CN105063110B - A kind of method using bagasse production lactic acid - Google Patents
A kind of method using bagasse production lactic acid Download PDFInfo
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Abstract
A kind of method using bagasse production lactic acid of the present invention, first, to after the pre-processing of bagasse, it is 4.8 that addition distilled water, which adjusts pH value, then adds zytase and cellulase, is put on bed and carries out enzymatic saccharification；Filtered after saccharification, obtain filtering enzymolysis liquid；Obtained filtering enzymolysis liquid is concentrated, obtains concentration enzymolysis liquid；Filtered after saccharification, obtain filtering enzymolysis liquid；Obtained filtering enzymolysis liquid is concentrated, obtains concentration enzymolysis liquid；Take the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, be then placed in shaking table culture fermentation；Ferment after 24h, take out, be inoculated with the seed liquor of bacillus coagulans；It is subsequently placed in shaking table culture fermentation；Continue the 10h that ferments, the seed liquor of inoculated aspergillus niger is fermented；After resulting zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.The content of Lactic Acid from Fermentation Broth is high obtained by the method for the present invention, and purity is high.
Description
Technical field
The present invention relates to a kind of method for producing lactic acid, specifically a kind of method using bagasse production lactic acid.
Background technology
Lactic acid is a kind of important organic acid, has a wide range of applications in recent years in industries such as chemical industry, food, medicine,
The exploitation of polylactic acid-based biodegradable plastic makes the demand of lactic acid surge.Its unique structure advantage that lactic acid has with it is shown
It is widely applied prospect.The significantly increasing of exhaustion and Novel environment-friendlymaterial material with petroleum resources --- polylactic acid demand
Add, people are striving to find the new raw material for substituting oil-fired new energy and reducing L. production of lactic acid costs；
Sugarcane is one of primary raw material of sugaring.The remaining bagasse after squeezing sugar, there are about 50% fiber can use
Carry out papermaking.But, wherein still there is part sugarcane marrow (myelocyte) there is no intertexture power, should be removed before pulping process.Bagasse fibre
Length is about 0.65-2.17mm, and width is 21-28 μm.Although its fibre morphology is less than timber and bamboo, compare paddy and wheat
Grass fiber is then slightly better.After slurry can be incorporated part wood pulp, offset printing paper processed, cement bag paper etc. are copied.
The content of the invention
The present invention seeks to the deficiency for solution above-mentioned technical problem, there is provided a kind of side using bagasse production lactic acid
Method, the content of Lactic Acid from Fermentation Broth is high, after testing up to 198g/L-205g/L.
The present invention is that technical solution is used by solving above-mentioned technical problem：
A kind of method using bagasse production lactic acid, it is characterised in that：Comprise the following steps：
Step 1: the pretreatment of bagasse:Bagasse powder is broken to 100-200 mesh, it is spare；It is 0.5% with mass fraction
NaOH solution sprinkling rubbing, time 12h, bagasse after being pre-processed are carried out to the bagasse after crushing；Operating method
For：Rubbing 5min, is put into 45 DEG C of environment after sprinkling uniformly；NaOH solution is sprayed every 4h and is rubbed once, every time rubbing
5min；
Step 2: it is 4.8 to add distilled water in bagasse after the pre-treatment to adjust pH value, zytase and fibre are then added
The plain enzyme of dimension, is put on 50 DEG C of shaking tables and carries out enzymatic saccharification, saccharificatinn period 8h-10h；The zytase and cellulase add
Enter amount is respectively bagasse quality after pre-processing 0.05% and 0.0 8%,
Step 3: being filtered after saccharification, filtering enzymolysis liquid is obtained；Obtained filtering enzymolysis liquid is concentrated,
Obtain concentration enzymolysis liquid；
Cultivated Step 4: taking Lactobacillus casei to be inoculated in Lactobacillus casei seed liquid culture medium, obtain cheese breast
The seed liquor of bacillus, it is spare；Take bacillus coagulans to be inoculated in bacillus coagulans seed liquid culture medium to be cultivated, coagulated
The seed liquor of bacillus is tied, it is spare；Aspergillus niger is taken, is inoculated in aspergillus niger seed liquid culture medium and is cultivated, obtain black song
Mould seed liquor；
Step 5: taking the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, inoculum concentration is fermentation medium volume
2-2.3%；Then, shaking table culture fermentation, 35 DEG C of temperature, speed 150r/min are placed in；Ferment after 24h, take out, inoculation is solidifying
The seed liquor of bacillus is tied, inoculum concentration is the 1.5-2% of fermentation medium；After continuing fermentation 10h, take out, inoculated aspergillus niger
Seed liquor, inoculum concentration are the 1.5-2% of fermentation medium；It is subsequently placed in shaking table culture fermentation, 32 DEG C of temperature, speed 200r/
Min, fermentation time 55h-65h；After resulting zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.
The method that is concentrated of filtering enzymolysis liquid is：Under 0.09 atmospheric pressure, bath temperature reaches 53 DEG C
There is boiling phenomenon, rotating speed 60-80r/min in saccharified liquid.
The seed culture medium of the Lactobacillus casei, 50g containing glucose in every liter of culture medium, 10 g of peptone, beef extract
5 g, 5 g of yeast extract, anhydrous sodium acetate 5 g, MgSO4.7H2O 20mL, Mn SO4.7H2O 20mL, calcium carbonate 20g, pH
6.8 and tween 1mL, surplus are water.
The seed culture culture medium of the bacillus coagulans, 80 g containing glucose in every liter of culture medium, (NH)4SO4 4
G, KH2PO40.3 g, ZnSO4.7H2O 0.44 g, MgSO4.7H20.25 g of O, surplus are water.
The fermentation medium, 50ml containing enzymolysis liquid, peptone 10 g, yeast extract 5g in every liter of culture medium,
MgSO4.7H2O 0. 5 g, (NH)4SO44 g, KH2PO40.4, NaCl 0.1g and CaCO43g, surplus is water.
The preparation method of the aspergillus niger seed liquid culture medium is：Every 1000 milliliters of culture mediums, take 200 grams of potato,
It is cut into small pieces, adds 1000 milliliters of water to boil 30 minutes, removes potato ball with filtered through gauze after cooling, grape is added into filtrate
To being completely dissolved, moisturizing to 1000 milliliters, 121 DEG C sterilizes 20 minutes 20 grams of stirrings of sugar.
The method of the Lactic Acid from Fermentation Broth assay is：Analysis Lactic Acid from Fermentation Broth is carried out using high performance liquid chromatography
Content, chromatographic column are the C18 anti-phase (4.6mm × 250mm, 5 μm) of Di Ma companies；Mobile phase is the phosphoric acid of 0.01mol/L
Two ammonium salt solution of hydrogen, with phosphoric acid tune pH values to 2.6, after 0.45 μm of aperture synthetic cellulose esters membrane filtration, ultrasound degassing 1h；Stream
Fast 0.8mL/min, Detection wavelength 210nm, 20 μ L of sample size, column temperature is room temperature.
Beneficial effect is：
The present invention uses the method that bagasse produces lactic acid, bagasse is pre-processed in this method, fibre material makes
The first step, and a very crucial step.Cellulase is mainly set directly to be contacted with lignocellulosic substrate more thorough,
Reactivity of the cellulase to lignocellulosic is improved, in addition can improve the vigor of cellulase.Contain cellulose in bagasse,
Hemicellulose, lignin.It is difficult that hydrolysis is complete to be individually added into a kind of enzyme, while it is bagasse to add cellulase and zytase
Hydrolysis is more complete.Traditional zymotic enzymatic saccharification liquid only uses a kind of bacterium, and saccharified liquid utilization rate is not high.Lactobacillus casei is mainly sharp
With glucose, and bacillus coagulans mainly utilizes xylose.Bacillus coagulans is about 1 in glucose and xylose ratio:When 3
Bacillus coagulans is to utilize xylose efficiency highest, so glucose and xylose ratio after when addition Lactobacillus casei fermentation 24 is small
About 1:3, add bacillus coagulans, bacillus coagulans efficiency highest.The present invention is former using bagasse production lactic acid
Expect for agricultural byproduct, but the content of Lactic Acid from Fermentation Broth is high, after testing up to 198g/L-205g/L, and purity reach 98% with
On；It is economical and the technique is pollution-free, environmental protection.
Embodiment
A kind of method using bagasse production lactic acid, it is characterised in that：Comprise the following steps：
Step 1: the pretreatment of bagasse:Bagasse powder is broken to 100-200 mesh, it is spare；It is 0.5% with mass fraction
NaOH solution sprinkling rubbing, time 12h, bagasse after being pre-processed are carried out to the bagasse after crushing；Operating method
For：Rubbing 5min, is put into 45 DEG C of environment after sprinkling uniformly；NaOH solution is sprayed every 4h and is rubbed once, every time rubbing
5min；
Step 2: it is 4.8 to add distilled water in bagasse after the pre-treatment to adjust pH value, zytase and fibre are then added
The plain enzyme of dimension, is put on 50 DEG C of shaking tables and carries out enzymatic saccharification, saccharificatinn period 8h-10h；The zytase and cellulase add
Enter amount is respectively bagasse quality after pre-processing 0.05% and 0.0 8%,
Step 3: being filtered after saccharification, filtering enzymolysis liquid is obtained；Obtained filtering enzymolysis liquid is concentrated,
Obtain concentration enzymolysis liquid；
Cultivated Step 4: taking Lactobacillus casei to be inoculated in Lactobacillus casei seed liquid culture medium, obtain cheese breast
The seed liquor of bacillus, it is spare；Take bacillus coagulans to be inoculated in bacillus coagulans seed liquid culture medium to be cultivated, coagulated
The seed liquor of bacillus is tied, it is spare；Aspergillus niger is taken, is inoculated in aspergillus niger seed liquid culture medium and is cultivated, obtain black song
Mould seed liquor；
Step 5: taking the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, inoculum concentration is fermentation medium volume
2-2.3%；Then, shaking table culture fermentation, 35 DEG C of temperature, speed 150r/min are placed in；Ferment after 24h, take out, inoculation is solidifying
The seed liquor of bacillus is tied, inoculum concentration is the 1.5-2% of fermentation medium；After continuing fermentation 10h, take out, inoculated aspergillus niger
Seed liquor, inoculum concentration are the 1.5-2% of fermentation medium；It is subsequently placed in shaking table culture fermentation, 32 DEG C of temperature, speed 200r/
Min, fermentation time 55h-65h；After resulting zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.
The method that is concentrated of filtering enzymolysis liquid is：Under 0.09 atmospheric pressure, bath temperature reaches 53 DEG C
There is boiling phenomenon, rotating speed 60-80r/min in saccharified liquid.
The seed culture medium of the Lactobacillus casei, 50g containing glucose in every liter of culture medium, 10 g of peptone, beef extract
5 g, 5 g of yeast extract, anhydrous sodium acetate 5 g, MgSO4.7H2O 20mL, Mn SO4.7H2O 20mL, calcium carbonate 20g, pH
6.8 and tween 1mL, surplus are water.
The seed culture culture medium of the bacillus coagulans, 80 g containing glucose in every liter of culture medium, (NH)4SO4 4
G, KH2PO40.3 g, ZnSO4.7H2O 0.44 g, MgSO4.7H20.25 g of O, surplus are water.
The preparation method of the aspergillus niger seed liquid culture medium is：Every 1000 milliliters of culture mediums, take 200 grams of potato,
It is cut into small pieces, adds 1000 milliliters of water to boil 30 minutes, removes potato ball with filtered through gauze after cooling, grape is added into filtrate
To being completely dissolved, moisturizing to 1000 milliliters, 121 DEG C sterilizes 20 minutes 20 grams of stirrings of sugar.
The fermentation medium, 50ml containing enzymolysis liquid, peptone 10 g, yeast extract 5g in every liter of culture medium,
MgSO4.7H2O 0. 5 g, (NH)4SO44 g, KH2PO40.4, NaCl 0.1g and CaCO43g, surplus is water.
The method of the Lactic Acid from Fermentation Broth assay is：Analysis Lactic Acid from Fermentation Broth is carried out using high performance liquid chromatography
Content, chromatographic column are the C18 anti-phase (4.6mm × 250mm, 5 μm) of Di Ma companies；Mobile phase is the phosphoric acid of 0.01mol/L
Two ammonium salt solution of hydrogen, with phosphoric acid tune pH values to 2.6, after 0.45 μm of aperture synthetic cellulose esters membrane filtration, ultrasound degassing 1h；Stream
Fast 0.8mL/min, Detection wavelength 210nm, 20 μ L of sample size, column temperature is room temperature.
Embodiment 1
A kind of method using bagasse production lactic acid, it is characterised in that：Comprise the following steps：
Step 1: the pretreatment of bagasse:Bagasse powder is broken to 100 mesh, it is spare；It is 0.5% with mass fraction
NaOH solution carries out the bagasse after crushing sprinkling rubbing, time 12h, bagasse after being pre-processed；Operating method is：
Rubbing 5min, is put into 45 DEG C of environment after sprinkling uniformly；NaOH solution is sprayed every 4h and is rubbed once, rubs 5min every time；
Step 2: it is 4.8 to add distilled water in bagasse after the pre-treatment to adjust pH value, zytase and fibre are then added
The plain enzyme of dimension；It is put on 50 DEG C of shaking tables and carries out enzymatic saccharification, saccharificatinn period 8h-10h；The zytase and cellulase add
Enter amount is respectively bagasse quality after pre-processing 0.05% and 0.0 8%；
Step 3: being filtered after saccharification, filtering enzymolysis liquid is obtained；Obtained filtering enzymolysis liquid is concentrated,
Obtain concentration enzymolysis liquid；Under 0.09 atmospheric pressure, bath temperature reaches 53 DEG C of saccharified liquids and boiling phenomenon, rotating speed occurs
60r/min；
Cultivated Step 4: taking Lactobacillus casei to be inoculated in Lactobacillus casei seed liquid culture medium, obtain cheese breast
The seed liquor of bacillus, it is spare；Take bacillus coagulans to be inoculated in bacillus coagulans seed liquid culture medium to be cultivated, coagulated
The seed liquor of bacillus is tied, it is spare；Aspergillus niger is taken, is inoculated in aspergillus niger seed liquid culture medium and is cultivated, obtain black song
Mould seed liquor；
Step 5: taking the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, inoculum concentration is fermentation medium volume
2%；Then, shaking table culture fermentation, 35 DEG C of temperature, speed 150r/min are placed in；Ferment after 24h, take out, inoculation condenses bud
The seed liquor of spore bacillus, inoculum concentration are the 1.5% of fermentation medium；After continuing fermentation 10h, take out, the seed of inoculated aspergillus niger
Liquid, inoculum concentration are the 1.5% of fermentation medium；It is subsequently placed in shaking table culture fermentation, 32 DEG C, speed 200r/min of temperature, fermentation
Time 55h；After resulting zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.
The fermentation medium, 50ml containing enzymolysis liquid, peptone 10 g, yeast extract 5g in every liter of culture medium,
MgSO4.7H2O 0. 5 g, (NH)4SO44 g, KH2PO40.4, NaCl 0.1g and CaCO43g, surplus is water.
The preparation method of the aspergillus niger seed liquid culture medium is：Every 1000 milliliters of culture mediums, take 200 grams of potato,
It is cut into small pieces, adds 1000 milliliters of water to boil 30 minutes, removes potato ball with filtered through gauze after cooling, grape is added into filtrate
To being completely dissolved, moisturizing to 1000 milliliters, 121 DEG C sterilizes 20 minutes 20 grams of stirrings of sugar.
The method of the Lactic Acid from Fermentation Broth assay is：Analysis Lactic Acid from Fermentation Broth is carried out using high performance liquid chromatography
Content, chromatographic column are the C18 anti-phase (4.6mm × 250mm, 5 μm) of Di Ma companies；Mobile phase is the phosphoric acid of 0.01mol/L
Two ammonium salt solution of hydrogen, with phosphoric acid tune pH values to 2.6, after 0.45 μm of aperture synthetic cellulose esters membrane filtration, ultrasound degassing 1h；Stream
Fast 0.8mL/min, Detection wavelength 210nm, 20 μ L of sample size, column temperature is room temperature, is up to 198g/L, purity in zymotic fluid after testing
Reach 99%.
Embodiment 2
A kind of method using bagasse production lactic acid, it is characterised in that：Comprise the following steps：
Step 1: the pretreatment of bagasse:Bagasse powder is broken to 200 mesh, it is spare；It is 0.5% with mass fraction
NaOH solution carries out the bagasse after crushing sprinkling rubbing, time 12h, bagasse after being pre-processed；Operating method is：
Rubbing 5min, is put into 45 DEG C of environment after sprinkling uniformly；NaOH solution is sprayed every 4h and is rubbed once, rubs 5min every time；
Step 2: it is 4.8 to add distilled water in bagasse after the pre-treatment to adjust pH value, zytase and fibre are then added
The plain enzyme of dimension, is put on 50 DEG C of shaking tables and carries out enzymatic saccharification, saccharificatinn period 10h；The addition of the zytase and cellulase
Amount is respectively 0.05% and 0.0 8% of bagasse quality after pre-processing；
Step 3: being filtered after saccharification, filtering enzymolysis liquid is obtained；Obtained filtering enzymolysis liquid is concentrated,
Obtain concentration enzymolysis liquid；Under 0.09 atmospheric pressure, bath temperature reaches 53 DEG C of saccharified liquids and boiling phenomenon, rotating speed occurs
80r/min；
Cultivated Step 4: taking Lactobacillus casei to be inoculated in Lactobacillus casei seed liquid culture medium, obtain cheese breast
The seed liquor of bacillus, it is spare；Take bacillus coagulans to be inoculated in bacillus coagulans seed liquid culture medium to be cultivated, coagulated
The seed liquor of bacillus is tied, it is spare；Aspergillus niger is taken, is inoculated in aspergillus niger seed liquid culture medium and is cultivated, obtain black song
Mould seed liquor；
Step 5: taking the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, inoculum concentration is fermentation medium volume
2.3%；Then, shaking table culture fermentation, 35 DEG C of temperature, speed 150r/min are placed in；Ferment after 24h, take out, inoculation condenses
The seed liquor of bacillus, inoculum concentration are the 2% of fermentation medium；After continuing fermentation 10h, take out, the seed of inoculated aspergillus niger
Liquid, inoculum concentration are the 2% of fermentation medium；It is subsequently placed in shaking table culture fermentation, 32 DEG C, speed 200r/min of temperature, during fermentation
Between 65h；After resulting zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.
The seed culture medium of the Lactobacillus casei, 50g containing glucose in every liter of culture medium, 10 g of peptone, beef extract
5 g, 5 g of yeast extract, anhydrous sodium acetate 5 g, MgSO4.7H2O 20mL, Mn SO4.7H2O 20mL, calcium carbonate 20g, pH
6.8 and tween 1mL, surplus are water.
The seed culture culture medium of the bacillus coagulans, 80 g containing glucose in every liter of culture medium, (NH)4SO4 4
G, KH2PO40.3 g, ZnSO4.7H2O 0.44 g, MgSO4.7H20.25 g of O, surplus are water.
The preparation method of the aspergillus niger seed liquid culture medium is：Every 1000 milliliters of culture mediums, take 200 grams of potato,
It is cut into small pieces, adds 1000 milliliters of water to boil 30 minutes, removes potato ball with filtered through gauze after cooling, grape is added into filtrate
To being completely dissolved, moisturizing to 1000 milliliters, 121 DEG C sterilizes 20 minutes 20 grams of stirrings of sugar.
The method of the Lactic Acid from Fermentation Broth assay is：Analysis Lactic Acid from Fermentation Broth is carried out using high performance liquid chromatography
Content, chromatographic column are the C18 anti-phase (4.6mm × 250mm, 5 μm) of Di Ma companies；Mobile phase is the phosphoric acid of 0.01mol/L
Two ammonium salt solution of hydrogen, with phosphoric acid tune pH values to 2.6, after 0.45 μm of aperture synthetic cellulose esters membrane filtration, ultrasound degassing 1h；Stream
Fast 0.8mL/min, Detection wavelength 210nm, 20 μ L of sample size, column temperature is room temperature, is up to 201g/L, purity in zymotic fluid after testing
Reach 98%.
Embodiment 3
A kind of method using bagasse production lactic acid, it is characterised in that：Comprise the following steps：
Step 1: the pretreatment of bagasse:Bagasse powder is broken to 150 mesh, it is spare；It is 0.5% with mass fraction
NaOH solution carries out the bagasse after crushing sprinkling rubbing, time 12h, bagasse after being pre-processed；Operating method is：
Rubbing 5min, is put into 45 DEG C of environment after sprinkling uniformly；NaOH solution is sprayed every 4h and is rubbed once, rubs 5min every time；
Step 2: it is 4.8 to add distilled water in bagasse after the pre-treatment to adjust pH value, zytase and fibre are then added
The plain enzyme of dimension；It is put on 50 DEG C of shaking tables and carries out enzymatic saccharification, saccharificatinn period 8h-10h；The zytase and cellulase add
Enter amount is respectively bagasse quality after pre-processing 0.05% and 0.0 8%；
Step 3: being filtered after saccharification, filtering enzymolysis liquid is obtained；Obtained filtering enzymolysis liquid is concentrated,
Obtain concentration enzymolysis liquid；
Cultivated Step 4: taking Lactobacillus casei to be inoculated in Lactobacillus casei seed liquid culture medium, obtain cheese breast
The seed liquor of bacillus, it is spare；Take bacillus coagulans to be inoculated in bacillus coagulans seed liquid culture medium to be cultivated, coagulated
The seed liquor of bacillus is tied, it is spare；Aspergillus niger is taken, is inoculated in aspergillus niger seed liquid culture medium and is cultivated, obtain black song
Mould seed liquor；
Step 5: taking the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, inoculum concentration is fermentation medium volume
2.1%；Then, shaking table culture fermentation, 35 DEG C of temperature, speed 150r/min are placed in；Ferment after 24h, take out, inoculation condenses
The seed liquor of bacillus, inoculum concentration are the 1.8% of fermentation medium；After continuing fermentation 10h, take out, the seed of inoculated aspergillus niger
Liquid, inoculum concentration are the 1.8% of fermentation medium；It is subsequently placed in shaking table culture fermentation, 32 DEG C, speed 200r/min of temperature, fermentation
Time 60h；After resulting zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.
The method that is concentrated of filtering enzymolysis liquid is：Under 0.09 atmospheric pressure, bath temperature reaches 53 DEG C
There is boiling phenomenon, rotating speed 60-80r/min in saccharified liquid.
The seed culture medium of the Lactobacillus casei, 50g containing glucose in every liter of culture medium, 10 g of peptone, beef extract
5 g, 5 g of yeast extract, anhydrous sodium acetate 5 g, MgSO4.7H2O 20mL, Mn SO4.7H2O 20mL, calcium carbonate 20g, pH
6.8 and tween 1mL, surplus are water.
The seed culture culture medium of the bacillus coagulans, 80 g containing glucose in every liter of culture medium, (NH)4SO4 4
G, KH2PO40.3 g, ZnSO4.7H2O 0.44 g, MgSO4.7H20.25 g of O, surplus are water.
The fermentation medium, 50ml containing enzymolysis liquid, peptone 10 g, yeast extract 5g in every liter of culture medium,
MgSO4.7H2O 0. 5 g, (NH)4SO44 g, KH2PO40.4, NaCl 0.1g and CaCO43g, surplus is water.
The method of the Lactic Acid from Fermentation Broth assay is：Analysis Lactic Acid from Fermentation Broth is carried out using high performance liquid chromatography
Content, chromatographic column are the C18 anti-phase (4.6mm × 250mm, 5 μm) of Di Ma companies；Mobile phase is the phosphoric acid of 0.01mol/L
Two ammonium salt solution of hydrogen, with phosphoric acid tune pH values to 2.6, after 0.45 μm of aperture synthetic cellulose esters membrane filtration, ultrasound degassing 1h；Stream
Fast 0.8mL/min, Detection wavelength 210nm, 20 μ L of sample size, column temperature is room temperature, is up to 205g/L, purity in zymotic fluid after testing
Reach 99%.
Raw material of the present invention can be commercially available by market, and bacillus coagulans, Lactobacillus casei, aspergillus niger are
Common strain, experiment proves that common strain can be used, can be commercially available by market；Lactobacillus casei described in embodiment
For Lactobacillus casei CICC6028；Lactobacillus casei CICC6028（Lactobacillus caseiCICC6028）, in being purchased from
The micro- non-hibernating eggs collection of state's industry（CICC）.
Claims (6)
- A kind of 1. method using bagasse production lactic acid, it is characterised in that：Comprise the following steps：Step 1: the pretreatment of bagasse：Bagasse powder is broken to 100-200 mesh, it is spare；With the NaOH that mass fraction is 0.5% Solution carries out the bagasse after crushing sprinkling rubbing, time 12h, bagasse after being pre-processed；Operating method is：Sprinkling Rubbing 5min, is put into 45 DEG C of environment after uniformly；NaOH solution is sprayed every 4h and is rubbed once, rubs 5min every time；Step 2: it is 4.8 to add distilled water in bagasse after the pre-treatment to adjust pH value, zytase and cellulose are then added Enzyme, is put on 50 DEG C of shaking tables and carries out enzymatic saccharification, saccharificatinn period 8h-10h；The addition of the zytase and cellulase The 0.05% and 0.08% of bagasse quality after respectively pre-processing；Step 3: being filtered after saccharification, filtering enzymolysis liquid is obtained；Obtained filtering enzymolysis liquid is concentrated, is obtained Concentrate enzymolysis liquid；Cultivated Step 4: taking Lactobacillus casei to be inoculated in Lactobacillus casei seed liquid culture medium, obtain Lactobacillus casei Seed liquor, it is spare；Take bacillus coagulans to be inoculated in bacillus coagulans seed liquid culture medium to be cultivated, obtain condensing bud The seed liquor of spore bacillus, it is spare；Aspergillus niger is taken, is inoculated in aspergillus niger seed liquid culture medium and is cultivated, obtain aspergillus niger Seed liquor；Step 5: take the seed liquor of Lactobacillus casei to be inoculated in fermentation medium, institute containing step 3 in every liter of fermentation medium Concentration enzymolysis liquid 50ml is stated, Lactobacillus casei seed liquor inoculum concentration is the 2-2.3% of fermentation medium volume；Then, it is placed in shaking table Cultivation and fermentation, 35 DEG C of temperature, speed 150r/min；Ferment after 24h, take out, be inoculated with the seed liquor of bacillus coagulans, inoculum concentration For the 1.5-2% of fermentation medium；After continuing fermentation 10h, take out, the seed liquor of inoculated aspergillus niger, inoculum concentration is fermentation medium 1.5-2%；It is subsequently placed in shaking table culture fermentation, 32 DEG C, speed 200r/min, fermentation time 55h-65h of temperature；Resulting hair After zymotic fluid carries out lactic acid content measure, isolated and purified, that is, obtain lactic acid.
- 2. as claimed in claim 1 using the method for bagasse production lactic acid, it is characterised in that：The filtering enzymolysis liquid carries out The method of concentration is：Under 0.09 atmospheric pressure, bath temperature reaches 53 DEG C of saccharified liquids and boiling phenomenon, rotating speed 60- occurs 80r/min。
- 3. as claimed in claim 1 using the method for bagasse production lactic acid, it is characterised in that：The kind of the Lactobacillus casei Sub- culture medium, 50g containing glucose, peptone 10g, beef extract 5g, yeast extract 5g, anhydrous sodium acetate 5g in every liter of culture medium, MgSO4•7H2O 20mL, MnSO4•7H2O 20mL, calcium carbonate 20g, pH6.8 and tween 1mL, surplus are water.
- 4. as claimed in claim 1 using the method for bagasse production lactic acid, it is characterised in that：The bacillus coagulans Seed culture medium, 80g containing glucose, (NH in every liter of culture medium4)2SO44g, KH2PO40.3g, ZnSO4•7H2O 0.44g, MgSO4•7H2O 0.25g, surplus are water.
- 5. as claimed in claim 1 using the method for bagasse production lactic acid, it is characterised in that：The fermentation medium, often Rise 50ml containing enzymolysis liquid, peptone 10g, yeast extract 5g, MgSO in culture medium4•7H2O 0.5g, (NH4)2SO4 4g, KH2PO4 0.4g, NaCl 0.1g and CaCO3 3g, surplus are water.
- 6. as claimed in claim 1 using the method for bagasse production lactic acid, it is characterised in that：The Lactic Acid from Fermentation Broth contains Measuring method for measuring is：Analysis Lactic Acid from Fermentation Broth content is carried out using high performance liquid chromatography, chromatographic column is the C18 of Di Ma companies It is anti-phase, 4.6mm × 250mm, 5 μm；Mobile phase is the ammonium dibasic phosphate solution of 0.01mol/L, with phosphoric acid tune pH value to 2.6, warp After 0.45 μm of aperture synthetic cellulose esters membrane filtration, ultrasound degassing 1h；Flow velocity 0.8mL/min, Detection wavelength 210nm, sample size 20 μ L, column temperature are room temperature.
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| CN105567567A (en) * | 2016-02-03 | 2016-05-11 | 程雪娇 | Bagasse culture medium and preparing method thereof |
| CN109439698A (en) * | 2018-12-27 | 2019-03-08 | 河南永乐生物工程有限公司 | A kind of method of bacillus coagulans production Pfansteihl |
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| WO2013185344A1 (en) * | 2012-06-15 | 2013-12-19 | Jiangsu University | Production of lactic acid from fermentations using mixed bacterial cultures |
| CN103627739A (en) * | 2013-12-12 | 2014-03-12 | 广西大学 | Method for producing L-lactic acid by fermenting bagasse cellulose |
| CN104694587B (en) * | 2015-02-11 | 2017-12-26 | 北京林业大学 | A kind of method that lactic acid is produced by bagasse |
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