-Gum and Dried Corn Steep Liquor Powder for use, and it is prepared as follows:
A, supplemented medium A (g/L weightmeasurement ratio)
W-Gum 120, Dried Corn Steep Liquor Powder 60, citric acid 2;
B, supplemented medium B
Semen Maydis powder 300, citric acid 2;
Culture temperature: 30 ℃, cultivate pH value 6.5～7.5;
Training method: oxygen consumption is cultivated.
Four, get hold of every step flow feeding mode.After needing to add by given pace stream earlier again property add; Given pace stream adds and is meant: the feed supplement initial rate of supplemented medium A is pressed 30L/hr respectively, 40L/hr, and 50L/hr, 60L/hr stream adds, and 40L/hr is constant for supplemented medium B feed rate; The feed rate of supplemented medium B is pressed 30L/hr respectively, 40L/hr, and 50L/hr, 60L/hr stream adds, and 50L/hr is constant for supplemented medium A feed rate.The method that adds with stream is added supplemented medium A and supplemented medium B in basic fermention medium by given pace, breeding optical density(OD) at its fermented liquid thalline is under OD60010.0～12.0 conditions the disposable benefit of supplemented medium A to be gone in the fermented liquid, and fermentation time continues 6～8 hours; After supplemented medium A finishes by above step feed supplement, promptly use supplemented medium B to begin flow feeding, flow feeding was gone into the disposable benefit of supplemented medium B in the fermented liquid after 16～20 hours fast, simultaneously fermentating liquid PH value being adjusted to 4.5～5.5 cultivates, fermentation time continues to finish fermentation, 40～48 hours whole cycles after 6～8 hours.Its five, to be controlled at 13.0～15.0 also be important to the total OD600 of fermented liquid optical density(OD) in the fermenting process; When total OD600 adds to basic fermention medium supplemented medium A is disposable 10.0～12.0 the time.In its six fermenting process, earlier fermentation pH value is controlled in 6.5～7.5 the scope, and fermentation is controlled in 4.5～5.5 scopes latter stage.The fermentation time periodic Control is about 40～48 hours.Fermentation result such as following table: (table 1)
| Batch | Incubation time | Culture transferring OD600 | OD600 is changed in feed supplement | Feed stock conversion | 6r/min viscosity (mPaS) | Produce glue yield |
| 1 | 44 | 4.23 | 10.82 | 89.7％ | 33100 | 5.38％ |
| 2 | 40 | 4.18 | 10.88 | 87.0％ | 34300 | 5.22％ |
| 3 | 44 | 4.21 | 10.75 | 88.6％ | 33400 | 5.32％ |
Fig. 3 is the half sponge process product application performance comparison chart after the improvement of the present invention, can prove the beneficial effect that the present invention produces.
Described basic fermention medium required carbon source comprise glucose, sucrose, maltose, W-Gum, potato starch, Semen Maydis powder, Tapioca Starch, potato dry powder etc. one or more can select to use.Fermention medium required nitrogenous source in basis comprises ammonium sulfate, ammonium chloride, urea, hydrolytic soya bean protein peptone, extractum carnis, yeast extract paste, Dried Corn Steep Liquor Powder etc., and one or more can select to use.Soda acid PH conditioning agent comprises NaOH, lime carbonate, calcium oxide, sulfuric acid, phosphoric acid etc., and one or more can select to use.From following table 2 as can be known the method for the invention with compare with starch or sucrose fermentation method merely, raw materials cost reduces by 15～20%, energy consumption reduces by 25～30%, viscosity improves about 10～20%.
Different material is to the influence (table 2) of xanthan gum output
| Material composition | Culture transferring OD600 | OD600 is changed in feed supplement | Incubation time | 6r/min viscosity (mPaS) | Produce glue rate % | Feed stock conversion |
| Beef peptone | 4.15 | 10.86 | 44 | 29300 | 4.91 | 81.8 |
| The hydrolytic soya bean protein peptone | 4.37 | 11.73 | 43 | 32200 | 5.20 | 86.7 |
| Soybean cake powder | 4.12 | 10.62 | 46 | 33400 | 5.13 | 85.5 |
| Dried Corn Steep Liquor Powder | 4.02 | 10.54 | 50 | 33900 | 5.09 | 84.8 |
| Glucose | 4.30 | 11.53 | 48 | 29700 | 4.96 | 82.7 |
| Sucrose | 4.21 | 10.91 | 46 | 29200 | 4.97 | 82.8 |
| W-Gum | 3.88 | 10.27 | 48 | 28800 | 4.88 | 81.3 |
Embodiment 1
The present invention adopts the second order fermentation method
Shake-flask culture: preparation shake-flask culture base 3L, shake-flask culture base component (g/L envelope-bulk to weight ratio):
PH is adjusted to 7.0,6 parts of packing, and sterilization respectively meets 1ml glycerine plumber and makes seed on the super clean bench, 30 ℃, cultivated in 16 hours;
Seed tank culture: seed culture medium component (g/L envelope-bulk to weight ratio):
Preparation seed culture medium 600L is in the 1000L fermentor tank, and PH is adjusted to 7.0, and sterilization inserts shake-flask seed liquid, stirs 150 ~ 250 commentaries on classics/min, ventilation 0.3vvm, and 30 ℃, cultivated in 8 ~ 10 hours, total OD is to 4.03 culture transferrings;
Fermentation culture: basic fermention medium component (g/L envelope-bulk to weight ratio):
Supplemented medium A component (g/L envelope-bulk to weight ratio):
W-Gum 120, Dried Corn Steep Liquor Powder 60, citric acid 2;
Supplemented medium B component (g/L envelope-bulk to weight ratio):
Semen Maydis powder 300, citric acid 2;
Culture temperature: 30 ℃, cultivate pH value 6.5～7.5;
Training method: oxygen consumption is cultivated.
Substratum adopts said components, and equal 121 ℃, the 20min preparation of sterilizing;
Preparation supplemented medium A1 ton is in 2 600L feed supplement jars, and 5% bubble opposes in 50L bubble enemy jar, and the soda acid ullage disappears, sterilization, the seed liquor culture transferring to sterilization fermentation liquid, 100～150 commentariess on classics/min, the 0.3～0.6vvm that ventilates, PH is adjusted to 6.5,30 ℃ of cultivations; Press beginning feed supplement in 30L/ hour simultaneously, double every 4 hours feed rates, and sampling detection OD600, the total OD of fermenting process is controlled at 13.0～15.0; When total OD 10.0 the time, will remain supplemented medium A and once mend in the fermented liquid,
Preparation supplemented medium B1 ton, it is stand-by to sterilize; Begin by feed supplement again in 40L/ hour after last step feed supplement finishes, will remain after 16～20 hours feed supplement time in the disposable adding fermented liquid of feed supplement, simultaneously PH is controlled at 4.5 to fermentation ends, the fermentation time periodic Control is about 44 hours.
Embodiment 2
Except that the Variation order VO factor, other composition of substratum remains unchanged, and equal 121 ℃, the 20min preparation of sterilizing;
Shake-flask culture: preparation shake-flask culture base 3L, pH regulator to 7.0,6 parts of packing, sterilization respectively meets 1ml glycerine plumber and makes seed on the super clean bench, and 30 ℃, 16hr cultivates;
Seed culture: preparation seed culture medium 600L is in the 1000L fermentor tank, and pH regulator to 7.0 is sterilized, and inserts shake-flask seed liquid, stirs 150 ~ 250 commentaries on classics/min, ventilation 0.3vvm, and 30 ℃, 8 ~ 10hr cultivates, and always OD is to 4.21 culture transferrings;
Fermentation culture: 6 tons of fermention mediums of preparation are in 10 tons of fermentor tanks, preparation supplemented medium A1 ton is in 2 600L feed supplement jars, 5% bubble opposes in 50L bubble enemy jar, the soda acid ullage disappears, sterilization, and the seed liquor culture transferring is to the sterilization fermentation nutrient solution, 100～150 commentaries on classics/min, ventilation 0.3～0.6vvm, pH regulator to 6.9,30 ℃ of cultivations; Begin feed supplement by 40L/hr simultaneously, double every the 4hr feed rate, and sampling detection OD600, the total OD of fermenting process is controlled at 13.0 ~ 15.0; When total OD 10.93 the time, will remain supplemented medium A and once mend in the fermented liquid, and preparation supplemented medium B1 ton, it is stand-by to sterilize; Begin the feed supplement by 40L/hr after last step feed supplement finishes, will remain behind feed supplement time 16 ~ 20hr in the disposable adding fermented liquid of feed supplement, simultaneously pH is controlled at 4.9 to fermentation ends, the fermentation time periodic Control is about 40 hours.
Embodiment 3
Except that the Variation order VO factor, other composition of substratum remains unchanged, and equal 121 ℃, the 20min preparation of sterilizing;
Shake-flask culture: preparation shake-flask culture base 3L, pH regulator to 7.0,6 parts of packing, sterilization respectively meets 1ml glycerine plumber and makes seed on the super clean bench, and 30 ℃, 16hr cultivates;
Seed culture: preparation seed culture medium 600L is in the 1000L fermentor tank, and pH regulator to 7.0 is sterilized, and inserts shake-flask seed liquid, stirs 150 ~ 250 commentaries on classics/min, ventilation 0.3vvm, and 30 ℃, 8 ~ 10hr cultivates, and always OD is to 4.18 culture transferrings;
Fermentation culture: 6 tons of fermention mediums of preparation are in 10 tons of fermentor tanks, preparation supplemented medium A1 ton is in 2 600L feed supplement jars, 5% bubble opposes in 50L bubble enemy jar, the soda acid ullage disappears, sterilization, and the seed liquor culture transferring is to the sterilization fermentation nutrient solution, 100 ~ 150 commentaries on classics/min, ventilation 0.3 ~ 0.6vvm, pH regulator to 7.2,30 ℃ of cultivations; Begin feed supplement by 50L/hr simultaneously, double every the 4hr feed rate, and sampling detection OD600, the total OD of fermenting process is controlled at 13.0～15.0; When total OD 11.54 the time, will remain supplemented medium A and once mend in the fermented liquid, and preparation supplemented medium B1 ton, it is stand-by to sterilize; Begin the feed supplement by 40L/hr after last step feed supplement finishes, will remain behind feed supplement time 16～20hr in the disposable adding fermented liquid of feed supplement, simultaneously pH is controlled at 5.2 to fermentation ends, the fermentation time periodic Control is about 45 hours.
Embodiment 4
Except that the Variation order VO factor, other composition of substratum remains unchanged, and equal 121 ℃, the 20min preparation of sterilizing;
Shake-flask culture: preparation shake-flask culture base 3L, pH regulator to 7.0,6 parts of packing, sterilization respectively meets 1ml glycerine plumber and makes seed on the super clean bench, and 30 ℃, 16hr cultivates;
Seed culture: preparation seed culture medium 600L is in the 1000L fermentor tank, and pH regulator to 7.0 is sterilized, and inserts shake-flask seed liquid, stirs 150 ~ 250 commentaries on classics/min, ventilation 0.3vvm, and 30 ℃, 8 ~ 10hr cultivates, and always OD is to 4.33 culture transferrings;
Fermentation culture: 6 tons of fermention mediums of preparation are in 10 tons of fermentor tanks, preparation supplemented medium A1 ton is in 2 600L feed supplement jars, 5% bubble opposes in 50L bubble enemy jar, the soda acid ullage disappears, sterilization, and the seed liquor culture transferring is to the sterilization fermentation nutrient solution, 100 ~ 150 commentaries on classics/min, ventilation 0.3 ~ 0.6vvm, pH regulator to 7.5,30 ℃ of cultivations; Begin feed supplement by 60L/hr simultaneously, double every the 4hr feed rate, and sampling detection OD600, the total OD of fermenting process is controlled at 13.0～15.0; When total OD 12.0 the time, will remain supplemented medium A and once mend in the fermented liquid, and preparation supplemented medium B1 ton, it is stand-by to sterilize; Begin the feed supplement by 40L/hr after last step feed supplement finishes, will remain behind feed supplement time 16～20hr in the disposable adding fermented liquid of feed supplement, simultaneously pH is controlled at 5.5 to fermentation ends, the fermentation time periodic Control is about 48 hours.
Most preferred embodiment:
Each substratum is in preparation of the ratio in claims and sterilization;
Shake-flask culture: preparation shake-flask culture base 3L, pH regulator to 7.0,6 parts of packing, sterilization respectively meets 1ml glycerine plumber and makes seed on the super clean bench, and 30 ℃, 16h cultivates;
Seed culture: preparation seed culture medium 600L is in the 1000L fermentor tank, and pH regulator to 7.0 is sterilized, and inserts shake-flask seed liquid, stirs 150～250 commentaries on classics/min, ventilation 0.3vvm, and 30 ℃, 10h cultivates, and always OD is to 4.2 culture transferrings;
Fermentation culture: 6 tons of fermention mediums of preparation are in 10 tons of fermentor tanks, preparation 1) 1 ton of supplemented medium is in 2 600L feed supplement jars, 5% bubble opposes in 50L bubble enemy jar, the soda acid ullage disappears, sterilization, and the seed liquor culture transferring is to sterilization fermentation liquid, 100～150 commentaries on classics/min, ventilation 0.3～0.6vvm, pH regulator to 7 ± 0.5,30 ℃ cultivation; Begin to add supplemented medium A by 40L/h simultaneously, double every the 4h feed rate, and sampling detection OD600, the total OD of fermenting process is controlled at 13.0～15.0; When total OD 10.80 the time, will remain supplemented medium A and once mend in the fermented liquid, and preparation supplemented medium B1 ton, it is stand-by to sterilize; After last step feed supplement finishes, begin feed supplement, will remain behind the feed supplement time 16hr in the disposable adding fermented liquid of feed supplement, simultaneously pH is controlled at 5.0 ± 0.2, finish fermentation behind the 8hr by 50L/hr.The fermenting process data are as follows:
The research trial embodiment data that provide the present invention below and done:
1, different supplemented mediums are to the influence test of xanthan gum output
Except that supplemented medium, carbon source adopts glucose in all the other substratum, and nitrogenous source adopts the hydrolytic soya bean protein peptone, and all the other compositions remain unchanged, and when supplemented medium A composition changed, supplemented medium B was constant, and vice versa, and fermentation technology is the same;
1) supplemented medium A carbon source adopts sucrose, W-Gum, Semen Maydis powder respectively, and nitrogenous source adopts Dried Corn Steep Liquor Powder, and supplemented medium B uses sucrose as carbon source;
2) supplemented medium B carbon source adopts sucrose, W-Gum, Semen Maydis powder respectively; Supplemented medium A carbon source W-Gum, the nitrogenous source Dried Corn Steep Liquor Powder.
Fermentation result such as following table:
Test 1)
| Material composition | Culture transferring OD600 | OD600 is changed in feed supplement | Incubation time | 6r/min viscosity (mPaS) | Produce glue rate % | Feed stock conversion % |
| Sucrose | 4.38 | 11.64 | 42 | 32800 | 5.33 | 88.8 |
| W-Gum | 4.23 | 10.41 | 48 | 31600 | 4.82 | 80.3 |
| Semen Maydis powder | 4.42 | 9.96 | 58 | 29800 | 4.61 | 76.8 |
Test 2):
| Material composition | Culture transferring OD600 | OD600 is changed in feed supplement | Incubation time | 6r/min viscosity (mPaS) | Produce glue rate % | Feed stock conversion % |
| Sucrose | 4.27 | 11.64 | 42 | 34700 | 5.42 | 90.3 |
| W-Gum | 4.31 | 10.93 | 45 | 33900 | 5.28 | 88.0 |
| Semen Maydis powder | 4.52 | 11.50 | 46 | 33600 | 5.22 | 87.0 |
2, different feed rates are to the influence test of xanthan gum output
Experimental design: fermentation technology is constant, and culture medium prescription is filled a prescription by described in claims;
1) the feed supplement initial rate of supplemented medium A is pressed 30L/hr respectively, 40L/hr, and 50L/hr, 60L/hr, 40L/hr is constant for supplemented medium B feed rate;
2) feed rate of supplemented medium B is pressed 30L/hr respectively, 40L/hr, and 50L/hr, 60L/hr, 50L/hr is constant for supplemented medium A feed rate;
The result is as follows in fermentation:
Test 1):
| Feed rate | Culture transferring OD600 | OD600 is changed in feed supplement | Incubation time | 6r/min viscosity (mPaS) | Produce glue rate % | Feed stock conversion % |
| 30L/hr | 4.03 | 10.69 | 46 | 30700 | 4.97 | 82.8 |
| 40L/hr | 4.21 | 10.93 | 46 | 33500 | 5.16 | 86.0 |
| 50L/hr | 4.18 | 11.50 | 46 | 33100 | 5.23 | 87.2 |
| 60L/hr | 4.33 | 10.35 | 46 | 34400 | 5.12 | 85.3 |
Test 2):
| Feed rate | Culture transferring OD600 | OD600 when feed supplement is changed | Incubation time | 6r/min viscosity (mPaS) | Produce glue rate % | Feed stock conversion % |
| 30L/hr | 4.03 | 10.69 | 46 | 31400 | 5.08 | 84.6 |
| 40L/hr | 4.21 | 11.53 | 46 | 33800 | 5.13 | 85.5 |
| 50L/hr | 4.18 | 10.77 | 46 | 34200 | 5.26 | 87.7 |
| 60L/hr | 4.33 | 11.25 | 46 | 34500 | 5.19 | 86.5 |
3, the influence of cell concentration to xanthan gum output changed in feed supplement
Experimental design: fermentation technology is constant, and culture medium prescription is filled a prescription by described in claims; The total OD600 of fermented liquid when supplemented medium A is replaced by B is respectively 9.0,10.5,11.5,13.0 and carries out fermenting experiment;
The result is as follows in fermentation:
| OD600 when feed supplement is changed | Culture transferring OD600 | Incubation time | 6r/min viscosity (mPaS) | Produce glue rate % | Feed stock conversion % |
| 9.52 | ?4.28 | 46 | 31400 | 5.07 | 84.5 |
| 10.48 | ?4.13 | 46 | 34200 | 5.23 | 87.2 |
| 11.54 | ?4.22 | 46 | 33900 | 5.17 | 86.2 |
| 13.03 | ?4.36 | 46 | 29800 | 4.98 | 83.0 |
4, the xanthan gum performance comparison situation of the present invention and other manufacturer production
A: the xanthan gum that the method for the invention is produced, B, C: domestic other two manufacturer production ground xanthan gum; By same ratio and other compositions are composite detects, data are as follows with three kinds of xanthan gum:
Claims (2)
1. the fermentation process of a producing high viscocity xanthan gum by xanthomonas campestris, comprise the female kind in inclined-plane, shake-flask culture, seed tank culture, full substratum fermentation culture, broth extraction stage, it is characterized in that: full substratum fermentation culture stage is designed to the basic medium fermentation culture stage, comprise that supplemented medium A stream adds, supplemented medium B stream adds, pH regulates and control link, concrete steps are as follows:
(1) can utilize carbon, nitrogenous source substratum such as monose and protolysate fully fast according to Xanthomonas campestris, principle as fermention medium fast breeding Xanthomonas campestris, at first prepare basic fermention medium, its basic fermention medium component is represented with g/L and is prepared as:
Glucose 10; Hydrolytic soya bean protein peptone 1; Urea 2, Dried Corn Steep Liquor Powder 3; K
2HPO
45, citric acid 1, MgSO
47H
2O 0.3, CaCO
33,121 ℃, the 20min sterilization, sterilization back NaOH transfers pH to 6.5～7.5, coefficient 0.6;
(2) with the Xanthomonas campestris liquid inoculation of the bacterial classification CICC10258 after the cascade amplification culture to basic fermention medium, by 10% volume ratio inoculation, pH value is controlled at 6.5～7.5,30 ℃ of temperature, lasting 8～12 hours of fermentation time;
The component of supplemented medium A and supplemented medium B is represented with g/L and is prepared as:
A, supplemented medium A
W-Gum 120, Dried Corn Steep Liquor Powder 60, citric acid 2;
B, supplemented medium B
Semen Maydis powder 300, citric acid 2;
Training method: oxygen consumption is cultivated;
Substratum adopts said components, and equal 121 ℃, the 20min preparation of sterilizing;
(3) mode that adds with feed supplement stream is added supplemented medium A and supplemented medium B in basic fermention medium, and the feed supplement initial rate of supplemented medium A adds by 30～60L/hr stream; Breeding optical density(OD) at its fermented liquid thalline is OD
600Under 10.0～12.0 conditions the disposable benefit of supplemented medium A is gone in the fermented liquid, fermentation time continues 6～8 hours, and pH is controlled in the scope of 6.5-7.5; After supplemented medium A finishes by above step feed supplement, promptly use supplemented medium B to begin flow feeding, the feed rate of supplemented medium B adds by 40～50L/hr stream, flow feeding was gone into the disposable benefit of supplemented medium B in the fermented liquid after 16～20 hours fast, fermented liquid pH value is adjusted to 4.5～5.5 cultivates, fermentation time continues 6～8 hours, and the pH value is controlled in 4.5～5.5 the scope, finishes fermentation.
2. the fermentation process of a kind of producing high viscocity xanthan gum by xanthomonas campestris according to claim 1 is characterized in that the fermentation time periodic Control was at 40～48 hours.
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| CN101906390B (en) * | 2010-03-25 | 2012-05-02 | 淄博中轩生化有限公司 | Xanthomonas, preparation method thereof and method thereof for producing temperature resistant xanthan gum polysaccharide |
| CN101838625B (en) * | 2010-04-02 | 2012-07-11 | 淄博中轩生化有限公司 | Xanthomonas campestris and method for preparing xanthan gum specially used for soy sauce thereby |
| CN102220394A (en) * | 2010-04-16 | 2011-10-19 | 淄博中轩生化有限公司 | Method for producing transparent xanthan gum |
| CN103205471B (en) * | 2013-04-26 | 2014-07-09 | 新疆阜丰生物科技有限公司 | Xanthan gum preparation method suitable for high-concentration multivalent ion liquid |
| CN107287260B (en) * | 2016-04-11 | 2020-09-15 | 新疆梅花氨基酸有限责任公司 | Production method of low-cost high-quality xanthan gum |
| CN106674342B (en) * | 2017-02-24 | 2020-05-22 | 山东阜丰发酵有限公司 | Extraction process of corn gluten protein and application of corn gluten protein in xanthan gum fermentation |
| CN110551787A (en) * | 2018-05-31 | 2019-12-10 | 卢松 | Process for optimizing fermentation of xanthan gum bacteria by using zymolytic protein |
| CN109929893A (en) * | 2019-04-12 | 2019-06-25 | 卢松 | The zymotechnique of low-cost high-quality xanthan gum |
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