CN102443607B - Method for preparing fuel alcohol by fermentation - Google Patents
Method for preparing fuel alcohol by fermentation Download PDFInfo
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- CN102443607B CN102443607B CN201110322900.4A CN201110322900A CN102443607B CN 102443607 B CN102443607 B CN 102443607B CN 201110322900 A CN201110322900 A CN 201110322900A CN 102443607 B CN102443607 B CN 102443607B
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Abstract
The invention discloses a method for preparing fuel alcohol by fermentation. The method comprises the following steps: 1) liquidizing: mixing cassava flour and water, boiling at the temperature of 85 DEG C-90 DEG C for 55-65 minutes, then adding amylase, carrying out enzymatic hydrolysis at the temperature of 80-85 DEG C for 55-65 minutes to obtain liquefied wort; wherein the addition of the amylase is 8-12U/g of cassava flour; and the mass ratio of the cassava flour to water is 1: (2.0-2.3); 2) saccharizing: adjusting the pH value of the liquefied wort to 4.0-4.5, adding glucoamylase, carrying out enzymatic hydrolysis at the temperature of 60-65 DEG C for 5-15 minutes to obtain sweet mash; wherein the addition of glucoamylase is 140-160U/g of cassava flour. According to the invention, the yield for preparing fuel ethanol is high, the starch utilization rate is high. The method of the invention uses cassava as a raw material, and has the beneficial effects of wide material source, low production cost and saved energy. The prepared fuel ethanol is used for automobile fuel and is capable of alleviating the problems of energy supply and demand. Therefore, the method has an important significance.
Description
Technical field
The present invention relates to a kind of method of preparing fuel alcohol by fermentation.
Background technology
Fuel alcohol more and more receives people's concern in recent years.Because fuel alcohol has been alleviated the problem of petroleum-based energy shortage as bioenergy, use fuel alcohol can alleviate that temperature that global greenhouse effect aggravation brings raises and serious consequence and the problem of environmental pollution such as terrestrial climate deterioration.But shortage is prepared the method for fuel alcohol efficiently at present.
Cassava (cassava) is perennial plant, and the knob of Perenniporia martius is grain and the economic tuber crops of plantation extensively, and planting area is mainly distributed in the provinces such as Guangxi, Guangdong, Yunnan, Hainan and Fujian.Due to its strong adaptability, water-fast, drought-enduring, impoverishment tolerant, in various soil, as deserted mountain, barren hill can be grown, therefore cassava planting area is very wide, output is also very large.
Summary of the invention
An object of the present invention is to provide a kind of method of preparing fuel alcohol by fermentation.
The method of preparing fuel alcohol by fermentation provided by the present invention, comprises the steps:
1) liquefaction: Tapioca Starch is mixed with water, first 85 ℃～90 ℃ boiling 55～65min, then add amylase, 80～85 ℃ of enzymolysis 55～65min, obtain liquefied fermented glutinous rice; Diastatic addition is 8～12U/g Tapioca Starch; The mass ratio of described Tapioca Starch and water is 1: (2.0～2.3);
2) saccharification: liquefied fermented glutinous rice is adjusted to pH to 4.0～4.5, add saccharifying enzyme, 60～65 ℃ of enzymolysis 5～15min, obtain converted mash; The addition of saccharifying enzyme is 140～160U/g Tapioca Starch;
3) fermentation: add nitrogenous source, sylvite, magnesium salts and yeast in described converted mash, cultivate for the first time 1.5～2.5 hours for 28～32 ℃; At 28～32 ℃, cultivate for the second time 70～74 hours again, obtain fuel alcohol; In the nitrogenous source adding, the amount of N element is 0.6g/L～1.0g/L converted mash, and the amount of the Potassium In Potassium element of interpolation is 0.4g/L～0.6g/L converted mash, and in the magnesium salts of interpolation, the amount of magnesium elements is 0.1g/L～0.3g/L converted mash, and the saccharomycetic amount of interpolation is 1.2 * 10
6～1.2 * 10
8individual yeast/ml converted mash.
In aforesaid method, described step 1) in, described amylase is α-amylase.
In aforesaid method, described step 3) in, before described interpolation N source, sylvite, magnesium salts and yeast, comprise and the temperature of described converted mash is down to the step of 28～32 ℃.
In aforesaid method, described step 3) in, the described step comprising with the velocity fluctuation of 140～180 revs/min of cultivating for the first time.
In aforesaid method, described cassava is that starch content is more than 75% cassava (concrete kind is south China 205); Described nitrogenous source is urea; Described sylvite is KH
2pO
4; Described magnesium salts is MgSO
4; Described yeast is Angel Yeast.
The yield that technique of the present invention is prepared alcohol fuel is high, and starch utilization ratio is high.The inventive method is used cassava as raw material, has that material source is extensive, production cost is low, the beneficial effect of save energy.The alcohol fuel that the present invention is made, for automobile fuel, is alleviated the contradiction of energy supply and demand.Therefore, technique of the present invention is significant.
Accompanying drawing explanation
Fig. 1 is the impact of solid-liquid ratio on zymamsis.
Fig. 2 is the impact of cooking and liquefaction temperature on ethanol production.
Fig. 3 is the impact of α-amylase consumption on ethanol production.
Fig. 4 is the impact of saccharification temperature on ethanol production.
Fig. 5 is the impact of saccharifying enzyme consumption on ethanol production.
Fig. 6 is the impact of inoculum size on ethanol production.
Fig. 7 is the impact of duration of oscillation on ethanol production.
Fig. 8 is the impact of leavening temperature on ethanol production
Fig. 9 is the impact of amount of urea on ethanol production
Figure 10 is cassava performing thick mash alcohol fermentation kinetic curve
Embodiment
The experimental technique using in following embodiment if no special instructions, is ordinary method.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
Use that commercially available cassava is dry or self-control cassava is dry (cassava of fresh results is washed with flowing water, remove the impurity such as mudstone, then with the manual epidermis that goes of the instruments such as pocket knife, again by piece undercut flakiness, be placed in dry 2 days of the loft drier of 50-55 ℃ or utilize sunlight to be exposed to the sun 3 days to dry (determining to be exposed to the sun the time depending on local weather condition), grinding and sieving, is stored in plastic containers standby).
Groping of embodiment 1, processing condition
One, solid-liquid ratio determines
Other technological condition for fermentation are set as, and conditions of cooking is 85～90 ℃, 90min, and liquefaction condition is that α-amylase consumption is 20U/g, 85～90 ℃ of liquefaction 90min.Saccharification pH is 4.0～4.5, and saccharifying enzyme consumption is at 200U/g, 60～65 ℃ of saccharification 30min, and the initial pH that ferments is 4.0～4.5, and inoculum size is 0.2% (mL/mL), and ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.05% (g/mL), KH
2pO
4be 0.02% (g/mL), 28 ℃, 160 revs/min of shaking culture casees are cultivated 1 hour, then 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as Fig. 1.Research shows, solid-liquid ratio was at 1: 3.0 o'clock, its alcohol number of degrees are not high, and residual reducing sugar and residual total reducing sugar are all very little, and at 1: 2.0 o'clock, the alcohol number of degrees are more than 15%, but residual sugar content is higher, in the research of 3～4%, Er Li will army etc., at solid-liquid ratio, it is 1: 2.1 o'clock, wine part is 15.8% (v/v), and residual total reducing sugar reaches 1.84% (g/mL).Consider the potentiality that alcohol output improves, choose the solid-liquid ratio of 1: 2.0 and carry out other tests.
Two, the impact of liquefaction temperature on ethanol production
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0 adopts differing temps boiling 90min, and liquefaction condition is that α-amylase consumption is 20U/g, liquefaction 90min.Saccharification pH is 4.0～4.5, and saccharifying enzyme consumption is at 200U/g, 60～65 ℃ of saccharification 30min, and the initial pH that ferments is 4.0～4.5, and inoculum size is 0.2% (mL/mL), and ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL), 28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as Fig. 2. and test and show, liquefaction temperature reaches in the time of 80～85 ℃, and the impact of temperature is just smaller.
Three, the impact of α-amylase consumption on ethanol production
Other technological condition for fermentation are that conditions of cooking is 80～85 ℃, 90min, the α-amylase of the different amounts of interpolation, liquefaction 90min.Saccharification pH is 4.0～4.5, and saccharifying enzyme consumption is at 200U/g, 60～65 ℃ of saccharification 30min, and the initial pH that ferments is 4.0～4.5, and inoculum size is 0.2% (mL/mL), and ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL), 28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as Fig. 3. and test and show, α-amylase consumption, when 10u/g, just can be obtained good liquefaction effect.
Four, liquefaction process determines
Other technological condition for fermentation are that the consumption of boiling temperature, cooking time, α-amylase, these four variablees of enzyme liquefaction time are done orthogonal test.Other conditions are that saccharification pH is 4.0～4.5, and saccharifying enzyme consumption is at 200U/g, 60～65 ℃ of saccharification 30min, and the initial pH that ferments is 4.0～4.5, and inoculum size is 0.2% (mL/mL), and ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL), 28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as table 1 and table 2.
Table 1, L16 (45) orthogonal test factor and level
Table 2, cooking and liquefaction orthogonal test
Test shows, in liquefaction condition, the importance of influence factor is enzymolysis time by leading inferior successively, liquefaction temperature, and α-amylase consumption, cooking time, best of breed is A
3b
4c
3d
4or be A
3b
4c
4d
4.
Because cooking time is when 60min is above, less on the impact of zymamsis, in order to reduce energy consumption, final Selection and Constitute A
3b
4c
3d
4.
Because of under optimum combination condition, its ethanol production only has increase slightly, and in order to reduce costs, final liquefaction condition is: solid-liquid ratio 1: 2.0, and 85～90 ℃ of boiling 60min, α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.
Five, the impact of saccharification temperature on ethanol production
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and saccharifying enzyme consumption, at 200U/g, adopts different temperature, saccharification 30min, and the initial pH that ferments is 4.0～4.5, and inoculum size is 0.2% (mL/mL), and ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL), 28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as Fig. 4. and test and show, when liquefaction temperature reaches 55～60 ℃, the impact of temperature is just smaller, and ethanol production is all more than 15% (v/v).
Six, the impact of saccharifying enzyme consumption on ethanol production
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 20U/g, 85～90 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, adds the saccharifying enzyme of different amounts, 55～60 ℃ of temperature, and saccharification 30min, the initial pH that ferments is 4.0～4.5, inoculum size 0.2% (mL/mL), ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL), 28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as Fig. 5. and test and show, saccharifying enzyme consumption, when 100u/g, just can be obtained good saccharification result.
Seven, Mashing process is determined
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification temperature, saccharifying enzyme consumption, these three variablees of saccharification time are done to orthogonal test, and the initial pH that ferments is 4.0～4.5, inoculum size 0.2% (mL/mL), and ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL), 28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as table 3 and table 4.
Table 3, L
9(3
4) orthogonal test factor and level
Table 4, saccharification condition orthogonal test
Test shows, in saccharification condition, the importance of influence factor is saccharification temperature by leading inferior successively, and saccharifying enzyme consumption and sugar are taken time, and best of breed is A
1b
2c
1, A
2b
2c
1or A
3b
2c
1.By checking, find, under higher saccharification temperature, ethanol production increases to some extent, therefore select A
3b
2c
1.
In order to reduce costs, reduced in test the consumption of saccharifying enzyme, though A
3b
2c
1output slightly high, but enzyme dosage is larger, therefore final saccharification condition is defined as: saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, saccharification 10min.
Eight, the impact of inoculum size on ethanol production
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, and saccharification 10min, the initial pH that ferments is 4.0～4.5, adopts different inoculum sizes, ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL).28 ℃, 160 revs/min of shaking culture casees were cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as Fig. 6. and test and show, inoculum size is in 1% (mL/mL), and last alcohol output is reasonable, therefore will adopt the inoculum size of 1% (mL/mL).
Nine, the impact of duration of oscillation on ethanol production
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, and saccharification 10min, the initial pH that ferments is 4.0～4.5, inoculum size 1% (mL/mL), ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL).28 ℃, 160 revs/min of shaking culture casees are cultivated the different times, then 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result as shown in Figure 7.Test shows, after earthquake, ethanol production obviously increases, and illustrated that vibration is necessary, and when vibration is in the time of 2 hours, ethanol production is just in a more stable value, therefore this research will be taked to vibrate 2 hours, and then ferments 72 hours.
Ten, the impact of leavening temperature on ethanol production
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, and saccharification 10min, the initial pH that ferments is 4.0～4.5, inoculum size 1% (mL/mL), ammonium sulfate consumption is 0.1% (g/mL), MgSO
4be 0.1% (g/mL), KH
2pO
4be 0.1% (g/mL).28 ℃, 160 revs/min of shaking culture casees were cultivated 2 hours, condition of different temperatures bottom fermentation 72 hours.
Result is as Fig. 8. and experimental study shows, at 28 ± 1 ℃, in the time of 30 ± 1 ℃ of left and right, zymamsis is preferably, along with the rising of temperature, slightly declines.This may be because the cause that temperature raises the murder by poisoning of yeast is increased.Consider the reduction of the cooling energy consumption of actual production, therefore test will adopt 30 ± 1 ℃.
11, N source and the impact of other inorganic salt on ethanol production
(1) N source and other inorganic salt orthogonal tests
Other technological condition for fermentation are, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, and saccharification 10min, the initial pH that ferments is 4.0～4.5, inoculum size is 1% (mL/mL).To ammonium sulfate consumption, MgSO
4consumption, KH
2pO
4consumption, CaCl
2these four factors of consumption were done orthogonal test, and 28 ℃, 160 revs/min of shaking culture casees are cultivated 1 hour, 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result is as table 5 and table 6.
Table 5, L16 (45) orthogonal test factor and level
Table 6, N source and the impact of other inorganic salt on ethanol production
Test shows, the importance of influence factor is nitrogenous source by leading inferior successively, KH
2pO
4, calcium ion, magnesium ion, best of breed is A
1b
3c
1d
2or A
1b
4c
1d
2.The final A that selects
1b
3c
1d
2, urea 0.1% (g/mL), potassium primary phosphate 0.02% (g/mL), magnesium sulfate 0.05% (g/mL), without extra interpolation calcium ion.
(2) amount of urea determines
Research shows, when the consumption of urea reaches 0.08% (g/mL), the productive rate of alcohol just can be in a higher level, and residual total reducing sugar and residual reducing sugar also lower, so select 0.08% (g/mL) as the dosage adding, fermentation is now just more stable, the alcohol number of degrees are at 16.2%～17.5% (v/v), residual sugar is at 2%～4% (g/mL), the more initial fermentation results of alcoholic strength has had significant raising, residual sugar is greatly reduced, but is still in a higher level.
12, cassava performing thick mash alcohol fermentation kinetic curve
Technological condition for fermentation is, the solid-liquid ratio of 1: 2.0, and 85～90 ℃ of boiling 60min, liquefaction condition is that α-amylase consumption is 20U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, saccharification 10min, the initial pH that ferments is 4.0～4.5, inoculum size is 1%, add urea 0.08% (g/mL), potassium primary phosphate 0.02% (g/mL), magnesium sulfate 0.05% (g/mL).28 ℃, 160 revs/min of shaking culture casees are cultivated 2 hours, then 30 ± 1 ℃ of condition bottom fermentations 72 hours.
Result as shown in figure 10.Research shows, after fermentation to 72 hour, the weightlessness of fermentation system is just fewer, alcoholic strength also increase seldom, total reducing sugar and reducing sugar are also reduced to a lower level, consider the utilization ratio of equipment, and shorten fermentation period as far as possible, therefore the fermentation time of 72 hours is more rational.
13, the further optimization of technique
By the solid-liquid ratio test of 1: 2.0, we drew such conclusion: cassava performing thick mash alcohol fermentation, and the alcohol number of degrees are at 16.2%～17.5% (v/v), and residual total reducing sugar is 2%～3%, and residual reducing sugar is in 1% left and right, and residual sugar is higher, needs to improve.When guaranteeing alcohol in high yield, improve raw material availability, this research is started with from reduction solid-liquid ratio.Other technological condition for fermentation are, 85～90 ℃ of boiling 60min, and liquefaction condition is that α-amylase consumption is 10U/g, 80～85 ℃ of liquefaction 60min.Saccharification pH is 4.0～4.5, and the consumption of saccharifying enzyme is 150U/g, 60～65 ℃ of temperature, saccharification 10min, the initial pH that ferments is 4.0～4.5, inoculum size is 1%, add urea 0.08% (g/mL), potassium primary phosphate 0.02% (g/mL), magnesium sulfate 0.05% (g/mL).28 ℃, 160 revs/min of shaking culture casees are cultivated 2 hours, then 30 ± 1 ℃ of condition bottom fermentations 72 hours.Test-results is as following table 7:
Table 17 under optimum process condition different feed liquid than the comparison of Tapioca Starch thick mash fermentation result
From producing actual consideration, the solid-liquid ratio of 1: 2.3, mash viscosity is minimum, and total reducing sugar and reducing sugar all lower, on producing, be the most feasible.1: 2.1 and 1: although 2.0 the solid-liquid ratio alcohol number of degrees compared with 1: 2.3 to exceed 1 about degree, residual total reducing sugar will exceed respectively 1% and 2% left and right, this is obviously uneconomic.
The processing step of the inventive method is as follows:
Raw materials pretreatment: the cassava of fresh results is washed with flowing water, remove the impurity such as mudstone, then with the manual epidermis that goes of the instruments such as pocket knife, again by piece undercut flakiness, be placed in dry 2 days of the loft drier of 50～55 ℃ or utilize sunlight to be exposed to the sun 3 days to dry (determining to be exposed to the sun the time depending on local weather condition), after pulverizing, cross 40 mesh sieves, be stored in plastic containers standby.
The preparation of bacterium liquid: be the aqueous solution of the white sugar of 5% (quality percentage composition) by 1g yeast and 10mL concentration, after 30 minutes, all substances in culture vessel are bacterium liquid 37 ℃ of activation, and in bacterium liquid, saccharomycetic concentration is at least 1.2 * 10
8individual/mL.
Liquefaction: Tapioca Starch is mixed with water, first 85 ℃～90 ℃ boiling 55～65min, then add amylase, 80～85 ℃ of enzymolysis 55～65min, obtain liquefied fermented glutinous rice; Diastatic addition is 8～12U/g Tapioca Starch; The mass ratio of described Tapioca Starch and water is 1: (2.0～2.3);
Saccharification: liquefied fermented glutinous rice is adjusted to pH to 4.0～4.5, add saccharifying enzyme, 60～65 ℃ of enzymolysis 5～15min, obtain converted mash; The addition of saccharifying enzyme is 140～160U/g Tapioca Starch;
Fermentation: the temperature of described converted mash is down to 28～32 ℃, adds nitrogenous source, sylvite, magnesium salts and yeast in described converted mash, cultivate 1.5～2.5 hours with the velocity fluctuation of 160 revs/min for the first time for 28～32 ℃; At 28～32 ℃, cultivate for the second time 70～74 hours again, obtain fuel alcohol; In the nitrogenous source adding, the amount of N element is 0.6g/L～1.0g/L converted mash, and the amount of the Potassium In Potassium element of interpolation is 0.4g/L～0.6g/L converted mash, and in the magnesium salts of interpolation, the amount of magnesium elements is 0.1g/L～0.3g/L converted mash, and the saccharomycetic amount of interpolation is 1.2 * 10
6～1.2 * 10
8individual yeast/ml converted mash.
The raw material using in method is as follows:
Cassava Cultivars is south China 205, and wherein starch content is 79.74%.
Yeast is Angel Yeast.
Amylase is α-amylase; Name of product is Suhong AA Plus, purchased from Enzyme Co., Ltd processed of Suzhou HTC; 35000U/mL.
Saccharifying enzyme is α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme; Name of product is Suhong GA475; Purchased from Enzyme Co., Ltd processed of Suzhou HTC; 100000U/mL.
Nitrogenous source is urea; Sylvite is KH
2pO
4; Magnesium salts is MgSO
4.
In this technique, the detection method of karusen ethanol content: GB/T 394.2-2008.
With accurate Ebullioscope, read alcohol volume fraction indicating value, by appendix A, carry out temperature correction, try to achieve the volume fraction of ethanol content in the time of 20 ℃, be alcoholic strength.
In this technique, the detection method of starch utilization ratio: acid-hydrolysis method in GB/T 5009.9-2008.
Method I:
Liquefaction: Tapioca Starch is mixed with water, first 88 ℃ of boiling 60min, then add amylase, 83 ℃ of enzymolysis 60min, obtain liquefied fermented glutinous rice; Diastatic addition is 10U/g Tapioca Starch; The mass ratio of described Tapioca Starch and water is 1: 2.3;
Saccharification: liquefied fermented glutinous rice is adjusted to pH to 4.3, add saccharifying enzyme, 63 ℃ of enzymolysis 10min, obtain converted mash; The addition of saccharifying enzyme is 150U/g Tapioca Starch;
Fermentation: the temperature of described converted mash is down to 30 ℃, adds nitrogenous source, sylvite, magnesium salts and yeast in described converted mash, cultivate 2 hours with the velocity fluctuation of 160 revs/min for the first time for 30 ℃; At 30 ℃, cultivate for the second time 72 hours again, obtain fuel alcohol; In the nitrogenous source adding, the amount of N element is 0.8g/L converted mash, and the amount of the Potassium In Potassium element of interpolation is 0.5g/L converted mash, and in the magnesium salts of interpolation, the amount of magnesium elements is 0.2g/L converted mash, and the saccharomycetic amount of interpolation is 1.2 * 10
8individual yeast/ml converted mash.
Method II:
1) liquefaction: Tapioca Starch is mixed with water, first 85 ℃ of boiling 55min, then add amylase, 80 ℃ of enzymolysis 55min, obtain liquefied fermented glutinous rice; Diastatic addition is 8U/g Tapioca Starch; The mass ratio of described Tapioca Starch and water is 1: 2.0;
2) saccharification: liquefied fermented glutinous rice is adjusted to pH to 4.0, add saccharifying enzyme, 60 ℃ of enzymolysis 5min, obtain converted mash; The addition of saccharifying enzyme is 140U/g Tapioca Starch;
3) fermentation: the temperature of described converted mash is down to 28 ℃, adds nitrogenous source, sylvite, magnesium salts and yeast in described converted mash, cultivate 1.5 hours with the velocity fluctuation of 140 revs/min for the first time for 28 ℃; At 28 ℃, cultivate for the second time 70 hours again, obtain fuel alcohol; In the nitrogenous source adding, the amount of N element is 0.6g/L converted mash, and the amount of the Potassium In Potassium element of interpolation is 0.4g/L converted mash, and in the magnesium salts of interpolation, the amount of magnesium elements is 0.1g/L converted mash, and the saccharomycetic amount of interpolation is 1.2 * 10
6individual yeast/ml converted mash.
Method III:
1) liquefaction: Tapioca Starch is mixed with water, first 90 ℃ of boiling 65min, then add amylase, 85 ℃ of enzymolysis 65min, obtain liquefied fermented glutinous rice; Diastatic addition is 12U/g Tapioca Starch; The mass ratio of described Tapioca Starch and water is 1: 2.1;
2) saccharification: liquefied fermented glutinous rice is adjusted to pH to 4.5, add saccharifying enzyme, 65 ℃ of enzymolysis 15min, obtain converted mash; The addition of saccharifying enzyme is 160U/g Tapioca Starch;
3) fermentation: add nitrogenous source, sylvite, magnesium salts and yeast in described converted mash, cultivate for the first time 2.5 hours for 32 ℃; At 32 ℃, cultivate for the second time 74 hours again, obtain fuel alcohol; In the nitrogenous source adding, the amount of N element is 1.0g/L converted mash, and the amount of the Potassium In Potassium element of interpolation is 0.6g/L converted mash, and in the magnesium salts of interpolation, the amount of magnesium elements is 0.3g/L converted mash, and the saccharomycetic amount of interpolation is 1.2 * 10
7individual yeast/ml converted mash.
Result:
Method I: karusen ethanol content is 16.3% (v/v); Starch utilization ratio is 93.7%.
Method II: karusen ethanol content is 17.0% (v/v); Starch utilization ratio is 88.9%.
Method III: karusen ethanol content is 17.0% (v/v); Starch utilization ratio is 91.8%.
Claims (4)
1. a method for preparing fuel alcohol by fermentation, comprises the steps:
1) liquefaction: Tapioca Starch is mixed with water, first 85 ℃～90 ℃ boiling 55 ~ 65min, then add amylase, 80～85 ℃ of enzymolysis 55 ~ 65min, obtain liquefied fermented glutinous rice; Diastatic addition is 8 ~ 12 U/g Tapioca Starchs; The mass ratio of described Tapioca Starch and water is 1:(2.0 ~ 2.3);
2) saccharification: liquefied fermented glutinous rice is adjusted to pH to 4.0～4.5, add saccharifying enzyme, 60～65 ℃ of enzymolysis 5 ~ 15 min, obtain converted mash; The addition of saccharifying enzyme is 140 ~ 160U/g Tapioca Starch;
3) fermentation: add nitrogenous source, sylvite, magnesium salts and yeast in described converted mash, cultivate for the first time 1.5 ~ 2.5 hours for 28 ~ 32 ℃; At 28 ~ 32 ℃, cultivate for the second time 70 ~ 74 hours again, obtain fuel alcohol; In the nitrogenous source adding, the amount of N element is 0.6g/L ~ 1.0g/L converted mash, and the amount of the Potassium In Potassium element of interpolation is 0.4g/L ~ 0.6g/L converted mash, and in the magnesium salts of interpolation, the amount of magnesium elements is 0.1g/L ~ 0.3g/L converted mash, and the saccharomycetic amount of interpolation is 1.2 * 10
6~ 1.2 * 10
7individual yeast/ml converted mash;
In described step 3), the described step comprise with the velocity fluctuation of 140 ~ 180 revs/min of cultivating for the first time.
2. method according to claim 1, is characterized in that: in described step 1), described amylase is α-amylase.
3. method according to claim 1 and 2, is characterized in that: in described step 3), before described interpolation N source, sylvite, magnesium salts and yeast, comprise and the temperature of described converted mash is down to the step of 28 ~ 32 ℃.
4. method according to claim 1 and 2, is characterized in that: described cassava is that starch content is more than 75% cassava; Described nitrogenous source is urea; Described sylvite is KH
2pO
4; Described magnesium salts is MgSO
4; Described yeast is Angel Yeast.
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| CN110499342A (en) * | 2018-05-16 | 2019-11-26 | 南京百斯杰生物工程有限公司 | Application of the non-starch class polysaccharase in cereal deep processing |
| CN111562360B (en) * | 2020-04-10 | 2024-04-16 | 深圳市安途锐科技有限公司 | Temperature compensation alcohol detection method and alcohol detection instrument |
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| 唐艳艳等.木薯粉浓醪酒精同步糖化发酵工艺研究.《安徽农业科学》.2010,第38卷(第23期),12690－12692. |
| 木薯粉浓醪酒精同步糖化发酵工艺研究;唐艳艳等;《安徽农业科学》;20100831;第38卷(第23期);12690－12692 * |
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