CN102337309A - Potato residue culture medium - Google Patents
Potato residue culture medium Download PDFInfo
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- CN102337309A CN102337309A CN2011102986793A CN201110298679A CN102337309A CN 102337309 A CN102337309 A CN 102337309A CN 2011102986793 A CN2011102986793 A CN 2011102986793A CN 201110298679 A CN201110298679 A CN 201110298679A CN 102337309 A CN102337309 A CN 102337309A
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- potato
- nutrient solution
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- waste residue
- glycerine
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Abstract
The invention relates to a bacterial cellulose culture medium, which is a potato culture fluid. The technical scheme of the potato culture fluid provided by the invention specifically comprises the steps of: taking potato residues, adding water, and heating at a temperature of 80-100 DEG C for not less than 1 hour to make the mass ratio of potato to distilled water 2-11:50, so as to obtain the potato culture fluid, wherein the pH value of the potato culture fluid is 4.0-7.5; and mixing glycerin and the potato culture fluid to obtain a culture fluid, wherein the mass ratio of glycerin to potato in the potato culture fluid is 2-13:20-110. According to the invention, the potato residues after potato starch production are firstly used as a raw material for a culture medium and are treated by using the simplest method to obtain the culture medium for culturing bacterial cellulose; the culture medium has low cost and high yield rate, the problem of recycling the potato residues is simultaneously solved, and the culture time is short; in addition, the invention provides an important condition for stable and large-scale bacterial cellulose culture.
Description
Technical field
The present invention relates to microorganism field, particularly the substratum of refuse reclamation preparation.
Background technology
Potato is that a kind of distribution is extensive, and flexibility is strong, the easy raise crop that output is high, and except as the grain, potato is mainly used in starch processing.Utilize one ton of starch of the every production of potato will produce 7.5 tons of potato waste residues.Mainly containing protein, amino acid and a small amount of glucide in the potato waste residue, is the resource of recyclable utilization.At present, the potato waste residue is recyclable is used to produce alcohol, feed, degradable plastics and extraction pectin and prepares food fibre etc.But in recycling, the potato waste residue usually faces following problem: 1) potato waste residue viscosity is high, is difficult for solid-liquid separation; 2) time-consuming expensive before the recovery to the sterilization of potato waste residue.
Bacteria cellulose is the another kind of Mierocrystalline cellulose except that plant cellulose; Be the natural inert material of bacterium synthetic; Bacteria cellulose (bacterial cellulose; BC) having excellent bioaffinity, biocompatibility, biological fitness and good biodegradability, is that the new bio of the excellent performance of generally acknowledging is in the world learned material.Application mainly concentrates on the bio-medical material of high added value to BC at present, like aspects such as tissue engineering bracket, artificial blood vessel and artificial skins.The bacterium that can produce with bacteria cellulose mainly contains
Acetobacter,
Rhizobium, AgrobacteriumWith
SarcinaDeng, what wherein output was the highest is acetobacter xylinum,
Acetobacter xylinus, belong to Gram-negative (minority is variable); Strict aerobic, respiratory metabolism is from nonfermented; Traditional view is thought: ethanol, glycerine and lactic acid are best carbon sources.Though acetobacter xylinum output is higher, but still can not solve the high problem of bacteria cellulose production cost.The method that adds the culture medium culturing bacteria cellulose with glycerine is disclosed in " research of the synthetic bacteria cellulose of Acetobacter pasteurianus " literary composition; Filter out through orthogonal test: the Acetobacter pasteurianus inoculum size with 7% is cultivated under 31 ℃ of conditions; Main carbon source is that massfraction is 3% glycerine, and the production peak of its bacteria cellulose is 2.326%.But, adopted higher Carnis Bovis seu Bubali cream of price and peptone staple in this method, and only carried out the small-scale cultivation of 10mL system as substratum.As everyone knows, in field of microorganism engineering, from small-scale experiment screening to the industrialization production more than the pilot scale; The condition of each link all must be optimized; Wherein, also comprise Optimum of culture medium, as without optimization; The lab scale experiment condition is mechanically enlarged in large-scale production, will face pH value, temperature and cultivate can't implementing that desired gas controllability problem causes.In addition, the technology barrier of bacteria cellulose is that mainly fermentation level is lower, yields poorly, cost is high, price is not supported common plant cellulose; The 2nd, further study and utilize the film forming of bacteria cellulose and the also not solution of Technology of moulding.How improving cellulose output, reduce cost, further improve its output thereby seek more cheap better bacteria cellulose raw materials for production, will be the basis of bacteria cellulose research.
Summary of the invention
One of the object of the invention is to provide a kind of potato nutrient solution, and this potato nutrient solution can serve as the non-main carbon source medium component that acetobacter xylinum prepares bacteria cellulose, and this substratum is applicable to the scale operation of bacteria cellulose.
For realizing above-mentioned purpose, technical scheme of the present invention is:
Potato nutrient solution, the beans that fetch earth add water heats under 80-100 ℃ of condition and is not less than 1 hour, and the mass ratio that makes potato and zero(ppm) water is 2-11:50, regulate the pH value to the 4.0-7.5 the potato nutrient solution.
Further, said potato is the remaining part of preparation potato starch, i.e. potato waste residue;
Further, the pH value of said potato nutrient solution is 6.8.
Two of the object of the invention is to provide a kind of substratum of bacteria cellulose, and this culture medium cost is cheap, is applicable to large-scale preparation bacteria cellulose.
For realizing above-mentioned purpose, technical scheme of the present invention is:
The substratum that contains described potato nutrient solution contains potato nutrient solution and glycerine in the said substratum, the mass ratio of the potato in said glycerine and the potato nutrient solution is 2-13:20-110.
Further, said substratum is by potato nutrient solution, glycerine, K
2HPO
4Form potato in the said potato nutrient solution and glycerine, K with yeast extract paste
2HPO
4With the mass ratio of yeast extract paste be 40-220:4-26:1-5:3-14, and pH is transferred to 5.0-7.5, the said substratum that contains the potato nutrient solution.
Further, said substratum is by potato nutrient solution, glycerine, K
2HPO
4Form potato waste residue in the said potato nutrient solution and glycerine, K with yeast extract paste
2HPO
4With the mass ratio of yeast extract paste be 40-220:8:3:8, and pH is transferred to 6.8, the said substratum that contains the potato nutrient solution.
Beneficial effect of the present invention is: the present invention will produce starch potato waste residue afterwards first as culture medium raw material; After the most simply method is handled the potato waste residue, obtain being used for culturing bacterium Mierocrystalline cellulose substratum; This culture medium cost is cheap; It is high to be used for culturing bacterium Mierocrystalline cellulose yield, and what the present invention had solved the potato waste residue simultaneously utilizes problem again.Potato waste residue nutrient solution and glycerine are share in the culturing bacterium Mierocrystalline cellulose at certain proportioning second line of a couplet, and its productive rate is high, and cost is low, and can shorten incubation time.In addition, the present invention cultivates for the stable extensive bacteria cellulose of bacteria cellulose essential condition is provided.
Embodiment
Acetobacter xylinum among the following embodiment (gluconacetobacter xylium), available from China Committee for Culture Collection of Microorganisms common micro-organisms center, bacterium numbering is 1.2378.
Following bacteria cellulose freezing dry process: be that bacteria cellulose is tiled in the lyophilize dish, carry out from-30 ℃---25 ℃ of temperature control vacuum freezedryings step by step are until the bacteria cellulose complete drying.It is solid-state that freeze-drying utilizes the deep-frozen method that the interior water of bacterial cellulose wet-coating is become, and in vacuum freeze drier, utilizes the method for its vacuum tightness of control then, makes the water sublimate of film internal solid.And, cause material to dehydrate with the steam that condensation method is caught and condensation distils.The vacuum lyophilization process is carried out under the condition of extremely low temperature and high vacuum, and is little to the thermally denature of material, on to greatest extent, kept the BA of material.Simultaneously, in the process of distillation, can not destroy its physical structure, changes of chemical structures is also very little, therefore can preserve the tridimensional network of bacteria cellulose film well.
The preparation of embodiment 1 potato waste residue
The waste residue of potato described in the present invention is the residuum after the bright article of potato extract starch.Concrete preparation method is: the beans that fetch earth, clean the surperficial earth of potato, and remove foreign matters such as stone and iron filings.With kibbler the mealy potato of cleaning is broken into paste to twisting with the fingers no granular sensation with hand.An amount of injection clear water quickens flowing of the potato dregs of rice, and starch is better separated with the potato dregs of rice.It is 130 times/minute that the potato dregs of rice are used vibration number, and the plansifter that sieve aperture is 30 orders, 50 orders, 60 orders from front to back, 80 orders are gradually little sieves, and starch flows into sieve down, and the potato waste residue is from sieving outflow, and its humidity is 75%.Also can adopt additive method to separate the starch in the potato, and then obtain the potato waste residue.
Go the potato waste residue under 100 ℃ of (80 ℃ also can) conditions, to heat 1 hour, potato waste residue nutrient solution.
Embodiment 2 potato waste residue inoculum preparation bacteria celluloses
The preparation of 1 bacteria cellulose
The cultivation of in substratum, recovering of acetobacter xylinum bacterial classification inoculation, dynamically shaking table is cultivated, and 30 ℃, 2-3 days; BC fermentation culture with dynamically increasing behind the bacterium is transferred in the tray, 30 ℃, static cultivation 14 days, BC, for membranaceous.To cultivate the BC that obtains and put into 90 ° of C zero(ppm) water constant temperature 2 hours, and take out the back and clean 2 times with zero(ppm) water; NaOH solution with ebullient 0.5 mol/L boiled 15 minutes; Putting into massfraction and be 1% NaOH solution left standstill 2 days; Be placed on and be dipped to neutrality in the deionized water, get BC, be the BC wet film; Said wet film lyophilize is preserved, get BC, be the BC dry film.
2 potato nutrient solutions
The beans waste residue that fetches earth adds water (80 ℃ also can) under 100 ℃ of conditions heated 1 hour, the potato nutrient solution, the mass ratio of potato waste residue and zero(ppm) water (potato waste residue concentration), potential of hydrogen and productive rate see table 1 for details.
Table 1 potato raw slag material prepares bacteria cellulose film
| Potato waste residue concentration (%) | PH | Cellulose membrane weight in wet base mean (g/L) | Cellulose membrane dry weight mean (g/L) |
| 4 | 5.0 | 28.5 | 3.0 |
| 4 | 6.8 | 29.45 | 3.1 |
| 22 | 6.8 | 31.6 | 3.3 |
| 22 | 7.5 | 28.5 | 3.0 |
What the potato waste residue was traditional utilizes mode for producing alcohol, feed, degradable plastics and extracting pectin and the preparation food fibre again.But in the process of above-mentioned recycling, or it is high to relate to potato waste residue viscosity, not easily separated, or not only time-consuming but also expensive sterilizing works before relating to the potato waste residue and reclaiming, and makes industrialization utilize the project of potato waste residue and few.
Present embodiment is that the utilization again of potato waste residue provides new approaches, can the potato waste residue directly be used as culture medium raw material under the situation of not adding other raw materials.Though use the potato nutrient solution of potato waste residue preparation separately, the productive rate of its bacteria cellulose is not high,, present embodiment has confirmed that first the potato waste residue can serve as the bacteria cellulose medium component.Present embodiment provides new solution for producing the high culture medium cost of bacteria cellulose.The potato nutrient solution of potato waste residue preparation mixes with other nutritive ingredients, can prepare the bacteria cellulose special culture media.
In addition; From in logic can be beyond all doubt draw: the potato waste residue is that the potato nutrient solution of raw material possesses under the prerequisite of the ability of serving as acetobacter xylinum inoculum preparation bacteria cellulose, and potato also possesses the ability of serving as acetobacter xylinum inoculum preparation bacteria cellulose.
Embodiment 3 potato waste residue glycerin mediums
In embodiment 2, adopt single potato waste residue to make culture medium raw material, the productive rate of its bacteria cellulose is 3.0-3.3%; The potato waste residue is that most of starch in the potato is extracted the last residue in back, and the growth of bacteria cellulose is to need an amount of carbon source and nitrogenous source, so, if can in the potato waste residue, replenish an amount of carbon source, then can improve the output of bacteria cellulose.When carrying out the screening of carbon source, must satisfy three conditions: the one, cheap, otherwise just lost the advantage cheaply of refuse reclamation; The 2nd, must be able to adapt to the scale operation of industrialization, through laboratory scale amplification culture, still can keep the stable of flora; The 3rd, productive rate is high, if the productive rate of uniting utilization (by production cost) of potato waste residue and other carbon sources is less than or equal to the effect of potato starch itself, then its productive rate of illustrative is not high.
The preparation of 1 bacteria cellulose
The cultivation of in substratum, recovering of acetobacter xylinum bacterial classification inoculation, dynamically shaking table is cultivated, and 30 ℃, 3 days; BC fermentation culture with dynamically increasing behind the bacterium is transferred in the tray, 30 ℃, static cultivation 7 days, BC, for membranaceous.To cultivate the BC that obtains and put into 90 ° of C zero(ppm) water constant temperature 2 hours, and take out the back and clean 2 times with zero(ppm) water; NaOH solution with ebullient 0.5 mol/L boiled 15 minutes; Putting into massfraction and be 1% NaOH solution left standstill 2 days; Be placed on and be dipped to neutrality in the deionized water, get BC, be the BC wet film; Said wet film lyophilize is handled and preserved, get BC, be the BC dry film.The preparation of bacteria cellulose is in the culture tank of 5L scale, to carry out.
The substratum of 2 potato nutrient solutions and glycerine mixing gained
The potato nutrient solution is the potato nutrient solution of feedstock production with the potato waste residue for embodiment 1 is described in the present embodiment.In the substratum of said potato nutrient solution and glycerine mixing gained, the mass ratio of potato waste residue and glycerine, substratum potential of hydrogen and bacteria cellulose productive rate see table 2 for details.
The potato raw slag material that table 2 adds glycerine prepares bacteria cellulose film
| Potato waste residue and qualities of glycerin ratio | PH | Cellulose membrane weight in wet base mean (g/L) | Cellulose membrane dry weight mean (g/L) |
| 55:1 | 5.0 | 34.2 | 3.6 |
| 110:13 | 6.8 | 37.05 | 3.9 |
| 10:1 | 6.8 | 33.25 | 3.5 |
| 20:13 | 7.5 | 33.3 | 3.5 |
Disclose with glycerine as the cellulosic method of carbon source culturing bacterium in " research of the synthetic bacteria cellulose of Acetobacter pasteurianus " literary composition; They are through screening the proportioning of glycerine consumption; Think when massfraction is 3% consumption; The nitrogenous source higher with other prices (like peptone, Carnis Bovis seu Bubali cream etc.) united utilization, under the petridish scale, cultivates, and its productive rate is 2.326g/L.
In the present embodiment, the consumption that the glycerine consumption of 0.4-2.6% has overcome the glycerine of " research of the synthetic bacteria cellulose of Acetobacter pasteurianus " proposition is the 3% o'clock the highest technological prejudice of output at massfraction.With respect to the cost of potato waste residue, the glycerine price is more expensive relatively, and in " research of the synthetic bacteria cellulose of Acetobacter pasteurianus " literary composition, present embodiment only then can minimumly reduce production costs 9 one-tenth on carbon source.And, in the present embodiment, be in the culture tank of 5L, to carry out, on scale, the cultivation scale of present embodiment is obviously greater than " research of the synthetic bacteria cellulose of an Acetobacter pasteurianus " literary composition.In addition, potato waste residue and glycerine combined utilization make, and its incubation time has shortened half the.
Embodiment 4 full substratum
The preparation of 1 bacteria cellulose
The cultivation of in substratum, recovering of acetobacter xylinum bacterial classification inoculation, dynamically shaking table is cultivated, and 30 ℃, 2 days; BC fermentation culture with dynamically increasing behind the bacterium is transferred in the tray, 30 ℃, static cultivation 7 days, BC, for membranaceous.To cultivate the BC that obtains and put into 90 ° of C zero(ppm) water constant temperature 2 hours, and take out the back and clean 2 times with zero(ppm) water; NaOH solution with ebullient 0.5 mol/L boiled 15 minutes; Putting into massfraction and be 1% NaOH solution left standstill 2 days; Be placed on and be dipped to neutrality in the deionized water, get BC, be the BC wet film; Said wet film lyophilize is preserved, get BC, be the BC dry film.The preparation of bacteria cellulose is in the culture tank of 5L scale, to carry out.
2 culture medium preparation
Prescription one: the potato nutrient solution 220g/L that the potato waste residue is prepared from, glycerine 26g/L, K
2HPO
45g, yeast extract paste 14g/L, acetic acid is transferred pH value to 7.5.
Prescription two: the potato nutrient solution 220g/L that the potato waste residue is prepared from, glycerine 26g/L, K
2HPO
45g, yeast extract paste 14g/L, acetic acid is transferred pH value to 5.0.
Prescription three: the potato nutrient solution 40g/L that the potato waste residue is prepared from, glycerine 4g/L, K
2HPO
41g, yeast extract paste 3g/L, acetic acid is transferred pH value to 7.5.
Prescription four: the potato nutrient solution 40g/L that the potato waste residue is prepared from, glycerine 4g/L, K
2HPO
41g, yeast extract paste 3g/L, acetic acid is transferred pH value to 5.0.
Its productive rate sees table 3 for details
The potato raw slag material that table 3 adds glycerine prepares bacteria cellulose film
| Prescription | Incubation time (my god) | Cellulose membrane weight in wet base mean (g/L) | Cellulose membrane dry weight mean (g/L) |
| Prescription one | 7 | 35.1 | 3.9 |
| Prescription two | 8 | 36.9 | 4.1 |
| Prescription three | 14 | 37.8 | 4.2 |
| Prescription four | 12 | 36 | 4.0 |
Through the bacteria cellulose film of basal culture medium preparation, the price that warp calculating is every square centimeter is merely 0.13 yuan.
In the above-mentioned substratum, the inoculum size of acetobacter xylinum bacterial classification is 7%, promptly gets 10
67 milliliters of cfu/ml acetobacter xylinums are inoculated in 93 milliliters the substratum.
Explanation is at last; Above embodiment is only unrestricted in order to technical scheme of the present invention to be described; Although through invention has been described with reference to the preferred embodiments of the present invention; But those of ordinary skill in the art should be appreciated that and can make various changes to it in form with on the details, and the spirit and scope of the present invention that do not depart from appended claims and limited.
Claims (6)
1. potato nutrient solution is characterized in that: the beans that fetch earth add water heats under 80-100 ℃ of condition and is not less than 1 hour, and the mass ratio that makes potato and zero(ppm) water is 2-11:50, regulate the pH value to the 4.0-7.5 the potato nutrient solution.
2. potato nutrient solution according to claim 1 is characterized in that: said potato is the remainder behind the preparation potato starch, i.e. potato waste residue.
3. potato nutrient solution according to claim 2 is characterized in that: the pH value of said potato nutrient solution is 6.8.
4. contain the substratum of each described potato nutrient solution of claim 1-3, it is characterized in that: contain potato nutrient solution and glycerine in the said substratum, the mass ratio of the potato in said glycerine and the potato nutrient solution is 2-13:20-110.
5. substratum according to claim 4 is characterized in that said substratum is by potato nutrient solution, glycerine, K
2HPO
4Form potato in the said potato nutrient solution and glycerine, K with yeast extract paste
2HPO
4With the mass ratio of yeast extract paste be 40-220:4-26:1-5:3-14, and pH is transferred to 5.0-7.5, the said substratum that contains the potato nutrient solution.
6. substratum according to claim 5 is characterized in that said substratum is by potato nutrient solution, glycerine, K
2HPO
4Form potato waste residue in the said potato nutrient solution and glycerine, K with yeast extract paste
2HPO
4With the mass ratio of yeast extract paste be 40-220:8:3:8, and pH is transferred to 6.8, the said substratum that contains the potato nutrient solution.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110298679.3A CN102337309B (en) | 2011-09-28 | 2011-09-28 | Potato residue culture medium |
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|---|---|---|---|
| CN201110298679.3A CN102337309B (en) | 2011-09-28 | 2011-09-28 | Potato residue culture medium |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107502631A (en) * | 2017-09-28 | 2017-12-22 | 上海应用技术大学 | A kind of production method of candida utili β D glucans |
| CN108315371A (en) * | 2018-04-20 | 2018-07-24 | 北京理工大学珠海学院 | A kind of cultural method of bacteria cellulose |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1440637A (en) * | 2003-04-01 | 2003-09-10 | 刘永昶 | Liquid edible fungus strain producing method and apparatus |
| CN101633938A (en) * | 2009-09-01 | 2010-01-27 | 山西省生物研究所 | Preparation method of fuel alcohol by fermenting waste potato dregs |
-
2011
- 2011-09-28 CN CN201110298679.3A patent/CN102337309B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1440637A (en) * | 2003-04-01 | 2003-09-10 | 刘永昶 | Liquid edible fungus strain producing method and apparatus |
| CN101633938A (en) * | 2009-09-01 | 2010-01-27 | 山西省生物研究所 | Preparation method of fuel alcohol by fermenting waste potato dregs |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107502631A (en) * | 2017-09-28 | 2017-12-22 | 上海应用技术大学 | A kind of production method of candida utili β D glucans |
| CN108315371A (en) * | 2018-04-20 | 2018-07-24 | 北京理工大学珠海学院 | A kind of cultural method of bacteria cellulose |
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| CN102337309B (en) | 2014-06-11 |
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Effective date of registration: 20160804 Address after: The 100070 Beijing Seahawks Fengtai District Science City Road No. 9 Patentee after: BEIJING GUANLAN TECHNOLOGY CO., LTD. Address before: Shinan District 266071 Shandong city of Qingdao province Xuzhou Road 32, building 2, unit 2, 601 households Patentee before: Wu Like |