CN105368896A - Method for optimizing inulin fermentation medium to increase yield of dry bacterial cellulose films and improve quality of dry bacterial cellulose films - Google Patents
Method for optimizing inulin fermentation medium to increase yield of dry bacterial cellulose films and improve quality of dry bacterial cellulose films Download PDFInfo
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- CN105368896A CN105368896A CN201510942263.9A CN201510942263A CN105368896A CN 105368896 A CN105368896 A CN 105368896A CN 201510942263 A CN201510942263 A CN 201510942263A CN 105368896 A CN105368896 A CN 105368896A
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Abstract
The invention discloses a method for optimizing an inulin fermentation medium to increase the yield of dry bacterial cellulose films and improve the quality of the dry bacterial cellulose films. Main components in the inulin fermentation medium for fermenting bacterial cellulose are screened with a Plackett-Burman experiment design method, absolute ethyl alcohol with the remarkable effect serves as a key factor to be further optimized, and an optimized inulin fermentation medium formula ratio is obtained. The yield of the bacterial cellulose fermented through the optimized inulin fermentation medium is 5.54 g/L, is 21.31 times that (0.26 g/L) of the dry bacterial cellulose films fermented through an inulin medium base material, and is increased by 15.42% compared with the yield (4.80 g/L) of the dry bacterial cellulose films fermented through a basic medium, the main quality indexes are better than the main quality indexes of the bacterial cellulose cultured through the basic medium, and the bacterial cellulose prepared through the optimized inulin fermentation medium is better than that prepared through the basic medium in the performance aspects of the water binding capacity, the crystallinity, the heat stability and the like.
Description
Technical field
The present invention relates to bacteria cellulose production field, specifically, is a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality.
Background technology
Sucus Cocois is the earliest for the production of the raw material of bacteria cellulose, is also the raw material generally used at present.In recent years, bacteria cellulose relies on its excellent characteristic to be widely used in numerous areas such as medical treatment, weaving, environmental protection, papermaking, cause bacteria cellulose requirement huge, supply falls short of demand already to make limited Sucus Cocois resource, and be that main raw material has region restriction with Sucus Cocois, the needs of market amplification cannot be met.Find the large focus that the bacteria cellulose raw material substituting Sucus Cocois becomes research.The present inventor place seminar early-stage Study shows, inulin can be used for culturing bacterium Mierocrystalline cellulose, but during with pure inulin (inulin substratum base-material) fermented bacterial cellulose, output is only 0.26g/L, and output is far below the 4.80g/L of basic medium (chemosynthesis substratum).The present invention is directed to the problem that inulin culturing bacterium Mierocrystalline cellulose dry film productive rate is low, with conventional acetobacter xylinum for test strain, inulin substratum base-material is optimized, the bacteria cellulose that cultivated bacteria cellulose and basic medium ferment is compared in output and product performance, to finding a kind of method improving bacteria cellulose dry film productive rate, quality.
Summary of the invention
The object of the invention is to produce bacteria cellulose for current Sucus Cocois and have that the production cycle grow, yields poorly, labour intensity is large, production cost is high, seasonal and region limits by raw material problem, study and replace Sucus Cocois with other raw materials, produce that output is high, the measured bacteria cellulose of matter.Specifically utilizing inulin to replace Sucus Cocois, improving the method for bacteria cellulose dry film productive rate and quality by optimizing inulin fermention medium.
The present invention's one is optimized inulin fermention medium and is improved bacteria cellulose dry film productive rate, the method of quality, by adopting Plackett-Burman test design method, main ingredient in the inulin fermention medium of fermented bacterial cellulose is screened, effect significant factor is optimized further as key factor, inquire into the optimal proportion of each component of inulin fermention medium, obtaining the inulin fermentative medium formula proportioning optimized is: yeast extract paste 10g, peptone 8.0g, potassium primary phosphate 4.3g, magnesium sulfate 2.5g, 1000ml is made into inulin substratum base-material, 121 DEG C of sterilizing 20-30min, dehydrated alcohol 30ml is sterilely added after sterilizing cooling, be 5.54g/L with the productive rate of the inulin fermention medium fermented bacterial cellulose optimized, it is 21.31 times of inulin substratum base-material fermented bacterial cellulose dry film productive rate 0.26g/L, 15.42% is improve than basic medium fermented bacterial cellulose dry film productive rate 4.80g/L, primary quality measure is better than the bacteria cellulose of base culture base.
Described Plackett-Burman test design method is with peptone, yeast extract paste, KH
2pO
4, MgSO
4, FeSO
4, 7 factors testing as Plackett-Burman of citric acid, dehydrated alcohol 7 kinds of compositions, each factor gets high and low 2 levels, with bacteria cellulose dry film productive rate g/L for response value, carry out 12 groups of tests according to Plackett-Burman test design, the factor obtaining wherein remarkably influenced is dehydrated alcohol.
Effect significant factor is optimized further as key factor, that the dehydrated alcohol of remarkably influenced bacteria cellulose dry film productive rate is optimized, by investigating 2.2%-4.0%v/v dehydrated alcohol addition to the impact of bacteria cellulose dry film productive rate, find that bacteria cellulose dry film productive rate presents with the increase of dehydrated alcohol addition the process first increasing and reduce afterwards.When dehydrated alcohol addition is 3.0%v/v, bacteria cellulose dry film productive rate reaches maximum.
The preparation method of above-mentioned indication inulin substratum base-material is: the total sugar content surveying inulin through DNS method is 78.82g/100g inulin, and be dissolved in 1000ml distilled water by this inulin 63.5g, gained inulin solution total sugar content is 50g/L.
Above-mentioned indication basal medium formulation is: sucrose 50g, yeast extract paste 10g, peptone 8.0g, potassium primary phosphate 4.3g, magnesium sulfate 2.5g, is made into 1000ml with distilled water, add dehydrated alcohol 30ml after sterilizing cooling, utilize the output of basic medium fermented bacterial cellulose for 4.80g/L.
The bacteria cellulose primary quality measure of above-mentioned indication is retentiveness, degree of crystallinity, thermostability.
By the method for the inulin medium preparing bacteria cellulose optimized be:
(1) inulin fermention medium (abbreviation fermention medium) 1000ml of preparation optimization;
(2) 80ml is added through the cultivation acetobacter xylinum seed liquor of 24 hours;
(3) in 30 DEG C of constant incubator quiescent culture 8 days;
(4) 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;
(5) by the bacteria cellulose film 0.1mol/LHCl of step (4) and 30min, distilled water fully washs;
(6) step (5) is dried to constant weight at 80 DEG C to get product.
beneficial effect
The invention provides raw material based on inulin and pass through optimization, to optimize the method for inulin substratum fermented bacterial cellulose, the object improving bacteria cellulose dry film productive rate and quality can be reached, solve a difficult problem for region restriction in bacteria cellulose production, add the output of bacteria cellulose, reduce production cost, improve economic benefit.
Accompanying drawing explanation
Fig. 1 dehydrated alcohol addition is on the impact of bacteria cellulose dry film productive rate
The TG-DTA collection of illustrative plates of Fig. 2 basic medium bacteria cellulose
The TG-DTA collection of illustrative plates of Fig. 3 inulin substratum bacteria cellulose.
Embodiment
embodiment 1the preparation of inulin substratum base-material:
The total sugar content surveying inulin through DNS method is 78.82g/100g inulin, and be dissolved in 1000ml distilled water by this inulin 63.5g, gained inulin solution total sugar content is 50g/L.
embodiment 2the preparation of basic medium:
Get sucrose 50g, yeast extract paste 10g, peptone 8.0g, potassium primary phosphate 4.3g, magnesium sulfate 2.5g, is made into 1000mL with distilled water, adds dehydrated alcohol 3%v/v after sterilizing cooling.
embodiment 3with inulin substratum base-material fermented bacterial cellulose:
(1) inulin fermention medium base-material 1000ml is prepared;
(2) 80ml acetobacter xylinum seed liquor is added;
(3) in 30 DEG C of constant incubator quiescent culture 8 days;
(4) 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;
(5) by the bacteria cellulose film 0.1mol/LHCl of step (4) and 30min, distilled water fully washs;
(6) step (5) is dried to constant weight at 80 DEG C to get product.
Utilize inulin substratum base-material fermented bacterial cellulose dry film productive rate to be 0.26g/L, be far smaller than the output of basic medium 4.80g/L.Due to the total sugar content of inulin substratum base-material and the identical of basic medium, be all 50g/L, illustrate and cause the not high reason of inulin substratum base-material fermented bacterial cellulose dry film productive rate to be other nutritive ingredients in addition to sugars.
embodiment 4plackett-Burman test design
On the basis of single factor experiment, with peptone, yeast extract paste, KH
2pO
4, MgSO
4, FeSO
4, 7 factors testing as Plackett-Burman of citric acid, dehydrated alcohol 7 kinds of compositions, each factor gets high and low 2 levels, and level of factor is in table 1.With bacteria cellulose dry film productive rate R(g/L dry film weight) for response value, carry out 12 groups of tests according to Plackett-Burman test design, Plackett-Burman test design and response value are in table 2.DesignExpert7.0 is utilized to analyze Plackett-Burman testing data, as table 3 after arranging.
Through software analysis main effect, dehydrated alcohol (
p=0.0002) effect remarkable (see table 3),
pvalue is less than 0.10, can as the key factor optimized further.Other factors to Influence on test result little (
p>0.10), consider as condition element in studying further.
Note: * represents the difference in 0.10 level
embodiment 5the determination of key factor dehydrated alcohol addition:
Tested from Plackett-Burman, in 7 detection factors, dehydrated alcohol is the most significant influence factor, other factors are little to Influence on test result, determine that the addition of other factors is high level and the mean value of low-level addition, namely yeast extract paste is 12.5g/L, peptone is 10.0g/L, KH
2pO
4for 6.5g/L, MgSO
4for 3.1g/L, FeSO
4for 0.3g/L, citric acid are 0.3g/L.Investigate 2.2%-4.0%v/v dehydrated alcohol addition to the impact of bacteria cellulose dry film productive rate, the results are shown in Figure 1.As shown in Figure 1, bacteria cellulose dry film productive rate presents with the increase of dehydrated alcohol addition the process first increasing and reduce afterwards.When dehydrated alcohol addition is 3.0%v/v, bacteria cellulose dry film productive rate reaches maximum 5.54g/L, is 21.31 times of (0.26g/L) before not optimizing.
embodiment 6 is usedoptimize inulin fermention medium and produce bacteria cellulose:
(1) inulin fermention medium (abbreviation fermention medium) 1000ml of preparation optimization;
(2) 80ml is added through the cultivation acetobacter xylinum seed liquor of 24 hours;
(3) in 30 DEG C of constant incubator quiescent culture 8 days;
(4) 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;
(5) by the bacteria cellulose film 0.1mol/LHCl of step (4) and 30min, distilled water fully washs;
(6) step (5) is dried to constant weight at 80 DEG C to get product.
embodiment 7optimize inulin fermention medium and produce bacteria cellulose Performance Detection
(1) bacteria cellulose retentiveness and rehydration analysis
Bacteria cellulose retentiveness and rehydration analytical results are in table 4.As shown in Table 4, compared with basic medium, the bacteria cellulose of inulin culture medium culturing in retentiveness and rehydration a little more than the bacteria cellulose of base culture base.
(2) bacteria cellulose crystallinity analysis
According to X-ray diffraction experimental data, respectively according to the degree of crystallinity of each substratum bacteria cellulose of formulae discovery, the results are shown in Table 5.Compared with basic medium, the bacteria cellulose degree of crystallinity 83.99% of inulin substratum fermentation, improves 10.12% than 76.27% of basic medium.
(3) bacteria cellulose thermal stability analysis
The TG-DTA collection of illustrative plates of the bacteria cellulose of substratum and the fermentation of inulin substratum based on Fig. 2, Fig. 3, from the bacteria cellulose TG curve of Fig. 2, Fig. 3, the quality of institute's test sample product varies with temperature and can be divided into three phases: loss that the quality in 0 ~ 170 DEG C is slight, the quality rapid deterioration stage in 170 ~ 550 DEG C, the slight loss stage of the quality between 550 ~ 800 DEG C.65.1 DEG C and 46.9 DEG C are respectively in the maximum weight loss rate temperature of the bacteria cellulose of 0 ~ 170 DEG C of stage inulin substratum, basic medium gained, the mass loss showing this stage mainly bacteria cellulose dehydration produces, therefore claim the dehydration stage, due to institute's test sample product do not reach constant weight drying degree or because absorb moisture in air, so the temperature causing maximum weight loss rate place is different; 170 ~ 550 DEG C of stages, maximum weight loss rate place temperature is respectively 347.5 DEG C, 333.5 DEG C, this stage quality underspeeds soon, time length is long, for the degradation period of bacteria cellulose, adopt maximum weight loss rate method to weigh for material thermostability, maximum weight loss rate place temperature is higher, the thermostability of bacteria cellulose is better, illustrates that thermostability aspect inulin substratum is better than basic medium.
Claims (7)
1. optimize inulin fermention medium raising bacteria cellulose dry film productive rate for one kind, the method of quality, it is characterized in that by adopting Plackett-Burman test design method, main ingredient in the inulin fermention medium of fermented bacterial cellulose is screened, effect significant factor is optimized further as key factor, inquire into the optimal proportion of each component of inulin fermention medium, obtaining the inulin fermentative medium formula proportioning optimized is: yeast extract paste 10g, peptone 8.0g, potassium primary phosphate 4.3g, magnesium sulfate 2.5g, 1000ml is made into inulin substratum base-material, 121 DEG C of sterilizing 20-30min, dehydrated alcohol 30ml is sterilely added after sterilizing cooling, be 5.54g/L with the productive rate of the inulin fermention medium fermented bacterial cellulose dry film optimized, it is 21.31 times of inulin substratum base-material fermented bacterial cellulose dry film productive rate 0.26g/L, 15.42% is improve than basic medium fermented bacterial cellulose dry film productive rate 4.80g/L, primary quality measure is better than the bacteria cellulose of base culture base.
2. a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality according to claims 1, is characterized in that described Plackett-Burman test design method is with peptone, yeast extract paste, KH
2pO
4, MgSO
4, FeSO
4, 7 factors testing as Plackett-Burman of citric acid, dehydrated alcohol 7 kinds of compositions, each factor gets high and low 2 levels, with bacteria cellulose dry film productive rate g/L for response value, carry out 12 groups of tests according to Plackett-Burman test design, the factor obtaining wherein remarkably influenced is dehydrated alcohol.
3. a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality according to claims 1, it is characterized in that described being optimized further as key factor by effect significant factor, that the dehydrated alcohol of remarkably influenced bacteria cellulose dry film productive rate is optimized, by investigating 2.2%-4.0%v/v dehydrated alcohol addition to the impact of bacteria cellulose dry film productive rate, find that bacteria cellulose dry film productive rate presents with the increase of dehydrated alcohol addition the process first increasing and reduce afterwards;
When dehydrated alcohol addition is 3.0%v/v, bacteria cellulose dry film productive rate reaches maximum.
4. a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality according to claims 1, it is characterized in that the preparation method of indication inulin substratum base-material is: the total sugar content surveying inulin through DNS method is 78.82g/100g inulin, be dissolved in 1000ml distilled water by this inulin 63.5g, gained inulin solution total sugar content is 50g/L.
5. a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality according to claims 1, it is characterized in that indication basal medium formulation is: sucrose 50g, yeast extract paste 10g, peptone 8.0g, potassium primary phosphate 4.3g, magnesium sulfate 2.5g, is made into 1000ml with distilled water, add dehydrated alcohol 30ml after sterilizing cooling, utilize the output of basic medium fermented bacterial cellulose for 4.80g/L.
6. a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality according to claims 1, is characterized in that the bacteria cellulose primary quality measure of indication is retentiveness, degree of crystallinity, thermostability.
7. a kind of method optimizing inulin fermention medium raising bacteria cellulose dry film productive rate, quality according to claims 1, is characterized in that the method for the inulin medium preparing bacteria cellulose with optimization is:
(1) inulin fermention medium (abbreviation fermention medium) 1000ml of preparation optimization;
(2) 80ml is added through the cultivation acetobacter xylinum seed liquor of 24 hours;
(3) in 30 DEG C of constant incubator quiescent culture 8 days;
(4) 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;
(5) by the bacteria cellulose film 0.1mol/LHCl of step (4) and 30min, distilled water fully washs;
(6) step (5) is dried to constant weight at 80 DEG C to get product.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242166A (en) * | 2011-06-28 | 2011-11-16 | 东华大学 | Preparation method of Bacterial Cellulose (BC) with carbon source of inulin |
| CN102250983A (en) * | 2011-06-28 | 2011-11-23 | 东华大学 | Method for preparing bacterial cellulose by taking Jerusalem artichoke as carbon source |
| CN104372048A (en) * | 2014-12-04 | 2015-02-25 | 贵州大学 | Method for increasing yield of bacterial cellulose produced by fermenting acetobacter xylinum |
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2015
- 2015-12-16 CN CN201510942263.9A patent/CN105368896A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242166A (en) * | 2011-06-28 | 2011-11-16 | 东华大学 | Preparation method of Bacterial Cellulose (BC) with carbon source of inulin |
| CN102250983A (en) * | 2011-06-28 | 2011-11-23 | 东华大学 | Method for preparing bacterial cellulose by taking Jerusalem artichoke as carbon source |
| CN104372048A (en) * | 2014-12-04 | 2015-02-25 | 贵州大学 | Method for increasing yield of bacterial cellulose produced by fermenting acetobacter xylinum |
Non-Patent Citations (1)
| Title |
|---|
| 李艳等: "利用菠萝酒合成细菌纤维素的发酵培养基优化", 《西北农业学报》 * |
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