CN101736042B - Method for producing L-lactic acid - Google Patents
Method for producing L-lactic acid Download PDFInfo
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Abstract
The invention relates to a method for producing L-lactic acid. In the method, crop straw and Rhizopus oryzae serve as raw materials. The method particularly comprises the following operation steps: A. preparing straw saccharified liquid; B. adding auxiliary material to inoculate Rhizopus oryzae for fermentation; C. repeatedly fermenting 25 batches; D. acidizing fermented clear liquid; E. ultrafiltration; F. nanofilteration; G. reverse osmosis; H. ion exchange; I. concentrating in vacuum to obtain the L-lactic acid product with the mass percentage concentration of 85%. The invention takes the straw hydrolysis saccharified liquid as the main raw material to prepare L-lactic acid, which obviously lowers the production cost of L-lactic acid; Rhizopus oryzae is adopted for once inoculation proliferation, thallus can be reused for several times to produce L-lactic acid, thus shortening L-lactic acid fermentation period, and obviously improving the fermentation strength of L-lactic acid; an advanced membrane separation and ion exchange integration technology is adopted to improve the lactic acid product quality; and the invention has simple process and strong continuity.
Description
Technical field
The present invention relates to a kind of method of the L-of production lactic acid, particularly relate to a kind of Rhizopus oryzae (Rhizopus oryzae) AS3.819 and utilize the agricultural crop straw hydrolysis sugar liquid, L-lactic acid and membrane sepn and the integrated method of separating L-lactic acid of ion-exchange are produced in thalline recycling fermentation.
Background technology
Lactic acid has been widely used in food, medicine, medical science and chemical engineering industry as a kind of important organic acid.In recent years, all over the world in, find that with L-lactic acid be the plastics film that the polymkeric substance-poly (l-lactic acid) of the L-lactic acid that forms of monomer polymerization is made, can 100% be biological degradation for the research that solves " white pollution " that plastics brought.Human body can only metabolism L-lactic acid, and the World Health Organization advocates at food, field of medicaments, uses L-lactic acid, does not use DL-lactic acid.The demand of L-lactic acid constantly enlarges.
Lactic acid can obtain by the fermentation of chemosynthesis or carbohydrate, can the narrow spectrum L-of obtaining lactic acid or D-lactic acid but be based on fermentation method, so fermentation method is widely adopted the production of L-lactic acid.The Rhizopus oryzae fermenting lactic acid has the optical purity height, thalline easily with characteristics such as fermented liquid separates, utilize Rhizopus oryzae fermentation to become the trend of scale operation L-lactic acid.
In order to reduce the preparation cost of L-lactic acid, improve the lactic acid-producing quality, lactic acid production process has been carried out big quantity research both at home and abroad, mainly concentrate on: the development and use of low price raw material, improve ferment strength, optimize these three aspects of lactic acid separation-extraction technology.The raw material that is used for the L-lactic acid-producing at present mainly contains starch, molasses, whey water etc.About altogether 1,000,000,000 tons of the lignocellulosic material that annual waste of China or low-quality are utilized, wherein one of agricultural crop straw is with regard to nearly more than 700,000,000 tons, and its major ingredient is fiber, half fiber, xylogen.
L-lactic acid fermentation liquid complicated component except that L-lactic acid, also contains impurity such as bacterial chip, protein, pigment, residual sugar and fermentation byproduct.The traditional separation method of lactic acid has crystallization processes and the direct acidolysis process of calcium lactate before the calcium lactate, and process is simple, is easy to control, be widely adopted, but the lactic acid yield is low, and only about 80%, product purity is low, and many impurity can't effectively be separated in the fermented liquid.Based on this, Chinese scholars has been done a large amount of research to the separation of lactic acid, proposes process separation and purification lactic acid such as reaction, extraction, absorption, common distillation, molecular distillation.Seek an economy, separating and purifying method efficiently, improve the quality and the yield of product, reduce production costs,, become the key that can lactic acid and polymkeric substance thereof large-scale promotion application, have great meaning to adapt to the needs of scale operation.
Membrane separation technique as a novel high score from, concentrate, extraction and purification techniques.Developing over nearly 30 years rapidly, in each industrial circle and scientific research, be widely used.Membrane separation technique does not relate to phase transformation in the process of separate substance, to energy require low, therefore need the process of intake to compare with distillation, crystallization, evaporation etc. very big difference is arranged, the condition of membrane sepn is generally relatively gentleer, for separation, the classification of heat-sensitive substance, complex material, concentrate and enrichment has extremely important meaning.Domestic existing report adopts the extractive technique of membrane separation technique to lactic acid fermentation liquid.The patent " a kind of process for cleanly preparing of lactic acid " (application number 200810195297.6) of Nanjing University of Technology's application in 2008 has been reported a kind of lactic acid extraction technology of utilizing membrane filtration and bipolar membrane electrodialysis; Jiangsu Daosen Biochemical Co., Ltd. had applied for patent " a kind of novel process of separation and purification L-lactic acid " (application number: 200810020397.5) reported a kind of novel process of utilizing continuous from friendship, membrane sepn, molecular distillation technique separation and purification L-lactic acid in 2008, L-lactic acid is adsorbed onto on the anionite-exchange resin earlier, again through washing assorted, wash-out operation, produce a large amount of waste water, be unfavorable for operate continuously.In present L-lactic acid production process, adopt glucose, starch etc. as fermenting carbon source, the raw materials cost height more; Adopt batch fermentation, each batch fermentation period is long, and ferment strength is low; Follow-up separation process complexity, continuity are not strong.
Summary of the invention
In order to reduce the lactic acid-producing material cost, improve ferment strength, to overcome in the existing lactic acid separation method continuity not strong, deficiencies such as process complexity, the invention provides with agricultural crop straw and Rhizopus oryzae is the production method of a kind of L-lactic acid of raw material.
The technical solution that realizes above-mentioned purpose is as follows:
A kind of method of the L-of production lactic acid comprises following operation steps:
(1) the agricultural crop straw maize straw is pulverized the back and cross 40 mesh sieves, employing sig water, diluted acid, cellulase are handled successively, make to contain the saccharification liquid 10L that saccharic amount per-cent is 8-12%;
(2) be that 8-12% saccharification liquid adds in the fermentor tank with the mass percent that makes, add lime carbonate 400-600g, ammonium nitrate 30-35g, potassium primary phosphate 3-5g, SODIUM PHOSPHATE, MONOBASIC 2-3g, zinc sulfate 3-4g, sal epsom 3-4g and ferrous sulfate 0.8-1.0g simultaneously, sterilization is cooled to 32 ℃～34 ℃, and inoculating spores concentration is 10
11The Rhizopus oryzae of individual/L (Rhizopusoryzae) AS3.819 spore suspension 500mL carries out aeration-agitation fermentation, fermentation time 60-65h at 32 ℃～34 ℃; Stop to ventilate and stirring, remaining sugar concentration leaves standstill less than 5g/L in jar; Rhizopus oryzae thalline natural subsidence arrives at the bottom of the fermentor tank, adopts malleation to discharge the L-lactic acid fermentation liquid of 5/6 volume;
(3) set by step (2) add material again in fermentor tank, carry out the aeration-agitation fermentation at 32 ℃～34 ℃, repeat 25 batches, each batch fermentation time 20～22h; Each batch fermentation ends stops to ventilate and stirs, recycling at the bottom of the Rhizopus oryzae thalline is stayed jar, and malleation is emitted the L-lactic acid fermentation liquid of 5/6 volume;
(4) the L-lactic acid fermentation liquid that step (2) and (3) are obtained merges, than adding 4.5 ‰ (w/v) calcium hydroxide, 7.5 ‰ (w/v) gacs and 3 ‰ (w/v) sal epsom, stir 45min down by mass/volume, leave standstill at 70 ℃, impurity is removed in centrifuging, obtains the L-lactic acid fermentation clear liquid;
(5) adopt 98% vitriol oil that the L-lactic acid fermentation clear liquid is carried out acidifying, staticly settle, remove calcium sulfate, obtain acidifying fermentation liquid;
(6) be 3000 daltonian rolling ultra-filtration membranes with acidifying fermentation liquid by molecular weight cut-off, working pressure 0.8Mpa, concentration ratio are 10, remove macromole impurity such as deproteinize, obtain ultrafiltration and concentration liquid and ultrafiltration and see through liquid, ultrafiltration and concentration liquid filters after handling through charcoal absorption again;
(7) ultrafiltration is seen through liquid and adopt nanofiltration membrane to filter, molecular weight cut-off 150～300 dalton, working pressure 1.0MPa accounts for 95% of feed volume through liquid is long-pending, obtains nanofiltration concentrated solution and nanofiltration and sees through liquid, and the nanofiltration concentrated solution enters the ultra-filtration membrane ultrafiltration;
(8) nanofiltration is seen through liquid and adopt the reverse osmosis pre-concentration, working pressure 4MPa obtains massfraction and is 18% L-lactic acid pre-concentration liquid;
(9) L-lactic acid pre-concentration liquid is removed inorganic salt through storng-acid cation exchange resin, weak base anion-exchange resin successively, obtain desalination L-lactic acid solution;
(10) desalination L-lactic acid solution enters the vacuum distilling unit and concentrates, and obtains the L-lactic product of mass percentage concentration 85%, and quality reaches the GB2023-2003 standard.
Described agricultural crop straw is maize straw or wheat stalk or rice straw.
The present invention adopts Rhizopus oryzae fermentation agricultural crop straw hydrolysis sugar liquid, and membrane technique and the integrated purification L-lactic acid that separates of ion-exchange are adopted in the thalline recycling, and its useful technique effect is embodied in:
(1) the present invention is that main raw material prepares L-lactic acid with a large amount of agricultural crop straw hydrolysis sugar liquids, significantly reduces the production cost of L-lactic acid; The stalk hydrolyzed solution is five hexose mixed solutions, and Rhizopus oryzae can utilize five hexoses simultaneously, utilizes stalk hydrolysis sugar liquid mass preparation lactic acid, and its industrial prospect is boundless.
(2) the present invention adopts Rhizopus oryzae once to inoculate propagation, thalline repeatedly reuses produces L-lactic acid, shorten the L-lactic fermentation cycle, the batch fermentation that present most of producer adopts, fermentation period is about 72 hours, it is 60-72h that the present invention removes first batch fermentation time, and follow-up repetition batch fermentation time is 20-22 hour, and the ferment strength of L-lactic acid significantly improves.
(3) the present invention adopts advanced membrane sepn, ion-exchange integrated technology, has improved the quality product of lactic acid.Traditional ion-exchange isolation technique is that lactate ion is adsorbed onto on the anion-exchange column by anionresin, takes off by pickling to obtain lactic acid solution again, and the process complexity produces a large amount of waste water.The present invention will remove the lactic acid solution that albumen, nanofiltration takes off after divalent salts and pigment, the reverse osmosis concentration through ultrafiltration and remove remaining positively charged ion through Zeo-karb by cationic exchange earlier, then feed the weak base anion-exchange resin post, exchange absorption sulfate ion, chlorion etc., effluent liquid concentrates and promptly obtains lactic product, process is simple, and continuity is strong.
Description of drawings
Fig. 1 is a kind of technological process block-diagram of producing L-lactic acid.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
Specific embodiments:
Embodiment 1:
Press shown in the accompanying drawing 1, operation steps of the present invention is as follows:
(1) maize straw is pulverized the back and cross 40 mesh sieves, ratio in 5L/Kg adds calcium hydroxide aqueous solution, behind 95 ℃ of following heated and stirred 25min, add in 98% vitriol oil extremely neutral with calcium hydroxide, separate and remove solid composition and calcium sulfate precipitation, add enzyme in the ratio of 1g/L and live and be the liquid cellulase of 15U/g, 60 ℃ of stirring enzymolysis 60min down make that to contain saccharic amount per-cent be 10% saccharification liquid 10L;
(2) be that 8-12% saccharification liquid adds in the fermentor tank with the mass percent that makes, add lime carbonate 500g, ammonium nitrate 33g, potassium primary phosphate 4g, SODIUM PHOSPHATE, MONOBASIC 2.5g, zinc sulfate 3.5g, sal epsom 3.5g and ferrous sulfate 0.9g simultaneously, sterilization is cooled to 32 ℃-34 ℃, and inoculating spores concentration is 10
11The Rhizopus oryzae of individual/L (Rhizopus oryzae) AS3.819 spore suspension 500mL carries out aeration-agitation fermentation, fermentation time 62h at 32 ℃-34 ℃; Stop to ventilate and stirring, remaining sugar concentration leaves standstill less than 5g/L in jar; Rhizopus oryzae thalline natural subsidence arrives at the bottom of the fermentor tank, adopts malleation to discharge the L-lactic acid fermentation liquid of 5/6 volume;
(3) set by step (2) add material again in fermentor tank, carry out the aeration-agitation fermentation at 32 ℃-34 ℃, repeat 25 batches, each batch fermentation time 21h; Each batch fermentation ends stops to ventilate and stirs, recycling at the bottom of the Rhizopus oryzae thalline is stayed jar, and malleation is emitted the L-lactic acid fermentation liquid of 5/6 volume;
(4) the L-lactic acid fermentation liquid that step (2) and (3) are obtained merges, than adding 4.5 ‰ (w/v) calcium hydroxide, 7.5 ‰ (w/v) gacs and 3 ‰ (w/v) sal epsom, stir 45min down by mass/volume, leave standstill at 70 ℃, impurity is removed in centrifuging, obtains the L-lactic acid fermentation clear liquid;
(5) adopt 98% vitriol oil that the L-lactic acid fermentation clear liquid is carried out acidifying, the material temperature is no more than 80 ℃, check with 0.1% methyl violet solution, the acidolysis equivalent is safran when suitable, be green during excess sulfuric acid, be purple when calcium lactate is excessive, add calcium lactate accordingly when sulfuric acid or lactic acid are excessive or sulfuric acid neutralizes.Leave standstill after the neutralization fully and calcium sulfate was fully separated out in 2 hours, remove calcium sulfate, obtain acidifying fermentation liquid;
(6) be 3000 daltonian rolling ultra-filtration membranes with acidifying fermentation liquid by molecular weight cut-off, working pressure 0.8Mpa, concentration ratio are 10, remove macromole impurity such as deproteinize, obtain ultrafiltration and concentration liquid and ultrafiltration and see through liquid, ultrafiltration and concentration liquid filters after handling through charcoal absorption again;
(7) ultrafiltration being seen through liquid adopts the DK type nanofiltration membrane of General Electric (GE) company to filter, molecular weight cut-off 150-300 dalton, working pressure 1.0MPa sees through long-pending 95% of the feed volume that accounts for of liquid, obtain nanofiltration concentrated solution and nanofiltration and see through liquid, the nanofiltration concentrated solution enters the ultra-filtration membrane ultrafiltration;
(8) nanofiltration is seen through liquid and adopt the reverse osmosis pre-concentration, reverse osmosis membrane is the SG type reverse osmosis membrane of General Electric (GE) company, and working pressure 4MPa obtains massfraction and be 18% L-lactic acid pre-concentration liquid;
(9) L-lactic acid pre-concentration liquid is removed inorganic salt through storng-acid cation exchange resin, weak base anion-exchange resin successively, obtain desalination L-lactic acid solution;
(10) desalination L-lactic acid solution enters the vacuum distilling unit and concentrates, and obtains the L-lactic product of mass percentage concentration 85%, and it is 99% that L-lactic acid accounts for total lactic acid ratio, and quality reaches the GB2023-2003 standard.
Embodiment 2:
(1) wheat stalk is pulverized the back and cross 40 mesh sieves, ratio in 5.5L/Kg adds calcium hydroxide aqueous solution, behind 95 ℃ of following heated and stirred 20min, add in 98% vitriol oil extremely neutral with calcium hydroxide, separate solid composition and calcium sulfate precipitation, add enzyme in the ratio of 0.8g/L and live and be the liquid cellulase of 15U/g, 60 ℃ of stirring enzymolysis 45min down make that to contain saccharic amount per-cent be 8% saccharification liquid 10L;
(2) be that 8% saccharification liquid adds in the fermentor tank with the mass percent that makes, add lime carbonate 400g, ammonium nitrate 30g, potassium primary phosphate 3g, SODIUM PHOSPHATE, MONOBASIC 2g, zinc sulfate 3g, sal epsom 3g and ferrous sulfate 0.8g simultaneously, sterilization is cooled to 32 ℃-34 ℃, and inoculating spores concentration is 10
11The Rhizopus oryzae of individual/L (Rhizopus oryzae) AS3.819 spore suspension 500mL carries out aeration-agitation fermentation, fermentation time 60h at 32 ℃-34 ℃; Stop to ventilate and stirring, remaining sugar concentration leaves standstill less than 5g/L in jar; Rhizopus oryzae thalline natural subsidence arrives at the bottom of the fermentor tank, adopts malleation to discharge the L-lactic acid fermentation liquid of 5/6 volume;
(3) set by step (2) add material again in fermentor tank, carry out the aeration-agitation fermentation at 32 ℃-34 ℃, repeat 25 batches, each batch fermentation time 20h; Each batch fermentation ends stops to ventilate and stirs, recycling at the bottom of the Rhizopus oryzae thalline is stayed jar, and malleation is emitted the L-lactic acid fermentation liquid of 5/6 volume;
Other operation steps is with embodiment 1.
Embodiment 3:
(1) rice straw is pulverized the back and cross 40 mesh sieves, ratio in 4.5L/Kg adds calcium hydroxide aqueous solution, behind 95 ℃ of following heated and stirred 30min, add in 98% vitriol oil extremely neutral with calcium hydroxide, separate solid composition and calcium sulfate precipitation, add enzyme in the ratio of 1.2g/L and live and be the liquid cellulase of 15U/g, 60 ℃ of stirring enzymolysis 70min down make that to contain saccharic amount per-cent be 12% saccharification liquid 10L;
(2) be that 12% saccharification liquid adds in the fermentor tank with the mass percent that makes, add lime carbonate 600g, ammonium nitrate 35g, potassium primary phosphate 5g, SODIUM PHOSPHATE, MONOBASIC 3g, zinc sulfate 4g, sal epsom 4g and ferrous sulfate 1.0g simultaneously, sterilization is cooled to 32 ℃-34 ℃, and inoculating spores concentration is 10
11The Rhizopus oryzae of individual/L (Rhizopus oryzae) AS3.819 spore suspension 500mL carries out aeration-agitation fermentation, fermentation time 65h at 32 ℃-34 ℃; Stop to ventilate and stirring, remaining sugar concentration leaves standstill less than 5g/L in jar; Rhizopus oryzae thalline natural subsidence arrives at the bottom of the fermentor tank, adopts malleation to discharge the L-lactic acid fermentation liquid of 5/6 volume;
(3) set by step (2) add material again in fermentor tank, carry out the aeration-agitation fermentation at 32 ℃-34 ℃, repeat 25 batches, each batch fermentation time 22h; Each batch fermentation ends stops to ventilate and stirs, recycling at the bottom of the Rhizopus oryzae thalline is stayed jar, and malleation is emitted the L-lactic acid fermentation liquid of 5/6 volume; Other operation steps is with embodiment 1.
Claims (2)
1. method of producing L-lactic acid is characterized in that comprising following operation steps:
(1) agricultural crop straw is pulverized the back and cross 40 mesh sieves, employing sig water, diluted acid, cellulase are handled successively, make to contain the saccharification liquid 10L that saccharic amount per-cent is 8-12%;
(2) be that 8-12% saccharification liquid adds in the fermentor tank with the mass percent that makes, add lime carbonate 400-600g, ammonium nitrate 30-35g, potassium primary phosphate 3-5g, SODIUM PHOSPHATE, MONOBASIC 2-3g, zinc sulfate 3-4g, sal epsom 3-4g and ferrous sulfate 0.8-1.0g simultaneously, sterilization is cooled to 32 ℃～34 ℃, and inoculating spores concentration is 10
11The Rhizopus oryzae of individual/L (Rhizopus oryzae) AS3.819 spore suspension 500mL carries out aeration-agitation fermentation, fermentation time 60-65h at 32 ℃～34 ℃; Stop to ventilate and stirring, remaining sugar concentration leaves standstill less than 5g/L in jar; Rhizopus oryzae thalline natural subsidence arrives at the bottom of the fermentor tank, adopts malleation to discharge the L-lactic acid fermentation liquid of 5/6 volume;
(3) set by step (2) add material again in fermentor tank, carry out the aeration-agitation fermentation at 32 ℃～34 ℃, repeat 25 batches, each batch fermentation time 20～22h; Each batch fermentation ends stops to ventilate and stirs, recycling at the bottom of the Rhizopus oryzae thalline is stayed jar, and malleation is emitted the L-lactic acid fermentation liquid of 5/6 volume;
(4) the L-lactic acid fermentation liquid that step (2) and (3) are obtained merges, than adding 4.5 ‰ (w/v) calcium hydroxide, 7.5 ‰ (w/v) gacs and 3 ‰ (w/v) sal epsom, stir 45min down by mass/volume, leave standstill at 70 ℃, impurity is removed in centrifuging, obtains the L-lactic acid fermentation clear liquid;
(5) adopt 98% vitriol oil that the L-lactic acid fermentation clear liquid is carried out acidifying, staticly settle, remove calcium sulfate, obtain acidifying fermentation liquid;
(6) be 3000 daltonian rolling ultra-filtration membranes with acidifying fermentation liquid by molecular weight cut-off, working pressure 0.8Mpa, concentration ratio are 10, remove macromole impurity such as deproteinize, obtain ultrafiltration and concentration liquid and ultrafiltration and see through liquid, ultrafiltration and concentration liquid filters after handling through charcoal absorption again;
(7) ultrafiltration is seen through liquid and adopt nanofiltration membrane to filter, molecular weight cut-off 150～300 dalton, working pressure 1.0MPa accounts for 95% of feed volume through liquid is long-pending, obtains nanofiltration concentrated solution and nanofiltration and sees through liquid, and the nanofiltration concentrated solution enters the ultra-filtration membrane ultrafiltration;
(8) nanofiltration is seen through liquid and adopt the reverse osmosis pre-concentration, working pressure 4MPa obtains massfraction and is 18% L-lactic acid pre-concentration liquid;
(9) L-lactic acid pre-concentration liquid is removed inorganic salt through storng-acid cation exchange resin, weak base anion-exchange resin successively, obtain desalination L-lactic acid solution;
(10) desalination L-lactic acid solution enters the vacuum distilling unit and concentrates, and obtains the L-lactic product of mass percentage concentration 85%, and quality reaches the GB2023-2003 standard.
2. a kind of method of producing L-lactic acid according to claim 1 is characterized in that: described agricultural crop straw is maize straw or wheat stalk or rice straw.
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| CN115991646A (en) * | 2021-10-18 | 2023-04-21 | 上海凯赛生物技术股份有限公司 | Method for separating and extracting lactic acid, lactic acid product and preparation method thereof |
| CN115991643A (en) * | 2021-10-18 | 2023-04-21 | 上海凯赛生物技术股份有限公司 | Method for separating and extracting lactic acid |
-
2010
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102061266B (en) * | 2010-11-23 | 2012-10-03 | 合肥工业大学 | Method for preparing and regenerating rhizopus oryzae protoplast for producing L-lactic acid at high yield |
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