CN102875641B - Method for continuously preparing pumpkin protein, ethanol fuel and dietary fiber - Google Patents
Method for continuously preparing pumpkin protein, ethanol fuel and dietary fiber Download PDFInfo
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Abstract
The invention relates to a method for continuously preparing pumpkin protein, ethanol fuel and dietary fiber, belonging to a technique for preparing active substances from natural substances. The method comprises the following steps: oscillating a pumpkin pulverized material solution at constant temperature, centrifuging, and filtering to obtain a filtrate and a filter residue; regulating the pH value of the filtrate to a protein isoelectric point by using a buffer solution, carrying out centrifugal separation to obtain a precipitate and a supernatant, carrying out washing and freeze-drying on the precipitate to obtain the pumpkin protein, and carrying out concentration and alcohol precipitation on the supernatant to obtain the water-soluble dietary fiber; and washing the pumpkin filter residue with water, adding water, sequentially carrying out alpha-amylase liquification and saccharifying enzyme saccharification, carrying out centrifugal separation to obtain a precipitate and a supernatant, washing the precipitate with water, carrying out freeze-drying to obtain the water-insoluble dietary fiber, carrying out fermentation treatment on the supernatant with active dry yeast, filtering, distilling the filtrate, and dehydrating with a desiccant to obtain the ethanol fuel. The invention has the advantages of no mutual interference of various steps, high product yield, simple technological operation and low cost, and can easily implement industrial production.
Description
Technical field
The invention belongs to the technology for preparing active substance from crude substance, relate generally to a kind of method that the discarded pulp of seed-use pumpkin is raw material continuous production Cucurmosin, alcohol fuel and food fibre of take.
Background technology
Pumpkin (
Cucurbita moschataDuch) be the Curcurbitaceae vine pumpkin fruit of overgrowing.China's pumpkin aboundresources, the pumpkin planting area occupies second place of the world, and ultimate production ranks first in the world, and wherein 20% is edible, and 80% is seed-use pumpkin.Pumpkin nutrient is worth very high, be rich in the multiple nutritional components such as amino acid, protein, Mierocrystalline cellulose, electrolytes and minerals, also contain polysaccharide, carotenoid etc. and there is the nourishing function bioactive ingredients, be mainly used in antitumor, hypoglycemic, reducing blood-fat, prevention prostatosis and strengthen the medical field such as immunity.Along with to the going deep into of pumpkin nutrient composition and value medical health care research, the exploitation of pumpkin product is subject to the attention of countries in the world day by day, and very wide exploitation prospect is arranged.
Seed-use pumpkin is the important pumpkin Cultivar of China, and because the nutrient composition contents such as solubility sugar and starch in the pulp of seed-use pumpkin kind are on the low side, mouthfeel is bad, and pulp is basic all discarded, causes serious waste.How the discarded pulp of seed-use pumpkin kind is taken full advantage of to become in the seed-use pumpkin variety production and be badly in need of the subject matter solved.In seed-use pumpkin pulp, protein content, in 1.5% left and right, due to less to the research of pumpkin fruit albumen, therefore can be used as new protein resource research and development both at home and abroad; Food fibre and food thereof as food function factor, the world in recent years various countries have also obtained paying close attention to widely and development rapidly, in seed-use pumpkin pulp, dietary fiber content, up to 12.4%, is the main component of dry-matter, develops the seed-use pumpkin food fibre and has huge market; In seed-use pumpkin pulp, starch content is lower, and content is in 3% left and right, but discarded pulp cost is zero, can be used as the raw material for preparing alcohol fuel, alleviates because grain prepares grain that ethanol brings and the rise of oil price.Therefore, the discarded pulp of the seed-use pumpkin of take is raw material, and continuous production Cucurmosin, alcohol fuel and food fibre have wide market, and its prospect is very considerable.
At present about the preparation of activeconstituents in pumpkin fruit with utilize aspect to have some Patents, as domestic patent CN102180991A " a kind of method that fully utilizes pumpkin ", disclose a kind of substep extraction method and extracted and utilized the membrane separation technique concentrated method that removed the gred, produce pectin, squash polyoses and pumpkin powder, but do not relate to the continuous production of other active substances such as albumen, food fibre in pumpkin fruit; Patent CN102524698A " a kind of preparation method of pumpkin dietary fiber ", disclose and a kind ofly utilized ultrafiltration, steaming and decocting under high pressure to obtain the method for pumpkin dietary fiber in conjunction with high-pressure homogeneous and vacuum microwave treatment process; Patent CN101195835A " utilizes the preparation technology of pumpkin for fermenting raw materials ethanol ", relates to and take pumpkin fruit as raw material, smashes the method that obtains ethanol after defibrination by enzymatic hydrolysis and fermentation and second distillation.But patent CN102524698A only can separate and utilize a kind of effective constituent with CN101195835A, and other activeconstituentss have to remove as impurity in purge process, has caused the very large wasting of resources.
Summary of the invention
A kind of method that the purpose of this invention is to provide continuous production Cucurmosin, alcohol fuel and food fibre, the purpose that reach simple to operate, fully utilizes each active substance in pumpkin.
The object of the present invention is achieved like this:
A kind of method of continuous production Cucurmosin, alcohol fuel and food fibre, comprise the discarded pulp of seed-use pumpkin cleaned, remove the peel, dry, pulverized; 1. by discarded 1～5% NaOH solution pulp comminuting matter for of pumpkin, constant temperature oscillation, centrifugation, filtration under liquid ratio 20:1～30:1,60～80 ℃ of temperature of extraction, extraction times 50～70 min conditions, obtain filtrate and filter residue; 2. in gained filtrate, add damping fluid to regulate the pH value to isoelectric points of proteins, centrifugation, must precipitate and supernatant liquor, and washing of precipitate, lyophilize are obtained to Cucurmosin; 3. step 2. in centrifugation gained supernatant liquor obtain water-soluble dietary fibre through concentrated, alcohol precipitation; 4. after step, 1. middle filtration gained filter residue washes with water, by liquid ratio, 3:1～8:1 adds water, successively with α-amylase insulation liquefaction and saccharifying enzyme insulation saccharification, centrifugation, must precipitate and supernatant liquor, by precipitation wash with water, lyophilize obtains water insoluble dietary fiber; 5. 4. active dry yeast constant temperature oscillation fermentation for centrifugation gained supernatant liquor of step, the gained fermented liquid obtains alcohol fuel after distillation, desiccant dehydration.
Beneficial effect of the present invention is, selects 1～5% NaOH solution to have good lixiviate ability to Cucurmosin, and to insoluble dietary fibre in pumpkin powder and starch lixiviate ability; In addition, by Cucurmosin and alcohol fuel preparation condition are carried out to response surface optimization, protein extracting ratio and ethanol fermentation rate all reach the highest, under top condition, Cucurmosin product protein content reaches 70～80%, under optimum enzymolysis condition, the ethanol fermentation rate reaches 65～75%, then obtains the alcohol fuel that content is 90～99% after single flash, desiccant dehydration.It is raw material that the discarded pulp of seed-use pumpkin is take in the present invention, and continuous, easy production has high value-added product, can pull economy, drives farmer richness.This method is simple to operate, does not need complicated equipment, is easy to industrialization, has broad application prospects.
Embodiment
Below the embodiment of the present invention is described in detail.
A kind of method of continuous production Cucurmosin, alcohol fuel and food fibre, comprise the discarded pulp of seed-use pumpkin cleaned, remove the peel, dry, pulverized; 1. by discarded 1～5% NaOH solution pulp comminuting matter for of pumpkin, constant temperature oscillation, centrifugation, filtration under liquid ratio 20:1～30:1,60～80 ℃ of temperature of extraction, extraction times 50～70 min conditions, obtain filtrate and filter residue; 2. in gained filtrate, add damping fluid to regulate the pH value to isoelectric points of proteins, centrifugation, must precipitate and supernatant liquor, and washing of precipitate, lyophilize are obtained to Cucurmosin; 3. step 2. in centrifugation gained supernatant liquor obtain water-soluble dietary fibre through concentrated, alcohol precipitation; 4. after step, 1. middle filtration gained filter residue washes with water, by liquid ratio, 3:1～8:1 adds water, successively with α-amylase insulation liquefaction and saccharifying enzyme insulation saccharification, centrifugation, must precipitate and supernatant liquor, by precipitation wash with water, lyophilize obtains water insoluble dietary fiber; 5. 4. active dry yeast constant temperature oscillation fermentation for centrifugation gained supernatant liquor of step, the gained fermented liquid obtains alcohol fuel after distillation, desiccant dehydration.
The step damping fluid that 2. the middle pH of adjusting value is used to isoelectric points of proteins is 0.1mol/L acetic acid-sodium acetate solution, and the Cucurmosin isoelectric pH value is in 3 ~ 6 scopes.
Step 2. middle centrifugation rotating speed is 5000～10000r/min, in centrifugation time 10～40min scope.
Step 2. middle washing precipitation adopts acetone and 95% ethanol continuous washing.
The step condition that 3. middle supernatant liquor prepares water-soluble dietary fibre is pH value 6～8,2～6 times of supernatant liquor volumes of dehydrated alcohol consumption, alcohol precipitation times 30～70 min.
Step 4. middle α-amylase insulation liquefaction top condition is pH value 4～6, α-amylase addition 0.05～0.15%, 60～80 ℃ of liquefaction temperatures, liquefying time 50～70 min.
Step 4. middle saccharifying enzyme insulation saccharification top condition is pH value 3～4, saccharifying enzyme addition 0.05～0.15%, 55～65 ℃ of saccharification temperatures, saccharification time 10～20 min.
Step 5. middle supernatant liquor is pH value 4～5, inoculum of dry yeast 0.05～0.10%, 28～30 ℃ of leavening temperatures, fermentation time 65～75 h by active dry yeast constant temperature oscillation fermentation top condition.
When step is prepared into alcohol fuel by fermented liquid in 5., the dehydration siccative that uses is unslaked lime.
Determining of Cucurmosin optimum extraction condition
Determining of seed-use pumpkin pulp albumen iso-electric point:
Albumen after extracting is dissolved in to the HAc-NaAc buffer system, regulate pH value 2 ~ 8, centrifugal 10～the 40min of 5000～10000r/min, obtain protein precipitation, with each washing precipitation of ethanol of acetone and 95% 3 times, take quality after lyophilize, according to precipitation quality judging pumpkin fruit albumen iso-electric point, determine that seed-use pumpkin pulp albumen iso-electric point is 3 ~ 6 in the pH value.
The impact of liquid ratio on extraction rate of protein:
Get the 10.00g pumpkin powder, add 3%NaOH solution by different liquid ratios respectively, 80 ℃ of water-bath 1h, centrifugal suction filtration, supernatant liquor adjusts pH to isoelectric points of proteins, protein precipitation is separated out, centrifugal, the washing with alcohol of acetone and 95% for precipitation, last lyophilize, obtain pumpkin fruit albumen, and calculate the extraction yield of protein.Determine that liquid ratio is in 10:1～30:1 scope.
The impact of NaOH massfraction on extraction rate of protein:
Get the 10.00g pumpkin powder, in liquid ratio, be that the 20:1 ratio adds respectively different concns NaOH solution, 80 ℃ of water-bath 1h, centrifugal suction filtration, supernatant liquor adjusts pH to isoelectric points of proteins, protein precipitation is separated out, centrifugal, the washing with alcohol of acetone and 95% for precipitation, last lyophilize, obtain pumpkin fruit albumen, and calculate the extraction yield of protein.Determine that the NaOH liquid quality fraction is 1～5%.
The impact of alkali solubility temperature on extraction rate of protein:
Get the 10.00g pumpkin powder, in liquid ratio, be that the 20:1 ratio adds 3%NaOH solution, water-bath 1h under differing temps respectively, centrifugal suction filtration, supernatant liquor adjusts pH to isoelectric points of proteins, protein precipitation is separated out, centrifugal, the washing with alcohol of acetone and 95% for precipitation, last lyophilize, obtain pumpkin fruit albumen, and calculate the extraction yield of protein.Determine that extracting temperature is 50～90 ℃.
The impact of molten time of alkali on extraction rate of protein:
Get the 10.00g pumpkin powder, in liquid ratio, be that the 20:1 ratio adds 3%NaOH solution, difference water-bath different time under 80 ℃, centrifugal suction filtration, supernatant liquor adjusts pH to isoelectric points of proteins, protein precipitation is separated out, centrifugal, the washing with alcohol of acetone and 95% for precipitation, last lyophilize, obtain pumpkin fruit albumen, and calculate the extraction yield of protein.Determine that extraction time is 15～75 min.
Second-order regression for rotatable orthogonal design (1/2 implements):
Take extraction rate of protein mean value as examination index (establishing 3 repetitions), according to the single factor experiment result, select liquid ratio, NaOH concentration, temperature, time, these four factors were carried out the quadratic regression orthogonal rotational test, utilized SAS software to set up regression equation to test-results, and experimental factor is carried out to the pairwise interaction analysis.
Table 1 quadratic regression rotary combination design table and extraction yield (1/2 implements)
The every variance analysis of table 2 regression equation
| The regression variance source | Degree of freedom | Sum of squares | Inequality | The F value | Prob﹥F | Significance |
| Once | 4 | 227.503591 | 0.5877 | 56.20 | <.0001 | ** |
| Quadratic term | 4 | 92.336065 | 0.2385 | 22.81 | 0.0009 | ** |
| Mutual | 6 | 61.182868 | 0.1581 | 10.08 | 0.0064 | * |
| Regression model | 14 | 381.022524 | 0.9843 | 26.89 | 0.0003 | ** |
| Lose and intend item | 2 | 4.428377 | 2.214189 | 5.39 | 0.0733 | ? |
| Pure error | 4 | 1.643680 | 0.410920 | ? | ? | ? |
| Total error | 6 | 6.072057 | 1.012010 | ? | ? | ? |
| Population variance | 20 | 387.094581 | ? | ? | ? | ? |
Table 3 regression analysis coefficient value and analytical results
| Parameter | Regression coefficient | Standard deviation | The T value | Prob﹥T | Significance |
| Constant | -257.199519 | 73.484971 | -3.50 | 0.0128 | * |
| b | 3.900360 | 2.367987 | 1.65 | 0.1506 | ? |
| c | -47.330732 | 11.909348 | -3.97 | 0.0073 | * |
| t | 5.746751 | 1.306707 | 4.40 | 0.0046 | ** |
| h | 4.547766 | 0.793957 | 5.73 | 0.0012 | ** |
| b*b | -0.118520 | 0.029300 | -4.05 | 0.0068 | ** |
| c*b | 1.457917 | 0.308652 | 4.72 | 0.0032 | ** |
| c*c | -2.363003 | 0.732488 | -3.23 | 0.0180 | * |
| t*b | -0.004236 | 0.019759 | -0.21 | 0.8373 | ? |
| t*c | 0.265625 | 0.098797 | 2.69 | 0.0361 | * |
| t*t | -0.032430 | 0.007325 | -4.43 | 0.0044 | ** |
| h*b | -0.059454 | 0.020577 | -2.89 | 0.0277 | ** |
| h*c | 0.338565 | 0.102884 | 3.29 | 0.0166 | * |
| h*t | -0.022523 | 0.006586 | -3.42 | 0.0142 | * |
| h*h | -0.024747 | 0.003256 | -7.60 | 0.0003 | ** |
Extraction yield under table 4 optimal conditions under optimal value and optimal conditions
Carry out proof test, triplicate according to physical condition at optimum extraction conditions (liquid ratio 20:1～30:1, NaOH concentration 1～5%, 60～80 ℃ of temperature, time 50～70min).It is 70～80% that result obtains extraction rate of protein, and trial value and model theory value are very approaching.And the revision test relative deviation is no more than 1～2%, illustrate that proteins extraction condition circulation ratio is good.
The alcohol fuel preparation condition is optimized
Liquefaction condition is optimized:
In the molten condition of the suitableeest alkali, taking the 20.00g pumpkin powder, to carry out alkali molten, and centrifugation, wash the molten residue obtained of alkali with water, then adds suitable quantity of water, after adjusting the temperature to liquefaction temperature, adds α-amylase, and insulation is liquefied.Solution after liquefaction is cooled to 60 ℃, by 0.1% of raw materials quality, adds the activation saccharifying enzyme, water bath heat preservation 10min carries out saccharification.Centrifugation after saccharification finishes, be cooled to 30 ℃ by supernatant liquor, by 0.1% of raw materials quality, adds the activation yeast, the jam-pack fermentation bung, and 48h ferments in constant-temperature shaking incubator.Measure CO
2Weight loss and alcoholic strength, with how many definite each factor optimum values of fermentation rate.
PH affects the ethanol fermentation rate:
Determine that the α-amylase consumption is 0.1% of raw material, 70 ℃ of liquefaction temperatures, liquefying time is 60min, regulates respectively different pH.Take the ethanol fermentation rate as index, determine that the pH value is 4～8.
The impact of α-amylase addition on the ethanol fermentation rate:
Determine that liquefying time is 60min, pH is 6, and liquefaction temperature is 70 ℃, adds the different mass α-amylase by raw material respectively.Take the ethanol fermentation rate as index, determine that the α-amylase addition is 0.01～0.2%.
Liquefaction temperature affects the ethanol fermentation rate:
Determine that liquefying time is 60min, pH is 6, and the α-amylase consumption is raw material 0.1%, regulates respectively different liquefaction temperatures 5.Take the ethanol fermentation rate as index, determine that liquefaction temperature is 50～90 ℃.
Liquefying time affects the ethanol fermentation rate:
Determine that pH is 6, liquefaction temperature is 70 ℃, and the α-amylase consumption is raw material 0.1%, regulates respectively different liquefying times.Take the ethanol fermentation rate as index, determine that liquefying time is 30～70 min.
Second-order regression for rotatable orthogonal design:
According to the single factor experiment result, take fermentation rate mean value as examination index (establishing 3 repetitions), select pH, α-amylase addition, liquefaction temperature, liquefying time to carry out four factors quadratic regression orthogonal rotational tests, experimental factor has been carried out to the pairwise interaction analysis.
According to the single factor experiment result, take fermentation rate mean value as examination index (establishing 3 repetitions), select pH, α-amylase addition, liquefaction temperature, liquefying time to carry out four factors quadratic regression orthogonal rotational tests, experimental factor has been carried out to the pairwise interaction analysis.
Table 5 quadratic regression rotary combination design table and fermentation rate (1/2 implements)
The every variance analysis of table 6 regression equation
| The regression variance source | Degree of freedom | Sum of squares | Inequality | The F value | Prob﹥F | Significance |
| Once | 4 | 176.004811 | 0.6547 | 135.43 | <.0001 | ** |
| Quadratic term | 4 | 76.992190 | 0.2864 | 59.24 | <.0001 | ** |
| Mutual | 6 | 13.897576 | 0.0517 | 7.13 | 0.0153 | * |
| Regression model | 14 | 266.894577 | 0.9927 | 58.68 | <.0001 | ** |
| Lose and intend item | 2 | 1.111423 | 0.555712 | 2.65 | 0.1848 | ? |
| Pure error | 4 | 0.838000 | 0.209500 | ? | ? | ? |
| Total error | 6 | 1.949423 | 0.324904 | ? | ? | ? |
| Population variance | 20 | 268.844 | ? | ? | ? | ? |
Table 7 regression analysis coefficient value and analytical results
| Parameter | Regression coefficient | Standard deviation | The T value | Prob﹥T | Significance |
| Constant | -368.134753 | 51.094621 | -7.20 | 0.0004 | ** |
| p | 43.243843 | 7.905475 | 5.47 | 0.0016 | ** |
| c | 354.277472 | 161.140931 | 2.20 | 0.0702 | ? |
| t | 3.379120 | 0.736191 | 4.59 | 0.0037 | ** |
| h | 5.688788 | 0.795790 | 7.15 | 0.0004 | ** |
| p*p | -4.025413 | 0.415036 | -9.70 | <.0001 | ** |
| c*p | 44.319444 | 17.488592 | 2.53 | 0.0444 | * |
| c*c | -1402.165138 | 166.014296 | -8.45 | 0.0002 | ** |
| t*p | 0.026736 | 0.055980 | 0.48 | 0.6498 | ? |
| t*c | 2.909722 | 1.119592 | 2.60 | 0.0407 | * |
| t*t | -0.025154 | 0.004150 | -6.06 | 0.0009 | ** |
| h*p | -0.080347 | 0.087443 | -0.92 | 0.3936 | ? |
| h*c | -9.270833 | 1.748859 | -5.30 | 0.0018 | ** |
| h*t | -0.003646 | 0.005598 | -0.65 | 0.5390 | ? |
| h*h | -0.035604 | 0.004150 | -8.58 | 0.0001 | ** |
Fermentation rate under table 8 optimal conditions under optimal value and optimal conditions
Carry out proof test, triplicate according to physical condition at optimum extraction conditions (pH value 4～6, α-amylase addition 0.05～0.15%, 60～80 ℃ of temperature, time 50～70min).It is 50～60% that result obtains fermentation rate, and trial value and model theory value are very approaching.And the revision test relative deviation is no more than 0～1 %, illustrate that the liquefaction condition circulation ratio is good.
The saccharification condition optimizing:
According to the molten condition of the suitableeest alkali and the suitableeest liquefaction condition, take the 20.00g pumpkin powder and carry out that alkali is molten, liquefaction, after the saccharification temperature that the karusen that obtains after liquefaction is cooled to set, add the saccharifying enzyme after activation, water-bath vibration certain hour carries out saccharification.Centrifugation after saccharification finishes, be cooled to 30 ℃ by supernatant liquor, by 0.1% of raw materials quality, adds the activation yeast, the jam-pack fermentation bung, and 48h ferments in constant-temperature shaking incubator.Measure CO
2Weight loss and alcoholic strength, with how many definite each factor optimum values of fermentation rate.
The impact of saccharification pH on alcohol getting rate:
Determine that saccharification temperature is 60 ℃, saccharification 20min, add saccharifying enzyme by 0.1% of raw material, regulates respectively different pH.Take the ethanol fermentation rate as index, determine that the pH value is 3～5.
The impact of saccharifying enzyme addition on the ethanol fermentation rate:
Determine that pH is 4, saccharification temperature is 60 ℃, and saccharification 20min adds the different mass saccharifying enzyme by raw material respectively.Take the ethanol fermentation rate as index, determine that the saccharifying enzyme addition is 0.05～0.25%.
The impact of saccharification temperature on the ethanol fermentation rate:
Determine that pH is 4, by 0.1% of raw material, add saccharifying enzyme, saccharification 20min, regulate respectively different saccharification temperatures.Take the ethanol fermentation rate as index, determine that saccharification temperature is 50～70 ℃.
The impact of saccharification time on alcohol getting rate:
Determine that pH is 4, by 0.1% of raw material, add saccharifying enzyme, saccharification temperature is 60 ℃, respectively the saccharification different time.Take the ethanol fermentation rate as index, determine that saccharification time is 5～30 min.
Quadratic regression rotation composite test
According to the single factor experiment result, take fermentation rate mean value as examination index (establishing 3 repetitions), select pH, saccharifying enzyme addition, saccharification temperature, saccharification time to carry out four factors quadratic regression orthogonal rotational tests, experimental factor has been carried out to the pairwise interaction analysis.
Table 9 quadratic regression rotary combination design table and fermentation rate
The every variance analysis of table 10 regression equation
| The regression variance source | Degree of freedom | Sum of squares | Inequality | The F value | Prob﹥F | Significance |
| Once | 4 | 270.964299 | 0.7567 | 247.27 | <.0001 | ** |
| Quadratic term | 4 | 77.266882 | 0.2158 | 70.51 | <.0001 | ** |
| Mutual | 6 | 8.225415 | 0.0230 | 5.00 | 0.0354 | * |
| Regression model | 14 | 356.456597 | 0.9954 | 92.94 | <.0001 | ** |
| Lose and intend item | 2 | 0.510927 | 0.255464 | 0.90 | 0.4749 | ? |
| Pure error | 4 | 1.132800 | 0.283200 | ? | ? | ? |
| Total error | 6 | 1.643727 | 0.273955 | ? | ? | ? |
| Population variance | 20 | 358.100324 | ? | ? | ? | ? |
Table 11 regression analysis coefficient value and analytical results
| Parameter | Regression coefficient | Standard deviation | The T value | Prob﹥T | Significance |
| Constant | -642.810507 | 82.634686 | -7.78 | 0.0002 | ** |
| p | 75.939927 | 17.947273 | 4.23 | 0.0055 | ** |
| c | 787.577595 | 183.685900 | 4.29 | 0.0052 | ** |
| t | 16.137279 | 2.027221 | 7.96 | 0.0002 | ** |
| h | 2.886443 | 1.836859 | 1.57 | 0.1671 | ? |
| p*p | -11.493046 | 1.524429 | -7.54 | 0.0003 | ** |
| c*p | 62.305556 | 32.117862 | 1.94 | 0.1005 | ? |
| c*c | -1747.304644 | 152.442926 | -11.46 | <.0001 | ** |
| t*p | 0.118056 | 0.205614 | 0.57 | 0.5867 | ? |
| t*c | -8.597222 | 2.056135 | -4.18 | 0.0058 | ** |
| t*t | -0.132330 | 0.015244 | -8.68 | 0.0001 | ** |
| h*p | 0.033056 | 0.321179 | 0.10 | 0.9214 | ? |
| h*c | -8.380556 | 3.211786 | -2.61 | 0.0402 | ** |
| h*t | 0.026250 | 0.020561 | 1.28 | 0.2489 | ? |
| h*h | -0.122330 | 0.015244 | -8.02 | 0.0002 | ** |
Fermentation rate under table 12 optimal conditions under optimal value and optimal conditions
Carry out proof test, triplicate according to physical condition at optimum extraction conditions (pH value 3～4, saccharifying enzyme addition 0.05～0.15%, 55～65 ℃ of temperature, times 10～20 min).It is 60～70% that result obtains fermentation rate, and trial value and model theory value are very approaching.And the revision test relative deviation is no more than 0～1%, illustrate that saccharification condition circulation ratio is good.
Fermentation condition optimization:
According to the molten condition of the suitableeest alkali, the suitableeest liquefaction condition and the suitableeest saccharification condition, take that the 20.00g pumpkin powder carries out that alkali is molten, liquefaction, saccharification, by the karusen centrifugation after saccharification, after the leavening temperature that supernatant liquor is cooled to set, add the active dry yeast after activation, the jam-pack fermentation bung ferments in constant-temperature shaking incubator.After fermentation ends, measure CO
2Weight loss and alcoholic strength, with how many definite each factor optimum values of fermentation rate.
The impact of fermentation pH on the ethanol fermentation rate:
Determine that leavening temperature is 30 ℃, fermentation 48h, add yeast by 0.1% of raw material, regulates respectively different pH.Take the ethanol fermentation rate as index, determine that pH is 3.8～5.8.
The impact of inoculum of dry yeast on the ethanol fermentation rate:
Determine that fermentation pH is 4.8, leavening temperature is 30 ℃, and fermentation 48h, add the different mass yeast by raw material respectively.Take the ethanol fermentation rate as index, determine that inoculum of dry yeast is 0.01～0.2%.
The impact of leavening temperature on the ethanol fermentation rate:
Determine that fermentation pH is 4.8, fermentation 48h, add yeast by 0.1% of raw material, regulates respectively the different fermentations temperature.Take the ethanol fermentation rate as index, determine that leavening temperature is 28～32 ℃.
The impact of fermentation time on the ethanol fermentation rate:
Determine that fermentation pH is 4.8, leavening temperature is 30 ℃, by 0.1% of raw material, adds yeast, and different time ferments respectively.Take the ethanol fermentation rate as index, determine that fermentation time is 40～80 h.
Second-order regression for rotatable orthogonal design
According to the single factor experiment result, take fermentation rate mean value as examination index (establishing 3 repetitions), select pH, inoculum of dry yeast, leavening temperature, fermentation time to carry out four factors quadratic regression orthogonal rotational tests, experimental factor has been carried out to the pairwise interaction analysis.
Table 13 quadratic regression rotary combination design table and fermentation rate
The every variance analysis of table 14 regression equation
| The regression variance source | Degree of freedom | Sum of squares | Inequality | The F value | Prob﹥F | Significance |
| Once | 4 | 312.202406 | 0.7322 | 243.30 | <.0001 | ** |
| Quadratic term | 4 | 95.753794 | 0.2246 | 74.62 | <.0001 | ** |
| Mutual | 6 | 16.508011 | 0.0387 | 8.58 | 0.0097 | * |
| Regression model | 14 | 424.464211 | 0.9955 | 94.51 | <.0001 | ** |
| Lose and intend item | 2 | 0.762449 | 0.381225 | 1.31 | 0.3647 | ? |
| Pure error | 4 | 1.162320 | 0.290580 | ? | ? | ? |
| Total error | 6 | 1.924769 | 0.320795 | ? | ? | ? |
| Population variance | 20 | 426.38898 | ? | ? | ? | ? |
Table 15 regression analysis coefficient value and analytical results
| Parameter | Regression coefficient | Standard deviation | The T value | Prob﹥T | Significance |
| Constant | -3031.604245 | 417.247599 | -7.27 | 0.0003 | ** |
| p | 96.347120 | 37.776469 | 2.55 | 0.0435 | * |
| c | 773.463429 | 380.640319 | 2.03 | 0.0884 | ? |
| t | 175.109986 | 25.448957 | 6.88 | 0.0005 | ** |
| h | 7.423527 | 1.607719 | 4.62 | 0.0036 | ** |
| p*p | -19.253003 | 1.650239 | -11.67 | <.0001 | ** |
| c*p | 92.768519 | 34.471246 | 2.69 | 0.0360 | * |
| c*c | -1677.300322 | 165.023897 | -10.16 | <.0001 | ** |
| t*p | 2.812500 | 1.112490 | 2.53 | 0.0448 | * |
| t*c | -13.541667 | 11.124903 | -1.22 | 0.2692 | ? |
| t*t | -3.028251 | 0.412560 | -7.34 | 0.0003 | ** |
| h*p | -0.243545 | 0.146853 | -1.66 | 0.1483 | ? |
| h*c | -7.980047 | 1.468534 | -5.43 | 0.0016 | ** |
| h*t | -0.094777 | 0.047007 | -2.02 | 0.0904 | ? |
| h*h | -0.021457 | 0.002882 | -7.45 | 0.0003 | ** |
Fermentation rate under table 16 optimal conditions under optimal value and optimal conditions
Carry out proof test, triplicate according to physical condition at optimum extraction conditions (pH value 4～5, inoculum of dry yeast 0.05～0.10%, 28～30 ℃ of temperature, times 65～75 h).It is 65～75% that result obtains fermentation rate, and trial value and model theory value are very approaching, and the revision test relative deviation is no more than 0～1 %, illustrates that the fermentation condition circulation ratio is good.
Continuous production gained Cucurmosin and food fibre part nutritive ingredient and physico-chemical property
Cucurmosin main nutrient composition and part physical and chemical index:
The Cucurmosin obtained after extraction, freeze-drying, be faint yellow, and quality is better, and wherein protein content reaches 70～80%, with the raw material pumpkin powder, compares, and food fibre and starch content obviously reduce.
Table 17 prepares the gained Cucurmosin and the raw material main nutrient composition compares (%)
| Sample | Protein | Food fibre | Starch |
| Pumpkin powder | 8.85 | 41.98 | 38.64 |
| Cucurmosin | 70.61±0.65 | 10.28±0.54 | 3.77±0.28 |
Table 18 prepares gained pumpkin protein isolate and the isolated soybean protein part physical and chemical index compares
| Sample | Retention ability (g/g) | Oil absorbency (g/g) | Viscosity (mPas) |
| Pumpkin | 2.38±0.04 | 2.07±0.05 | 4.2±0.1 |
| Soybean | 4 | 1.54 | 4.5 |
Known by contrasting, Cucurmosin has good physico-chemical property.At present, soybean protein isolate and soybean protein concentrate, in a large number for meat product, are exactly mainly the characteristics of utilizing its retentiveness good, reach the purpose of improving quality product, improving the product yield.The oil absorbency of protein plays very important effect in the food formulations such as meat product, milk-product and biscuit sandwich and processing.At beverage, soup, sauce and cream, in such fluid foodstuffs, protein system viscosity and denseness are critical function character, and viscosity is an important indicator of product, and the viscosity of protein is at the characteristic important role of adjusting these food.
Food fibre main nutrient composition and part physical and chemical index:
The water-soluble dietary fibre that extraction is obtained (SDF) and water insoluble dietary fiber (IDF), freeze-drying is pulverized to merge and is total dietary fiber (TDF).SDF is pale, and it is faint yellow that IDF is.Compare with the raw material pumpkin powder, prepare gained total dietary fiber content and reach 87.36%, compare with the raw material pumpkin powder, Protein and starch contents obviously reduces.
Table 19 prepares the gained food fibre and the raw material main nutrient composition compares (%)
| Sample | Protein | Food fibre | Starch |
| Pumpkin powder | 8.85 | 41.98 | 38.64 |
| Pumpkin dietary fiber | 0.34±0.02 | 87.36±0.73 | 2.47±0.16 |
Table 20 prepares the gained pumpkin dietary fiber and the Rhizoma Dioscoreae esculentae dietary fiber physico-chemical property compares
| Sample | Retention ability (g/g) | Bulging force (mL/g) | In conjunction with waterpower (g/g) | Oil absorbency (g/g) | Cation exchange capacity (CEC) (mmol/g) | Viscosity (mPas) |
| Pumpkin | 5.00±0.01 | 13±1 | 15.66±0.1 | 1.36±0.02 | 0.58±0.02 | 0.7±0.1 |
| Sweet potato | 8 | 8.5 | 10.4 | 2.4 | - | 1.3 |
By with Rhizoma Dioscoreae esculentae dietary fiber contrast, pumpkin dietary fiber has higher bulging force and in conjunction with waterpower, and retention ability is lower than Rhizoma Dioscoreae esculentae dietary fiber.Food fibre, by being combined with moisture, can improve the quality of bakery product; The high retentiveness of food fibre can increase volume and the speed of human body defecation, and toxic substance is excreted rapidly.Food fibre becomes large in conjunction with volume after water, causes satiety, and affects body and can utilize digesting and assimilating of carbohydrate etc. to other compositions of food, and obesity prevention benefits.Pumpkin dietary fiber absorbs the grease ability lower than sweet potato, but also can side direction reduces the secretion of human body bile acide and steroid.
Claims (7)
1. the method for a continuous production Cucurmosin, alcohol fuel and food fibre, comprise the discarded pulp of seed-use pumpkin cleaned, remove the peel, dry, pulverized; It is characterized in that: 1., by discarded 1～5% NaOH solution pulp comminuting matter for of pumpkin, constant temperature oscillation, centrifugation, filtration under liquid ratio 20:1～30:1,60～80 ℃ of temperature of extraction, extraction times 50～70 min conditions, obtain filtrate and filter residue; 2. in gained filtrate, add damping fluid to regulate the pH value to isoelectric points of proteins, centrifugation, must precipitate and supernatant liquor, and washing of precipitate, lyophilize are obtained to Cucurmosin; 3. step 2. in centrifugation gained supernatant liquor obtain water-soluble dietary fibre through concentrated, alcohol precipitation; 4. after step, 1. middle filtration gained filter residue washes with water, by liquid ratio, 3:1～8:1 adds water, successively with α-amylase insulation liquefaction and saccharifying enzyme insulation saccharification, centrifugation, must precipitate and supernatant liquor, by precipitation wash with water, lyophilize obtains water insoluble dietary fiber; Described α-amylase insulation liquefaction top condition is pH value 4～6, α-amylase addition 0.05～0.15%, 60～80 ℃ of liquefaction temperatures, liquefying time 50～70 min, and saccharifying enzyme insulation saccharification top condition is pH value 3～4, saccharifying enzyme addition 0.05～0.15%, 55～65 ℃ of saccharification temperatures, saccharification time 10～20 min; 5. 4. active dry yeast constant temperature oscillation fermentation for centrifugation gained supernatant liquor of step, the gained fermented liquid obtains alcohol fuel after distillation, desiccant dehydration.
2. a kind of method of continuous production Cucurmosin, alcohol fuel and food fibre according to claim 1, it is characterized in that the damping fluid that during described step 2., adjusting pH value is used to isoelectric points of proteins is 0.1mol/L acetic acid-sodium acetate solution, the Cucurmosin isoelectric pH value is in 3 ~ 6 scopes.
3. a kind of method of continuous production Cucurmosin, alcohol fuel and food fibre according to claim 1, is characterized in that during step 2. that the centrifugation rotating speed is 5000～10000r/min, in centrifugation time 10～40min scope.
4. a kind of method of continuous production Cucurmosin, alcohol fuel and food fibre according to claim 1, is characterized in that during step 2. that washing precipitation adopts acetone and 95% ethanol continuous washing.
5. a kind of method of continuous production Cucurmosin, alcohol fuel and food fibre according to claim 1, is characterized in that the condition that supernatant liquor during step 3. prepares water-soluble dietary fibre is pH value 6～8,2～6 times of supernatant liquor volumes of dehydrated alcohol consumption, alcohol precipitation times 30～70 min.
6. a kind of method of continuous production Cucurmosin, alcohol fuel and food fibre according to claim 1, is characterized in that during step 5. that supernatant liquor is pH value 4～5, inoculum of dry yeast 0.05～0.10%, 28～30 ℃ of leavening temperatures, fermentation time 65～75 h by the active dry yeast constant temperature oscillation top condition of fermenting.
7. a kind of method of continuous production Cucurmosin, alcohol fuel and food fibre according to claim 1, while it is characterized in that during step 5. fermented liquid is prepared into to alcohol fuel, the dehydration siccative that uses is unslaked lime.
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| CN1830486A (en) * | 2004-11-05 | 2006-09-13 | 福建医科大学 | Application of pumpkin protein in preparation of medicine |
| CN101195835A (en) * | 2007-12-07 | 2008-06-11 | 李传生 | Production technique for fermenting ethyl alcohol with squash as raw material |
| CN101455240A (en) * | 2008-12-29 | 2009-06-17 | 东北农业大学 | Pumpkin seed oil extraction method using water enzyme method |
| CN102180991A (en) * | 2011-04-06 | 2011-09-14 | 武汉普赛特膜技术循环利用有限公司 | Comprehensive utilization method of pumpkin |
| CN102524698A (en) * | 2012-01-19 | 2012-07-04 | 中国计量学院 | Method for preparing pumpkin dietary fiber |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1830486A (en) * | 2004-11-05 | 2006-09-13 | 福建医科大学 | Application of pumpkin protein in preparation of medicine |
| CN101195835A (en) * | 2007-12-07 | 2008-06-11 | 李传生 | Production technique for fermenting ethyl alcohol with squash as raw material |
| CN101455240A (en) * | 2008-12-29 | 2009-06-17 | 东北农业大学 | Pumpkin seed oil extraction method using water enzyme method |
| CN102180991A (en) * | 2011-04-06 | 2011-09-14 | 武汉普赛特膜技术循环利用有限公司 | Comprehensive utilization method of pumpkin |
| CN102524698A (en) * | 2012-01-19 | 2012-07-04 | 中国计量学院 | Method for preparing pumpkin dietary fiber |
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| 胡萝卜渣中膳食纤维提取工艺研究;邵焕霞;《食品与发酵科技》;20090825;第45卷(第04期);摘要、第55页 引言、第56页 1.3 实验方法 * |
| 邵焕霞.胡萝卜渣中膳食纤维提取工艺研究.《食品与发酵科技》.2009,第45卷(第04期),摘要、第55页 引言、第56页 1.3 实验方法. |
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