CN109517088A - A method of curdlan is prepared by raw material of sugar-cane juice - Google Patents
A method of curdlan is prepared by raw material of sugar-cane juice Download PDFInfo
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- CN109517088A CN109517088A CN201811325646.1A CN201811325646A CN109517088A CN 109517088 A CN109517088 A CN 109517088A CN 201811325646 A CN201811325646 A CN 201811325646A CN 109517088 A CN109517088 A CN 109517088A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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Abstract
The present invention relates to a kind of methods for preparing curdlan as raw material using sugar-cane juice, adjustment, inoculation fermentation, extraction curdlan of preparation, sugar-cane juice ingredient including sugar-cane juice etc..The present invention is curdlan fermentation primary raw material using from a wealth of sources, cheap sugar-cane juice, reduces production cost, preparation method is simple and easy to do, has a good application prospect.
Description
Technical field
The invention belongs to curdlan field, in particular to a kind of method that curdlan is prepared as raw material using sugar-cane juice.
Background technique
It using glucose is that monomer passes through β -1 that curdlan, which is a kind of, water-insoluble polysaccharide made of 3 glucosides key connections,
Mainly generated by the bacterial fermentation that Agrobacterium or Bacillus foecalis alkaligenes belong to.The polysaccharide has the characteristic of heating solidification plastic, again
Referred to as Curdlan.Cool down again after the aqueous dispersions of curdlan are heated to 55~65 DEG C, forms the minuent of thermal reversibility
Colloid.When the aqueous dispersions of curdlan are heated to 80 DEG C or more, then the height colloid of heat irreversible is formed.Curdlan
It can be widely used for the fields such as food, medicine, chemistry.Because curdlan has safety, at colloidality, U.S. FDA approval in 1996
Curdlan is as a kind of food additives.Curdlan generates biologically active β -1,3 oligomerization after hydrolysis process
Sugar is the desirable feedstock for producing such oligosaccharides.Curdlan is as large biological molecule and a kind of ideal natural polymer material
Material, can be used for the fields such as organizational project.There are antiviral equal immunological characteristics through chemically modified curdlan.
The industrialized production of curdlan is influenced by many factors, and wherein the cost of fermentation medium is wherein important
Factor.The culture medium carbon source of the curdlan of fermentation method production at present is mainly dextrose and saccharose etc..Sucrose be using sugarcane and
Beet is prepared by complicated technology processing.The sugarcane of harvest carries out grinding extraction sugar-cane juice after clearing up, and then adds stone
Ash simultaneously heats to clarify Normal juice, prepares sucrose finally by evaporation and concentration and crystallization etc..Glucose is passed through with starch materials
Hydrolysising purification is prepared.In order to reduce the production cost of curdlan, promote the production and application of curdlan, people attempt
It is produced with starch raw material hydrolysate, plant hydrolyzed liquid.
Sugarcane is a kind of plant that photosynthesis effect is relatively high, and is non-grain crop, and yield is high, and sugared content is high.It is sweet
Sugarcane juice is the juice that sugarcane genotypes generate after squeezing, and for sugar content generally 15~20%, wherein most sugar is sucrose.In addition
It is a kind of good culture substrate containing amino acid, vitamin and several mineral materials.It is culture medium fermentation preparation using sugar-cane juice
Curdlan does not need isolating and purifying for sugar, can reduce production cost, promote the industrialized production of curdlan, has very big
Prospect.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of methods for preparing curdlan as raw material using sugar-cane juice, should
Method solves the prior art and needs additional addition carbon source such as sucrose, glucose etc., causes the problem that fermentation costs are high.
The present invention provides a kind of methods for preparing curdlan as raw material using sugar-cane juice, comprising:
(1) sugar-cane juice and complex culture medium or water are mixed with volume ratio 1:0.1~2, makes the total sugar content 7 of sugar-cane juice
~10% (w/v), sterilizing, obtains the sugar-cane juice through composition adjustment；
(2) curdlan producing strains seed liquor is inoculated in sugar-cane juice through composition adjustment or without the sweet of step (1)
In sugarcane juice (i.e. initial total sugar content meets 7~10% (w/v)), 28~32 DEG C shaken cultivation 3~5 days；
(3) after fermentation, NaOH solution is added in fermentation liquid, stirring evenly is completely dissolved glue；It is then centrifuged for, institute
Obtaining supernatant and adjusting pH value is 7.0, and filters pressing obtains solid curdlan, finally passes through washing, is dehydrated, is dry to get that can obtain so
Glue.
The preparation step of sugar-cane juice in the step (1) is as follows: sugarcane being cleared up and is removed the peel, then sugarcane is sent into and squeezes
It squeezes the juice in machine, gained juice takes supernatant, as sugar-cane juice after being centrifuged；Wherein, centrifugal speed is 4000~6000 revs/min
Clock, centrifugation time are 5~10min.
The ingredient of complex culture medium in the step (1) are as follows: 0.2%~0.4% (w/v) is inorganic nitrogen-sourced, 0.2%~
0.4% (w/v) organic nitrogen source, 0.1% (w/v) MgSO4·7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3。
It is described inorganic nitrogen-sourced for ammonium chloride, dihydrogen phosphate or urea；The organic nitrogen source is yeast extract or corn pulp.
Sterilising temp in the step (1) is 110~120 DEG C, and sterilization time is 20~30min.
The inoculum concentration of curdlan producing strains in the step (2) is 8% (v/v)；Curdlan producing strains are soil
One or more of Bacillus, rhizobium, alcaligenes, bacillus genus, pseudomonas.
Hunting speed in the step (2) is 150~250 revs/min.
The final concentration of 0.5mol/L of NaOH solution in the step (3).
Centrifugal speed in the step (3) is 8000~10000 revs/min；Centrifugation time is 10~20min.
Filters pressing in the step (3) uses autoclave diaphragm filter press.
Dehydration in the step (3) specifically: 1:1 adds dehydrated alcohol dehydration into curdlan by volume.
Drying in the step (3) uses 60 DEG C of heated-air dryings.
Beneficial effect
The present invention directly can prepare curdlan with sugar-cane juice inoculation curdlan production strain, or again with sugar-cane juice
Trophic factors is added to be adjusted, inoculation curdlan production strain prepares curdlan；Compared with prior art, present invention benefit
It is fermenting raw materials production curdlan with the sugar-cane juice that source is wide, preparation is simple, sugar content is high, reduces production cost, and prepare
Method is simple and easy, has a good application prospect.
Detailed description of the invention
Fig. 1 is curdlan infrared spectrogram prepared by embodiment 1；
Fig. 2 is curdlan infrared spectrogram prepared by embodiment 6；
Fig. 3 is the infrared spectrogram of curdlan mark product.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, those skilled in the art
Member can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Range.
Embodiment 1
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 4000 revs/min from
Heart 8min obtains clear sugar-cane juice.
(2) add water by the volume ratio 1:1.5 of sugar-cane juice and water, the sugared content for adjusting sugar-cane juice is 7% (w/v), and 115 DEG C go out
Bacterium 20min；Curdlan producing strains seed liquor is inoculated in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 30 DEG C,
150 revs/min, shaken cultivation 3 days.
(3) fermentation liquid is taken after fermentation, and NaOH solution, which is added, makes its concentration 0.5mol/L, and stirring evenly keeps glue complete
Dissolution, the hydrochloric acid solution tune pH value that 1mol/L is then added in 8000 revs/min of centrifugation 10min, gained supernatant is 7.0,
Then colloid is obtained with autoclave diaphragm filter press filters pressing, after washing 2 times, adds dehydrated alcohol dehydration into colloid by 1:1 volume,
Then in 60 DEG C of dryings.The content of gained curdlan is 20.8g/L.
As shown in Figure 1, the curdlan of sugar-cane juice fermentation preparation is consistent with the infrared spectroscopy height of standard curdlan.
3346cm-1The stretching vibration of the corresponding-OH of neighbouring absorption peak, 2896cm-1Corresponding-the CH of neighbouring absorption peak2Characteristic absorption,
1637cm-1And 1159cm-1Corresponding-the C=O of absorption peak characteristic absorption, 1369cm-1The feature of the corresponding-CH of neighbouring absorption peak
It absorbs, 1034cm-1The characteristic absorption of-C-O, 889cm in the corresponding-C-O-H of neighbouring absorption peak-1Neighbouring absorption peak shows can
In right xanthan molecule D-Glucose with the glycosidic bond links of beta comfiguration.
Embodiment 2
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 6000 revs/min from
Heart 5min obtains clear sugar-cane juice.
(2) add water by sugar-cane juice and water volume ratio 1:1, the sugared content for adjusting sugar-cane juice is 10% (w/v), 115 DEG C of sterilizings
30min；Curdlan producing strains seed liquor is inoculated in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 28 DEG C, 250
Rev/min, shaken cultivation 5 days.
(3) fermentation liquid is taken after fermentation, and NaOH solution, which is added, makes its concentration 0.5mol/L, and stirring evenly keeps glue complete
Then 1mol/L hydrochloric acid solution is added in dissolution in 9000 revs/min of centrifugation 10min, gained supernatant, adjusting pH value is 7.0, so
Solid curdlan is obtained with autoclave diaphragm filter press filters pressing afterwards, after washing 2 times, adds anhydrous second into colloid by 1:1 volume
Dehydration of alcohols, then in 60 DEG C of dryings.The content of gained curdlan is 38.6g/L.
Embodiment 3
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 4000 revs/min from
Heart 10min obtains clear sugar-cane juice.
(2) composite nutrient-fluid: 0.4% (w/v) (NH is prepared4)2Cl, 0.2% (w/v) corn pulp, 0.1% (w/v)
MgSO4·7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3；Sugar-cane juice, 1:1 is mixed by volume with composite nutrient-fluid
It closes, the sugared content for adjusting sugar-cane juice is 8% (w/v), 110 DEG C of sterilizing 30min；
(3) curdlan producing strains seed liquor is inoculated in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 30
DEG C, 150 revs/min shaken cultivation 4 days.
(4) fermentation liquid is taken after fermentation, and NaOH solution is added in fermentation liquid makes its final concentration of 0.5mol/L, and stirring is equal
It is even to be completely dissolved glue, 1mol/L hydrochloric acid solution is then added in 8000 revs/min of centrifugation 15min, gained supernatant, adjusts pH
Value is 7.0, then obtains solid curdlan with autoclave diaphragm filter press filters pressing, after washing 2 times, by 1:1 volume into colloid
Dehydrated alcohol dehydration is added, then in 60 DEG C of dryings.The content of gained curdlan is 40.5g/L.
Embodiment 4
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 6000 revs/min from
Heart 5min obtains clear sugar-cane juice.
(2) composite nutrient-fluid: 0.2% (w/v) urea, 0.2% (w/v) corn pulp, 0.1% (w/v) MgSO is prepared4·
7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3；Sugar-cane juice, 1:0.5 is mixed by volume with composite nutrient-fluid, is adjusted
The sugared content of whole sugar-cane juice is 10% (w/v), 120 DEG C of sterilizing 20min；
(3) curdlan producing strains seed liquor is inoculated in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 32
DEG C, 200 revs/min shaken cultivation 4 days.
(4) fermentation liquid is taken after fermentation, and NaOH solution is added in fermentation liquid makes its final concentration of 0.5mol/L, and stirring is equal
It is even to be completely dissolved glue, 1mol/L hydrochloric acid solution is then added in 10000 revs/min of centrifugation 15min, gained supernatant, adjusts
PH value is 7.0, then obtains solid curdlan with autoclave diaphragm filter press filters pressing, after washing 2 times, by 1:1 volume to colloid
Middle addition dehydrated alcohol dehydration, then in 60 DEG C of dryings.The content of gained curdlan is 47.2g/L.
Embodiment 5
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 5000 revs/min from
Heart 10min obtains clear sugar-cane juice.
(2) composite nutrient-fluid: 0.2% (w/v) (NH is prepared4)2Cl, 0.4% (w/v) yeast extract, 0.1% (w/v)
MgSO4·7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3；Then sugar-cane juice and composite nutrient-fluid by volume 1:
2 mixing, the sugared content for adjusting sugar-cane juice is 9% (w/v), 115 DEG C of sterilizing 20min.
(3) curdlan producing strains are inoculated with seed liquor in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 30
DEG C, 250 revs/min shaken cultivation 4 days.
(4) fermentation liquid is taken after fermentation, and NaOH solution is added in fermentation liquid makes its final concentration of 0.5mol/L, and stirring is equal
It is even to be completely dissolved glue, 1mol/L hydrochloric acid solution is then added in 9000 revs/min of centrifugation 15min, gained supernatant, adjusts pH
Value is 7.0, then obtains solid curdlan with autoclave diaphragm filter press filters pressing, after washing 2 times, by 1:1 volume into colloid
Dehydrated alcohol dehydration is added, then in 60 DEG C of dryings.The content of gained curdlan is 45.5g/L.
Embodiment 6
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 4000 revs/min from
Heart 10min obtains clear sugar-cane juice.
(2) preparation composite nutrient-fluid: 0.4% (w/v) urea, 0.2% (w/v) yeast powder+0.2% (w/v) corn pulp,
0.1% (w/v) MgSO4·7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3；Then sugar-cane juice and composite nutrient-fluid
1:2 is mixed by volume, and the sugared content for adjusting sugar-cane juice is 8% (w/v), 115 DEG C of sterilizing 25min；
(3) curdlan producing strains seed liquor is inoculated in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 30
DEG C, 200 revs/min vibrate 5 days.
(4) fermentation liquid is taken after fermentation, and NaOH solution is added in fermentation liquid makes its final concentration of 0.5mol/L, and stirring is equal
It is even to be completely dissolved glue, 1mol/L hydrochloric acid solution is then added in 10000 revs/min of centrifugation 10min, gained supernatant, adjusts
PH value is 7.0, then obtains solid curdlan with autoclave diaphragm filter press filters pressing, after washing 2 times, by 1:1 volume to colloid
Middle addition dehydrated alcohol dehydration, then in 60 DEG C of dryings.The content of gained curdlan is 44.1g/L.
As shown in Figure 2, the curdlan of sugar-cane juice fermentation preparation adjusted and the infrared spectroscopy of standard curdlan are high
Degree is consistent.3355cm-1The stretching vibration of the corresponding-OH of neighbouring absorption peak, 2896cm-1Corresponding-the CH of neighbouring absorption peak2Spy
Sign absorbs, 1646cm-1And 1159cm-1Corresponding-the C=O of absorption peak characteristic absorption, 1368cm-1Neighbouring absorption peak correspondence-
The characteristic absorption of CH, 1035cm-1The characteristic absorption of-C-O, 890cm in the corresponding-C-O-H of neighbouring absorption peak-1Neighbouring suction
It receives peak and shows that D-Glucose is in curdlan molecule with the glycosidic bond links of beta comfiguration.
Embodiment 7
(1) take sugarcane cleaning peeling, then sugarcane be sent into squeezer in squeeze, gained juice 5000 revs/min from
Heart 8min obtains clear sugar-cane juice.
(2) composite nutrient-fluid: 0.4% (w/v) (NH is prepared4)2HPO4, 0.4% (w/v) corn pulp, 0.1% (w/v)
MgSO4·7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3；Then sugar-cane juice and composite nutrient-fluid by volume 1:
1 mixing, the sugared content for adjusting sugar-cane juice is 10% (w/v), 120 DEG C of sterilizing 20min.
(3) curdlan producing strains seed liquor is inoculated in the sugar-cane juice after sterilizing by the inoculum concentration of 8% (v/v), 32
DEG C, 250 revs/min shaken cultivation 5 days.
(4) fermentation liquid is taken after fermentation, and NaOH solution is added in fermentation liquid makes its final concentration of 0.5mol/L, and stirring is equal
It is even to be completely dissolved glue, 1mol/L hydrochloric acid solution is then added in 10000 revs/min of centrifugation 20min, gained supernatant, adjusts
PH value is 7.0, then obtains solid curdlan with autoclave diaphragm filter press filters pressing, after washing 2 times, by 1:1 volume to colloid
Middle addition dehydrated alcohol dehydration, then in 60 DEG C of dryings.The content of gained curdlan is 49.7g/L.
Claims (10)
1. a kind of method for preparing curdlan as raw material using sugar-cane juice, comprising:
(1) sugar-cane juice and complex culture medium or water are mixed with volume ratio 1:0.1~2, make the total sugar content 7 of sugar-cane juice~
10% (w/v), sterilizing, obtains the sugar-cane juice through composition adjustment；
(2) curdlan producing strains seed liquor is inoculated in sugar-cane juice through composition adjustment or without the sugar-cane juice of step (1)
In, 28~32 DEG C shaken cultivation 3~5 days；
(3) after fermentation, NaOH solution is added in fermentation liquid, stirring evenly is completely dissolved glue；It is then centrifuged for, on gained
It is 7.0 that clear liquid, which adjusts pH value, and filters pressing obtains solid curdlan, finally passes through washing, is dehydrated, is dry to get curdlan.
2. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
The preparation step of sugar-cane juice in step (1) is as follows: sugarcane cleared up and is removed the peel, then sugarcane is sent into squeezer and is squeezed the juice, institute
It obtains juice and takes supernatant, as sugar-cane juice after being centrifuged；Wherein, centrifugal speed is 4000~6000 revs/min, and centrifugation time is
5~10min.
3. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
The ingredient of complex culture medium in step (1) are as follows: 0.2%~0.4% (w/v) is inorganic nitrogen-sourced, and 0.2%~0.4% (w/v) is organic
Nitrogen source, 0.1% (w/v) MgSO4·7H2O, 0.3% (w/v) KH2PO4, 0.3% (w/v) CaCO3。
4. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 3, it is characterised in that: described
Inorganic nitrogen-sourced is ammonium chloride, dihydrogen phosphate or urea；The organic nitrogen source is yeast extract or corn pulp.
5. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
Sterilising temp in step (1) is 110~120 DEG C, and sterilization time is 20~30min.
6. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
The inoculum concentration of curdlan producing strains in step (2) is 8% (v/v)；Curdlan producing strains are Agrobacterium, rhizobium
One or more of category, alcaligenes, bacillus genus, pseudomonas.
7. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
The final concentration of 0.5mol/L of NaOH solution in step (3).
8. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
Centrifugal speed in step (3) is 8000~10000 revs/min；Centrifugation time is 10~20min.
9. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: described
Filters pressing in step (3) uses autoclave diaphragm filter press.
10. a kind of method for preparing curdlan as raw material using sugar-cane juice according to claim 1, it is characterised in that: institute
State the dehydration in step (3) specifically: 1:1 adds dehydrated alcohol dehydration into curdlan by volume.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1583801A (en) * | 2004-06-11 | 2005-02-23 | 江南大学 | Extracting process for microbiological polysaccharide-hot gel |
| CN103695481A (en) * | 2013-12-26 | 2014-04-02 | 广西科学院 | Method for producing succinic acid through fermentation of sugarcane juice |
| CN104017831A (en) * | 2014-06-10 | 2014-09-03 | 河北工业大学 | Method for producing mannitol by taking sugarcane juice as raw material through leuconostoc mesenteroides fermentation |
| CN106701885A (en) * | 2017-02-14 | 2017-05-24 | 华东师范大学 | Fermenting production method of curdlan |
-
2018
- 2018-11-08 CN CN201811325646.1A patent/CN109517088A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1583801A (en) * | 2004-06-11 | 2005-02-23 | 江南大学 | Extracting process for microbiological polysaccharide-hot gel |
| CN103695481A (en) * | 2013-12-26 | 2014-04-02 | 广西科学院 | Method for producing succinic acid through fermentation of sugarcane juice |
| CN104017831A (en) * | 2014-06-10 | 2014-09-03 | 河北工业大学 | Method for producing mannitol by taking sugarcane juice as raw material through leuconostoc mesenteroides fermentation |
| CN106701885A (en) * | 2017-02-14 | 2017-05-24 | 华东师范大学 | Fermenting production method of curdlan |
Non-Patent Citations (2)
| Title |
|---|
| TRIANTARTI DAN HENDRO SANTOSO M: "Effect of Substituting Pure Sucrose by Sugarcane Juice as Carbon Source on the Fermentation of Dextran Production", 《JURNAL ILMU DASAR》 * |
| 董学前: "微生物发酵生产可得然胶的研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
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