CN104372048A - Method for increasing yield of bacterial cellulose produced by fermenting acetobacter xylinum - Google Patents
Method for increasing yield of bacterial cellulose produced by fermenting acetobacter xylinum Download PDFInfo
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- CN104372048A CN104372048A CN201410726150.0A CN201410726150A CN104372048A CN 104372048 A CN104372048 A CN 104372048A CN 201410726150 A CN201410726150 A CN 201410726150A CN 104372048 A CN104372048 A CN 104372048A
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Abstract
The invention discloses an optimal compound recipe for producing bacterial cellulose by adding coconut juice, corn steep liquor, niacin, biotin, Twenn-80 and CMC into a basal culture medium to ferment acetobacter xylinum, and discloses a method for increasing yield of bacterial cellulose produced by fermenting acetobacter xylinum. The method includes steps of compounding substances of the same synergia mechanism and different synergia mechanisms to increase the yield of bacterial cellulose on the basis of determining the synergia mechanisms of the substances, adding 10%VOL of corn steep liquor, 1.0mg/L of niacin, 25mg/L of biotin and 4.0g/L of CMC into the basal culture medium, and statically culturing 8 d in a constant-temperature incubator at 30 DEG C. By the method, the yield of the bacterial cellulose is up to 9.326g/L and is 6.2 times of 1.506g/L in the basal culture medium without synergia substances, and the above compound recipe is the optimal combination for compounding.
Description
Technical field
The present invention relates to bacteria cellulose production field, specifically a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output.
Background technology
The present invention is various in style based on the synergistic agent of the existing element yielded poorly of current acetobacter xylinum fermented-producing bacteria cellulose, it is reported that tea, Sucus Cocois, corn steep liquor, cellulase, para-amino benzoic acid, alcohol, acetic acid, citric acid, lactic acid, niacinamide, Twenn-80, poly-interior alkene acid amides, vinylformic acid, poly-polysaccharide, agar, carboxymethyl cellulose (CMC), Microcrystalline Cellulose and sodium alginate etc. all can improve bacteria cellulose output.Due to different strains, substratum composition but in the mechanism of action of synergistic agent, opinions vary, thinking of having is beneficial to thalli growth, as nutritive substance etc. abundant in the various trace elements in para-amino benzoic acid, niacinamide, black tea water and VITAMIN, Sucus Cocois and corn steep liquor; Also the effect of black tea water is that inhibition of caffeine PDE activity wherein makes cyclic GMP increase and the synergism of generation to have research to think, the synergism of Sucus Cocois and corn steep liquor is the impact of wherein growth hormone, also have research think add corn steep liquor time, the cushion effect utilizing it strong can make fermented liquid maintain in stable pH value range, thus improves the output of bacteria cellulose; Some additives are unfavorable for thalli growth, but can increase the permeability of somatic cells film, as tensio-active agent Twenn-80 can change membrane passage, are conducive to cellular uptake nutrition and releasing product thus improve the output of bacteria cellulose; At synergistic agent in the Study on mechanism of acetobacter xylinum synthetic cellulose, both varied, there is again mutually contradictory or inconsistent place, and be all theory speculates, have not been reported and it is showed in the mode of some quantification of targets.Because different strains, substratum composition affect comparatively large on acetobacter xylinum fermented bacterial cellulose, be difficult to compare between the conclusion of report.Invention person utilizes same bacterial strain, same foundation substratum, and the Sucus Cocois of to its Synergistic Mechanisms having different versions, corn steep liquor, caffeine, Twenn-80, carboxyl methyl cellulose, sodium alginate, nicotinic acid, vitamin H eight kind additives relatively many to research are tested, test respective synergistic effect, on the basis of clear and definite Synergistic Mechanisms, carry out composite, to obtaining the highest compounded combination of bacteria cellulose output to identical Synergistic Mechanisms material and different Synergistic Mechanisms material respectively.
Summary of the invention
The invention provides a kind of method utilizing the composite raising bacteria cellulose output of synergistic matter.
The object of the invention is to solve in current a kind of novel biomedical material bacteria cellulose fermentation production process, due to the restriction of fermentation raw material and training method, cause the fermentation period of bacteria cellulose grow, yield poorly, the problem such as production cost height.China is mostly that fermention medium produces bacteria cellulose with Sucus Cocois, but the place of production of Sucus Cocois is limited to, output is less, production cost is high, the Chinese scholars output adopting the mode of adding synergistic matter to improve bacteria cellulose more, often study for one or more single synergistic matters, lack the compounded combination research of multiple synergistic matter, due to the difference of bacterial strain and culture medium, each result of study is made to lack comparability, simultaneously due to indefinite to Synergistic Mechanisms, the selection of synergistic matter is caused not have concrete theoretical foundation.This patent is for same bacterial strain, same foundation substratum, by eight kinds of synergistic matters that synergism class test is common, to the material having clear and definite synergism, with its Synergistic Mechanisms of quantifiable Study of indices, carry out composite to the synergistic matter of identical Synergistic Mechanisms and different Synergistic Mechanisms respectively, reach the object improving bacteria cellulose output.
technical scheme:
the research of single synergistic agent: in numerous synergistic agent to acetobacter xylinum fermented-producing bacteria cellulose, Selecting research relatively many and the Sucus Cocois (representative of suitability for industrialized production raw material) of having different versions to its Synergistic Mechanisms, corn steep liquor, caffeine, Twenn-80, CMC, sodium alginate, nicotinic acid, vitamin H eight kinds of additives add basic medium to, respectively by inoculation acetobacter xylinum fermentation synthesis bacteria cellulose.Found that: Sucus Cocois, corn steep liquor, Twenn-80, CMC, nicotinic acid and vitamin H six kinds of materials have promoter action for acetobacter xylinum synthesis bacterial fibers, and sodium alginate and caffeine have certain restraining effect.To having the Sucus Cocois of synergy, corn steep liquor, Twenn-80, CMC, nicotinic acid and vitamin H, testing out each synergistic agent the suitableeest addition added in basic medium and being respectively Sucus Cocois 50%VOL, corn steep liquor 10%VOL, Twenn-80 0.8g/L, CMC 4g/L, nicotinic acid 1.0mg/L, vitamin H 25mg/L.The acetobacter xylinum of different synergistic agent under the suitableeest addition produces bacteria cellulose output as shown in Figure 1, the synergistic effect of various synergistic agent is by weak order being by force: corn steep liquor > Sucus Cocois > vitamin H > nicotinic acid >Twenn-80>CMC> is blank, and present patent application contriver is the mechanism of action of each synergistic agent by a large amount of research and inquirement also.Sucus Cocois, corn steep liquor, nicotinic acid and vitamin H are beneficial to the growth of thalline; Corn steep liquor also increases the cushion effect of fermented liquid simultaneously; CMC changes the polymerization methods of bacteria cellulose; Twenn-80 adds membrane passage.The synergism (for blank 5.68 times) of corn steep liquor to acetobacter xylinum synthesis bacteria cellulose BC is better than BC industrial production and commonly uses raw material Sucus Cocois (3.78 times for blank), and corn steep liquor is Starch Production tankage, the suitability for industrialized production using it for BC can reduce Financial cost.
the research that synergistic agent is composite:in order to improve the output of acetobacter xylinum fermented bacterial cellulose further, the present inventor has carried out the discussion of Synergistic Mechanisms to the single synergistic agent that early stage has been determined, composite test is carried out to the synergistic matter with identical Synergistic Mechanisms, result is as Fig. 2, then carry out composite test to the synergistic matter with different Synergistic Mechanisms, result is as Fig. 3.It is best that test-results finds that the synergistic matter of different Synergistic Mechanisms carries out composite effect.
A kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output of the present invention, it is characterized in that synergistic matter corn steep liquor and one of carboxyl methyl cellulose, nicotinic acid, vitamin H, two or the composite synergistic matter of three kind of material, to join in the substratum of acetobacter xylinum to produce bacteria cellulose, reach volume increase object.
The formulation ratio of compound synergic material is:
(1) corn steep liquor+CMC+ nicotinic acid
(2) corn steep liquor+CMC+ vitamin H
(3) corn steep liquor+CMC+ nicotinic acid+vitamin H
(4) corn steep liquor+nicotinic acid
(5) corn steep liquor+vitamin H
(6) corn steep liquor+nicotinic acid+vitamin H
In each formula: often liter of basic medium aqueous solution containing acetobacter xylinum is 0.1 L containing corn steep liquor, is 0.9 ~ 1.1mg containing nicotinic acid, is 23 ~ 27 mg containing vitamin H, is 3.5 ~ 4.5g containing CMC.
Indication basic medium proportioning is that often liter of basic medium aqueous solution is containing sucrose 45 ~ 55g, yeast extract paste 8 ~ 12g, peptone 6 ~ 10g, potassium primary phosphate 4.1 ~ 4.5g, magnesium sulfate 2.2 ~ 2.7g.
Optimum proportioning, the formula of above-mentioned compound synergic material for (3) are: corn steep liquor+CMC+ nicotinic acid+vitamin H, the nutrient solution containing acetobacter xylinum by often liter adds corn steep liquor 0.1 L, nicotinic acid 1.0mg, vitamin H 25 mg and CMC 4.0g.
Adding compound synergic material optimum proportioning formula is (3), improve 6.2 times under the same conditions than the basic medium bacteria cellulose output not adding synergistic matter, the bacteria cellulose output of addition manner more single or composite than other any synergistic matters all increases under the same conditions.
The processing step that the present invention adds compound synergic material production bacteria cellulose is:
(1) by above-mentioned formula, proportioning, basic medium is prepared with distilled water;
(2) by above-mentioned formula, proportioning, synergistic matter is added in basic medium respectively;
(3) by the basic medium of step (2) gained in 121 DEG C of sterilizing 20 ~ 30min, be chilled to room temperature;
(4) the liquid acetobacter xylinum seed liquor of culture volume 8% is accessed;
Quiescent culture 8d in (5) 30 DEG C of constant incubators;
(6) collect bacteria cellulose film, repeatedly rinse except after the substratum on striping surface and impurity with distilled water, be placed in 80 DEG C, in the NaOH solution of 0.1mol/L, maintain 2h, to remove tropina and residual media, be creamy white to film translucent, be cooled to room temperature;
(7) by the bacteria cellulose film of step (6) with in 0.1mol/L HCl and 30min, distilled water fully washs;
(8) step (7) is dried to constant weight at 80 DEG C and namely obtains product.
The liquid acetobacter xylinum seed liquor of above-mentioned inoculation is the nutrient solution of acetobacter xylinum inclined-plane kind to 50mL sterilizing of inoculation 3 ring activation, 28 DEG C, 100r/min shaking table cultivation 24h.
beneficial effect:
The present inventor will determine that the synergistic matter of Synergistic Mechanisms carries out composite according to identical Synergistic Mechanisms and different Synergistic Mechanisms, be a fermentation period with 8 days, show that best compounded combination is corn steep liquor 10%VOL, nicotinic acid 1.0mg/L, vitamin H 25mg/L and CMC 4.0g/L, when it being added, the output of bacteria cellulose is up to 9.326g/L simultaneously, is 6.2 times of the blank group bacteria cellulose output 1.506g/L not adding any synergistic matter.
The invention provides a kind of on the basis of clear and definite Synergistic Mechanisms by method composite for synergistic matter, the object improving bacteria cellulose output can be reached, solve the difficult problem that basic medium fermented-producing bacteria cellulose yields poorly, add the output of bacteria cellulose, reduce production cost, improve economic benefit.It is China's the first that the basis of clear and definite Synergistic Mechanisms is carried out composite to synergistic matter.
Accompanying drawing explanation
The synergistic effect of the single synergistic matter of Fig. 1 compares
The fine plain output of bacterium that Fig. 2 same mechanism synergistic matter is composite
The bacteria cellulose output that the different mechanism synergistic matter of Fig. 3 is composite
In figure, each synergistic matter addition is as the criterion to prepare 1 liter of substratum: Sucus Cocois 0.5L, corn steep liquor 0.1 L, nicotinic acid 1.0mg, vitamin H 25 mg, Twenn-80 0.8g and CMC4.0g.
embodiment
embodiment 1single synergistic agent improves the test of acetobacter xylinum fermented bacterial cellulose output.
Step: the preparation of (1) substratum: take sucrose 50g, yeast extract paste 10g, peptone 8g, potassium primary phosphate 4.3g, magnesium sulfate 2.5g, dissolve with distilled water respectively, to add in the volumetric flask of 1 liter and constant volume, prepare 7 parts altogether, be numbered No1, No2, No3, No4, No5, No6, No7. respectively
(2) in No1, add 500mL Sucus Cocois, in No2, add 100mL corn steep liquor, add 0.8 g Twenn-80 in No3, in No4, add 4g CMC, in No5, add 1.0 mg nicotinic acid, add in No6 in 25 mg vitamin Hs, No7 and do not add any synergistic agent, do blank.
(3) by the basic medium of step (2) gained in 121 DEG C of sterilizing 20 ~ 30min, be chilled to room temperature;
(4) in No1 ~ No7, access 80mL liquid acetobacter xylinum seed liquor;
(5) by the No1 ~ No7 of step (4) quiescent culture 8d in 30 DEG C of constant incubators;
(6) collect bacteria cellulose film, repeatedly rinse except after the substratum on striping surface and impurity with distilled water, be placed in 80 DEG C, in the NaOH solution of 0.1mol/L, maintain 2h, to remove tropina and residual media, be creamy white to film translucent, be cooled to room temperature;
(7) by the bacteria cellulose film of step (6) with in 0.1mol/L HCl and 30min, distilled water fully washs;
(8) step (7) is dried to constant weight at 80 DEG C and namely obtains product.
The liquid acetobacter xylinum seed liquor of inoculation is the nutrient solution of acetobacter xylinum inclined-plane kind to 50mL sterilizing of inoculation 3 ring activation, 28 DEG C, 100r/min shaking table cultivation 24h.
Single synergistic agent improves the result of acetobacter xylinum fermented bacterial cellulose output as Fig. 1, and as can be seen from Figure 1, corn steep liquor effect of increasing production is better than Sucus Cocois, and cost is lower.
embodiment 2the test of same mechanism synergistic matter composite raising acetobacter xylinum fermented bacterial cellulose output.
Step (1), with embodiment 1 step (1), prepares 11 parts, numbering 1 ~ 11 altogether.
(2) in numbering 1, Sucus Cocois 500mL is added, corn steep liquor 100mL, Sucus Cocois 500mL is added in numbering 2, nicotinic acid 1.0 mg, Sucus Cocois 500mL is added in numbering 3, vitamin H 25 mg, Sucus Cocois 500mL is added in numbering 4, corn steep liquor 100mL, nicotinic acid 1.0 mg, Sucus Cocois 500mL is added in numbering 5, corn steep liquor 100mL, vitamin H 25 mg, Sucus Cocois 500mL is added in numbering 6, nicotinic acid 1.0 mg, vitamin H 25 mg, Sucus Cocois 500mL is added in numbering 7, corn steep liquor 100mL, nicotinic acid 1.0 mg, vitamin H 25 mg, corn steep liquor 100mL is added in numbering 8, nicotinic acid 1.0 mg, corn steep liquor 100mL is added in numbering 9, vitamin H 25 mg, corn steep liquor 100mL is added in numbering 10, nicotinic acid 1.0 mg, vitamin H 25 mg, nicotinic acid 1.0 mg is added in numbering 11, vitamin H 25 mg.
(3) with (3) of embodiment 1.
(4) in numbering 1 ~ 11, access 80mL liquid acetobacter xylinum seed liquor;
(5) by the numbering 1 ~ 11 of step (4) quiescent culture 8d in 30 DEG C of constant incubators;
(6) (7) (8) are with (6) (7) (8) of embodiment 1.
The liquid acetobacter xylinum seed liquor of inoculation is with embodiment 1.
Identical Synergistic Mechanisms synergistic matter composite raising acetobacter xylinum fermented bacterial cellulose output, test-results is as Fig. 2, and as can be seen from Figure 2, numbering 8,9,10 effect of increasing production is best.
embodiment 3the test of different mechanism synergistic matters composite raising acetobacter xylinum fermented bacterial cellulose output.
Step (1), with embodiment 1 step (1), is prepared 10 parts altogether, is numbered 1# ~ 10# respectively.
(2) in 1#, corn steep liquor 100mL is added, nicotinic acid 1.0 mg, Twenn-80 0.8 g, corn steep liquor 100mL is added in 2#, vitamin H 25 mg, Twenn-80 0.8 g, corn steep liquor 100mL is added in 3#, nicotinic acid 1.0 mg, Twenn-80 0.8 g, vitamin H 25 mg, corn steep liquor 100mL is added in 4#, nicotinic acid 1.0 mg, CMC 4 g, corn steep liquor 100mL is added in 5#, vitamin H 25 mg, CMC 4 g, corn steep liquor 100mL is added in 6#, nicotinic acid 1.0 mg, vitamin H 25 mg, CMC 4 g, corn steep liquor 100mL is added in 7#, nicotinic acid 1.0 mg, Twenn-80 0.8 g, CMC 4 g, corn steep liquor 100mL is added in 8#, vitamin H 25 mg, Twenn-80 0.8 g, CMC 4 g, corn steep liquor 100mL is added in 9#, nicotinic acid 1.0 mg, vitamin H 25 mg, Twenn-80 0.8 g, CMC 4 g, Twenn-80 0.8 g is added in 10#, CMC 4 g.
(3) with embodiment 1(3).
(4) in numbering 1# ~ 10#, access 80mL liquid acetobacter xylinum seed liquor;
(5) by the numbering 1# of step (4) ~ 10# quiescent culture 8d in 30 DEG C of constant incubators;
(6) (7) (8) are with (6) (7) (8) of embodiment 1.
The liquid acetobacter xylinum seed liquor of inoculation is with embodiment 1.
Different Synergistic Mechanisms synergistic matters composite raising acetobacter xylinum fermented bacterial cellulose output, test-results is as Fig. 3, and as can be seen from Figure 3, numbering 4#, 5#, 6# effect of increasing production are outstanding, wherein it is preferred that 6#.
In above-described embodiment, each sample of often kind of scheme at least does 5 Duplicate Samples, takes average result.
Claims (7)
1. one kind is improved the method for acetobacter xylinum fermented-producing bacteria cellulose output, it is characterized in that synergistic matter corn steep liquor and one of carboxyl methyl cellulose, nicotinic acid, vitamin H, two or the composite synergistic matter of three kind of material, to join in the substratum of acetobacter xylinum to produce bacteria cellulose, reach volume increase object.
2. a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output according to claim 1, is characterized in that the formulation ratio of compound synergic material is:
(1) corn steep liquor+CMC+ nicotinic acid
(2) corn steep liquor+CMC+ vitamin H
(3) corn steep liquor+CMC+ nicotinic acid+vitamin H
(4) corn steep liquor+nicotinic acid
(5) corn steep liquor+vitamin H
(6) corn steep liquor+nicotinic acid+vitamin H
Each formula kind: often liter of basic medium aqueous solution containing acetobacter xylinum is 0.1 L containing corn steep liquor, is 0.9 ~ 1.1mg containing nicotinic acid, is 23 ~ 27 mg containing vitamin H, is 3.5 ~ 4.5g containing CMC.
3. a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output according to claim 1 and 2, is characterized in that indication basic medium proportioning is for often liter of basic medium aqueous solution is containing sucrose 45 ~ 55g, yeast extract paste 8 ~ 12g, peptone 6 ~ 10g, potassium primary phosphate 4.1 ~ 4.5g, magnesium sulfate 2.2 ~ 2.7g.
4. a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output according to claim 2, it is characterized in that the optimum proportioning of compound synergic material, formula is: (3) corn steep liquor+CMC+ nicotinic acid+vitamin H, add corn steep liquor 0.1 L, nicotinic acid 1.0mg, vitamin H 25 mg and CMC 4.0g by often liter of nutrient solution containing acetobacter xylinum.
5. a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output according to claim 4, it is characterized in that adding compound synergic material improves 6.2 times under the same conditions than the basic medium bacteria cellulose output not adding synergistic matter, the bacteria cellulose output of addition manner more single or composite than other any synergistic matters all increases under the same conditions.
6. a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output according to claim 1, is characterized in that the processing step adding compound synergic material production bacteria cellulose is:
(1) by formula, proportioning, basic medium is prepared with distilled water;
(2) by formula, proportioning, synergistic matter is added in basic medium respectively;
(3) by the basic medium of step (2) gained in 121 DEG C of sterilizing 20 ~ 30min, be chilled to room temperature;
(4) the liquid acetobacter xylinum seed liquor of culture volume 8% is accessed;
Quiescent culture 8d in (5) 30 DEG C of constant incubators;
(6) collect bacteria cellulose film, repeatedly rinse except after the substratum on striping surface and impurity with distilled water, be placed in 80 DEG C, in the NaOH solution of 0.1mol/L, maintain 2h, to remove tropina and residual media, be creamy white to film translucent, be cooled to room temperature;
(7) by the bacteria cellulose film of step (6) with in 0.1mol/L HCl and 30min, distilled water fully washs;
(8) step (7) is dried to constant weight at 80 DEG C and namely obtains product.
7. a kind of method improving acetobacter xylinum fermented-producing bacteria cellulose output according to claim 6, it is characterized in that the liquid acetobacter xylinum seed liquor inoculated is the nutrient solution of acetobacter xylinum inclined-plane kind to 50mL sterilizing of inoculation 3 ring activation, 28 DEG C, 100r/min shaking table cultivation 24h.
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CN105368896A (en) * | 2015-12-16 | 2016-03-02 | 贵州大学 | Method for optimizing inulin fermentation medium to increase yield of dry bacterial cellulose films and improve quality of dry bacterial cellulose films |
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CN106047960A (en) * | 2016-07-22 | 2016-10-26 | 南京理工大学 | Optimizing method for ferment-producing bacterial cellulose by using gluconic acid as sole carbon source |
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CN113881733A (en) * | 2021-11-23 | 2022-01-04 | 广东药科大学 | Method for preparing food gum-based bacterial cellulose by using acid hydrolysis liquid of rice processing byproduct |
CN113881733B (en) * | 2021-11-23 | 2024-04-05 | 广东药科大学 | Method for preparing food gum base bacterial cellulose by using rice processing byproduct acid hydrolysate |
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