CN103626885A - Clean production method of Pulullan - Google Patents
Clean production method of Pulullan Download PDFInfo
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- CN103626885A CN103626885A CN201310564266.4A CN201310564266A CN103626885A CN 103626885 A CN103626885 A CN 103626885A CN 201310564266 A CN201310564266 A CN 201310564266A CN 103626885 A CN103626885 A CN 103626885A
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- pulullan
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- pulullan polysaccharide
- peptone
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Abstract
The invention relates to a clean production method of Pulullan, belonging to the technical field of bioengineering. The method comprises the following steps: fermenting Pulullan-producing microbes in a ventilation environment to obtain a fermentation liquid; heating for inactivation, filtering to remove the thalli, precipitating with an organic solvent, carrying out solid-liquid separation by pressure filtration, and carrying out vacuum drying and pulverization on the solid to obtain the Pulullan product. The clear solution obtained by pressure filtration is rectified to recycle the organic solvent for cyclic use in the next round of precipitation operation. After composition analysis is performed on the residual water after rectification, starch hydrolyzate, peptone, yeast extract, inorganic salt and the like are added to enter the cyclic fermentation medium, and the ventilation fermentation is performed to product the Pulullan fermented liquor, thereby establishing a closed-cycle clean production method. By adjusting the technique, the method implements cyclic utilization of the residual water after rectification under the condition of not increasing equipment and site investment, saves the water consumption, lowers the production cost, reduces the wastewater treatment amount, obviously lowers the treatment amount and discharge amount of phosphorus and nitrogen, and is beneficial to cyclic economy and environmental protection.
Description
Technical field
A clean preparation method for pulullan polysaccharide, belongs to technical field of bioengineering.
Background technology
Pulullan polysaccharide (CAS numbering: 9057-02-7) by Propiram grow sturdily the short stalk of bud mould (
aureobasidium pullulans) microbial polysaccharide that produces through aerobic fermentation, this polysaccharide be the trisaccharide maltose repeating unit that connected by α-Isosorbide-5-Nitrae glycosidic link through α-1, the straight chain shape polysaccharide that 6 glycosidic links are polymerized, molecular weight 20,000～2,000,000 dalton, the polymerization degree 100～5000.Pulullan polysaccharide is high resilience structurally, solubleness is larger, there is film-forming properties, gas barrier property, plasticity-, viscosity all stronger, and there is the good characteristics such as soluble in water, nontoxic, colorless and odorless, be widely used in the fields such as medicine, food, light industry, chemical industry and oil.Within 2006, health ministry approval pulullan polysaccharide is foodstuff additive, can in candy, chocolate-coated, diaphragm, compound seasoner and juice drinks, be used as fruit glaze agent and thickening material, and usage quantity standard can be according to GB2760-2011.At pharmaceutical industries pulullan polysaccharide, can be used for manufacturing capsule, lozenge, amorphism medicine, hemostatic agent, blood plasma extender, radioscopy contrast medium; Toxoid and vaccine protective material; Wound suture line; Artificial organ and anticoagulant property medical material etc.Propiram capsule is compared with other capsule, has very significantly advantage: 1. the oxygen permeability of Propiram capsule is approximately 1/8 of gelatine capsule, can effectively protect content to avoid oxidation, extends shelf time; 2. transparency is suitable with animality capsule, and filling content is clearly visible; 3. not containing animal protein and fat, be difficult for growing microorganism, steady quality; 4. not containing animality composition, without the secret worry of the common transmissible diseases of people and animals such as mad cattle disease, foot and mouth disease; 5. natural plant raw material is made, the restriction of irreligion and vegetarian diet aspect.At cosmetic industry, can be used for manufacturing face powder, shampoo, astringent, facial mask, Derma-Guard, hairspray etc.
The production process of pulullan polysaccharide is mainly made by unit operations such as ventilating fermentation, heat inactivation, filtration sterilization, organic solvent deposit, vacuum-drying, solvent recuperation and wastewater treatments.Owing to having used the organic solvents such as a large amount of ethanol or Virahol, for meeting cost economy and environmental requirement, must reclaim solvent in process of production.As reclaim 1 ton of ethanol approximately produce 0.6 ~ 0.9 ton more than heat up in a steamer water, reclaim 1 ton of Virahol and approximately produce more than 1.2 ~ 1.5 tons and heat up in a steamer water.More than heat up in a steamer phosphorus (total phosphorus >10 mg/L) and the nitrogen (total nitrogen is 200 ~ 500mg/L) that is rich in high density in water, COD is up to 7000 ~ 20000 mg/L, BOD is up to 5000 ~ 15000 mg/L, bring heavier processing load and Financial cost to the wastewater treatment of follow-up operation, moreover be rich in the phosphoric of high density, be also unfavorable for the efficient processing of organic waste water.If heat up in a steamer water more than Efficient Cycle utilization in process of production, will improve on the one hand the utilization ratio of raw material, reduce production costs and energy consumption, produce good economic benefit; Also can reduce on the other hand processing cost and the quantity discharged of factory effluent, promote the green manufacturing of pulullan polysaccharide, produce good environmental benefit and social benefit.
Summary of the invention
The clean preparation method that the object of this invention is to provide a kind of microbial polysaccharide pulullan polysaccharide, to reduce sewage load and the quantity discharged in pulullan polysaccharide production process, save raw materials for production and energy consumption simultaneously, reduce the production cost of microbial polysaccharide, make its market competitiveness stronger.
Technical scheme of the present invention: a kind of clean preparation method of pulullan polysaccharide, utilize and produce pulullan polysaccharide microorganism through the fermented liquid of ventilating fermentation gained, through following processing step, process:
(1) heat inactivation: make microorganism and enzyme deactivation through 60 ~ 100 ℃ of heat treated 10-20 minute;
(2) filtration sterilization body: adopt Plate Filtration to process pulullan polysaccharide fermented liquid to remove the microbial cells including, obtain the filtrate A of clarification;
(3) with organic solvent deposit: add with the ethanol of 0.5 ~ 2.5 times of volume of filtrate A or Virahol or acetone and carry out precipitation process, precipitation operation temperature is 10 ~ 40 ℃, and sedimentation time is 4 ~ 10 hours;
(4) centrifugal or press filtration is separated solid-liquid two: through centrifugation or squeezing, process, obtain solid substance D and supernatant liquor B; Solid substance D uses a small amount of above-mentioned corresponding ethanol or Virahol or washing with acetone 1～2 time again, ethanol used or Virahol or amounts of acetone are 0.3～1 times of solid substance D volume, washing operation temperature is 10 ~ 40 ℃, then centrifugal or squeezing removes organic solvent and obtains supernatant C and solid substance E, and solid substance E obtains pulullan polysaccharide product after temperature is no more than 85 ℃ of vacuum-dryings and pulverizes;
(5) rectifying is reclaimed: supernatant liquor B is incorporated to distillation system and reclaims together with supernatant C, and the ethanol obtaining through distillation operation recovery or Virahol or acetone recycle, and the remaining distillate that distillation operation produces is included in circulating fermentation substratum;
(6) remaining distillate reuse: supplement appropriate starch hydrolyzate, peptone, yeast extract, a small amount of phosphoric acid salt in the remaining distillate after step (5) is processed, supply process water to the volume requiring, adjust pH value to neutral, after intermittence or continuous sterilization, as circulating fermentation substratum, enter fermentor tank and produce pulullan polysaccharide for circulating fermentation, it is 3 ~ 5 times that remaining distillate recycles number of times, is built into the clean preparation method of a closed cycle.
Total reducing sugar to remaining distillate, total nitrogen, total phosphorus, COD, BOD, the index such as colourity and specific conductivity is carried out after quantitative analysis, described in step (6), in remaining distillate, supplement appropriate starch hydrolyzates, peptone, yeast extract, phosphoric acid salt and water, the main component that makes circulating fermentation substratum and initial fermention medium component content and index are quite or equate, the amount of supplementary starch hydrolyzates be in initial fermention medium starch hydrolyzates amount 50% ~ 90%, peptone and amount of yeast extract be in initial fermention medium peptone and amount of yeast extract 65% ~ 90%, amount of phosphate be in initial fermention medium amount of phosphate 5% ~ 40%.
The sterilization process condition of circulating fermentation substratum is intermittently sterilizing in batches in 115 ~ 130 ℃, 20 ~ 40 minutes, or 120 ~ 140 ℃, 200 ~ 400 seconds continuous sterilizations; The follow-up install pipeline of sterilizing has filtration unit.
Beneficial effect of the present invention: pulullan polysaccharide product of the present invention is food grade.Prior art adopts filtration sterilization, solvent flocculation sediment, solvent wash and vacuum drying method.Its preparation process is used a large amount of organic solvents, produces more than a large amount of and heat up in a steamer water in reclaiming the process of solvent, has both wasted a large amount of heats and water, brings pressure again to wastewater treatment, makes the production cost of pulullan polysaccharide high.The present invention is grouped into the nutritional needs with fermention medium by heating up in a steamer the one-tenth of water more than relatively, by remaining, heat up in a steamer in the formula that water is integrated into fermention medium, built the utilization process of pulullan polysaccharide extraction waste water, cycle index is 3 ~ 5 times, more than heat up in a steamer water quantity discharged and be reduced to 60% ~ 70% of former technique, the water consumption of fermenting process substratum preparation is reduced to 30% ~ 40% of former technique, more than reclaiming, heat up in a steamer the heat of water simultaneously, reduce the usage quantity of the culture medium raw materials such as main raw material starch hydrolyzates, peptone, yeast extract and phosphoric acid salt.The present invention is different from the production technique of original pulullan polysaccharide, can reduce water consumption, steam consumption, culture medium raw material usage quantity and cost for wastewater treatment in production process, can reduce production costs 8% ~ 15%, for newly-built factory, can reduce construction investment and the running cost of wastewater treatment.Enforcement of the present invention will produce obvious environmental benefit and social benefit.The food grade pulullan polysaccharide of gained can be used as the additive of the food such as candy, chocolate-coated, diaphragm, compound seasoner and garden spgarden stuff, and to improve the characteristic of these food, in foodstuffs industry, tool has been widely used.
Accompanying drawing explanation
The technological process of production schematic diagram of Fig. 1 pulullan polysaccharide.
Embodiment
Embodiment 1: the preparation of pulullan polysaccharide fermented liquid
Produce pulullan polysaccharide bacterial strain and be Propiram grow sturdily the short stalk of bud mould (
aureobasidium pullulans) preservation of key lab of the industrial biotechnology the Ministry of Education of B-1You Southern Yangtze University, Wuxi Light Industry Univ.'s journal, 2003,22 (1): 16-20 is open.
Seed culture medium (kg/m
3): sucrose 20; Peptone 2.0; Extractum carnis 3.0; Yeast extract 2.0, prepares with tap water; PH 7.0.
Initial fermention medium (kg/m
3): sucrose 50; Yeast extract 2.0; (NH
4)
2sO
41.5; K
2hPO
42.0; KH
2pO
41.5; MgSO
47H
20 1.0; With tap water, prepare; PH 7.0.
Fermentation condition: fermenting process is implemented at the fermentor tank of 5 cubic metres, inoculum size 5% (v/v), mixing speed 300 r/min, ventilation is 1.0 vvm, tank pressure 0.05 MPa, 25 ℃ of leavening temperatures.When fermentation starts, pH declines naturally, and the NaOH solution with 2.0 mol/L after 18h is constant in 5.0 by pH value, and fermentation culture 7 is tank all over the world, measures pulullan polysaccharide output and quality-controlling parameters.
Embodiment 2: the product preparation of pulullan polysaccharide
Measure 1000 liters of pulullan polysaccharide fermented liquids, according to fermentation broth viscosity, add 2.0 times of deionized water dilutions, steam jacket is heated to 85 ℃, after maintaining 15 minutes, be premixed into 0.5 ~ 1.5kg diatomite (particle size is 200 ~ 300 orders), adopt flame filter press to filter (filter cloth aperture is 500 ~ 600 orders), working pressure is 0.14-0.24 MPa, removes microbial cells, obtains filtrate A.
In the filtrate A of gained, slowly add 3000 liters of alcohol solvents, after stirring and evenly mixing standing 30 minutes, adopt expeller separated (2000 rpm), obtain about 280kg solid substance D, supernatant liquor B is ethanolic soln, is delivered to waste ethanol basin.Gained solid substance D is delivered in washing tank, add 500 liters of alcohol solvents, after stirring and evenly mixing standing 30 minutes, again adopt expeller separated (2000 rpm), the 170kg solid substance E obtaining is delivered in Vacuumdrier, the supernatant C obtaining is ethanol waste liquid, is delivered to waste ethanol basin and concentrates Distillation recovery to process.The solid substance E of above-mentioned gained vacuum-drying 3 ~ 6 hours under lower than 80 ℃ of conditions, the solution of vacuum condensation gained is delivered to waste ethanol basin, dry after gained solid substance through pulverize and screening after obtain pulullan polysaccharide product.
Embodiment 3: the preparation of the recycle of remaining distillate and circulating fermentation substratum
The waste ethanol of gained in embodiment 2 (supernatant liquor B and supernatant C) is concentrated and is taken in to waste ethanol basin, and the solvent deposition that the ethanol of gained can be back to next round circulation pulullan polysaccharide after rectifying is with separated.The remaining distillate of gained is 1800 liters, the substratum preparation of the workshop section that can be applicable to ferment after composition analysis.
The composition that obtains remaining distillate through chemical analysis is as following table:
Contents of Main Components in distillate more than table 1
| Sample | COD(mg/L) | BOD(mg/L) | Total reducing sugar (g/L) | Total nitrogen (g/L) | Total phosphorus (g/L) | Magnesium ion (g/L) |
| Remaining distillate | 6000 | 5600 | 1.75 | 0.075 | 0.34 | 0.042 |
Circulating fermentation substratum preparation: replace tap water preparation fermention medium (kg/m with the remaining distillate (approximately 70 ℃) of heat
3), starch hydrolyzates (sugar) 42; Yeast extract 2.0; (NH
4)
2sO
41.2; K
2hPO
41.20; KH
2pO
40.80; MgSO
47H
2o 0.7; PH 7.0;
Remaining distillate and tap water proportioning are 4:1.
Press the treatment process described in embodiment 2, obtain more than 1800 liters distillate, during the preparation of circulating fermentation substratum, with tap water, supplement.When recycle number of times is 4 times, substantially do not affect the quality product of pulullan polysaccharide.
Claims (3)
1. a clean preparation method for pulullan polysaccharide, is characterized in that utilizing and produces pulullan polysaccharide microorganism through the fermented liquid of ventilating fermentation gained, through following processing step, processes:
(1) heat inactivation: make microorganism and enzyme deactivation through 60 ~ 100 ℃ of heat treated 10-20 minute;
(2) filtration sterilization body: adopt Plate Filtration to process pulullan polysaccharide fermented liquid to remove the microbial cells including, obtain the filtrate A of clarification;
(3) with organic solvent deposit: add with the ethanol of 0.5 ~ 2.5 times of volume of filtrate A or Virahol or acetone and carry out precipitation process, precipitation operation temperature is 10 ~ 40 ℃, and sedimentation time is 4 ~ 10 hours;
(4) centrifugal or press filtration is separated solid-liquid two: through centrifugation or squeezing, process, obtain solid substance D and supernatant liquor B; Solid substance D uses a small amount of above-mentioned corresponding ethanol or Virahol or washing with acetone 1～2 time again, ethanol used or Virahol or amounts of acetone are 0.3～1 times of solid substance D volume, washing operation temperature is 10 ~ 40 ℃, then centrifugal or squeezing removes organic solvent and obtains supernatant C and solid substance E, and solid substance E obtains pulullan polysaccharide product after temperature is no more than 85 ℃ of vacuum-dryings and pulverizes;
(5) rectifying is reclaimed: supernatant liquor B is incorporated to distillation system and reclaims together with supernatant C, and the ethanol obtaining through distillation operation recovery or Virahol or acetone recycle, and the remaining distillate that distillation operation produces is included in circulating fermentation substratum;
(6) remaining distillate reuse: supplement appropriate starch hydrolyzate, peptone, yeast extract, a small amount of phosphoric acid salt in the remaining distillate after step (5) is processed, supply process water to the volume requiring, adjust pH value to neutral, after intermittence or continuous sterilization, as circulating fermentation substratum, enter fermentor tank and produce pulullan polysaccharide for circulating fermentation, it is 3 ~ 5 times that remaining distillate recycles number of times, is built into the clean preparation method of a closed cycle.
2. clean preparation method according to claim 1, it is characterized in that the total reducing sugar to remaining distillate, total nitrogen, total phosphorus, COD, BOD, colourity and conductivity indices are carried out after quantitative analysis, described in step (6), in remaining distillate, supplement appropriate starch hydrolyzates, peptone, yeast extract, phosphoric acid salt and water, the main component that makes circulating fermentation substratum and initial fermention medium component content and index are quite or equate, the amount of supplementary starch hydrolyzates be in initial fermention medium starch hydrolyzates amount 50% ~ 90%, peptone and amount of yeast extract be in initial fermention medium peptone and amount of yeast extract 65% ~ 90%, amount of phosphate be in initial fermention medium amount of phosphate 5% ~ 40%.
3. clean preparation method according to claim 1, the sterilization process condition that it is characterized in that circulating fermentation substratum is intermittently sterilizing in batches in 115 ~ 130 ℃, 20 ~ 40 minutes, or 120 ~ 140 ℃, 200 ~ 400 seconds continuous sterilizations; The follow-up install pipeline of sterilizing has filtration unit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310564266.4A CN103626885A (en) | 2013-11-14 | 2013-11-14 | Clean production method of Pulullan |
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|---|---|---|---|
| CN201310564266.4A CN103626885A (en) | 2013-11-14 | 2013-11-14 | Clean production method of Pulullan |
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| CN103626885A true CN103626885A (en) | 2014-03-12 |
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ID=50208359
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112707940A (en) * | 2020-12-28 | 2021-04-27 | 中国科学院青岛生物能源与过程研究所 | Sterilization method of glycerol glucoside |
| CN113735991A (en) * | 2021-10-11 | 2021-12-03 | 天津科技大学 | Process for extracting pullulan from high-viscosity fermentation liquor |
| US11319566B2 (en) | 2017-04-14 | 2022-05-03 | Capsugel Belgium Nv | Process for making pullulan |
| US11576870B2 (en) | 2017-04-14 | 2023-02-14 | Capsugel Belgium Nv | Pullulan capsules |
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| CN1654482A (en) * | 2005-02-21 | 2005-08-17 | 汕头市富味制果厂有限公司 | Process for producing pullulan |
| CN101560261A (en) * | 2009-05-11 | 2009-10-21 | 江南大学 | Cleaner production method of gellan gum |
| WO2012081616A1 (en) * | 2010-12-14 | 2012-06-21 | Jsr株式会社 | Method for producing polymer particles, and polymer particles |
| CN103172757A (en) * | 2012-12-31 | 2013-06-26 | 天津北洋百川生物技术有限公司 | Extracting process of pulullan polysaccharide |
-
2013
- 2013-11-14 CN CN201310564266.4A patent/CN103626885A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651467A (en) * | 2005-01-11 | 2005-08-10 | 苏理 | Solvent extraction production technology of pullulanose |
| CN1654482A (en) * | 2005-02-21 | 2005-08-17 | 汕头市富味制果厂有限公司 | Process for producing pullulan |
| CN101560261A (en) * | 2009-05-11 | 2009-10-21 | 江南大学 | Cleaner production method of gellan gum |
| WO2012081616A1 (en) * | 2010-12-14 | 2012-06-21 | Jsr株式会社 | Method for producing polymer particles, and polymer particles |
| CN103172757A (en) * | 2012-12-31 | 2013-06-26 | 天津北洋百川生物技术有限公司 | Extracting process of pulullan polysaccharide |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11319566B2 (en) | 2017-04-14 | 2022-05-03 | Capsugel Belgium Nv | Process for making pullulan |
| US11576870B2 (en) | 2017-04-14 | 2023-02-14 | Capsugel Belgium Nv | Pullulan capsules |
| US11878079B2 (en) | 2017-04-14 | 2024-01-23 | Capsugel Belgium Nv | Pullulan capsules |
| CN112707940A (en) * | 2020-12-28 | 2021-04-27 | 中国科学院青岛生物能源与过程研究所 | Sterilization method of glycerol glucoside |
| CN112707940B (en) * | 2020-12-28 | 2022-07-01 | 中国科学院青岛生物能源与过程研究所 | Sterilization method of glycerol glucoside |
| CN113735991A (en) * | 2021-10-11 | 2021-12-03 | 天津科技大学 | Process for extracting pullulan from high-viscosity fermentation liquor |
| CN113735991B (en) * | 2021-10-11 | 2022-06-03 | 天津科技大学 | Process for extracting pullulan from high-viscosity fermentation liquor |
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Application publication date: 20140312 |