CN103937846B - A kind of method utilizing cotton cake dregs to prepare Moriamin S - Google Patents
A kind of method utilizing cotton cake dregs to prepare Moriamin S Download PDFInfo
- Publication number
- CN103937846B CN103937846B CN201410101521.6A CN201410101521A CN103937846B CN 103937846 B CN103937846 B CN 103937846B CN 201410101521 A CN201410101521 A CN 201410101521A CN 103937846 B CN103937846 B CN 103937846B
- Authority
- CN
- China
- Prior art keywords
- moriamin
- extraction
- cake dregs
- cotton cake
- technique
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
A kind of method utilizing cotton cake dregs to prepare Moriamin S, this method first carries out mechanical disintegration and pretreatment to cotton cake dregs, inoculate aspergillus niger, aspergillus oryzae, yeast saccharomyces cerevisiae cultivate under aerobic conditions 15 20h form one-level enzymatic hydrolysis system, then subtilis is inoculated respectively, Bacillus licheniformis continue aerobic cultivate 18 32h to form fermented liquid, acid hydrolysis is carried out subsequently to adding food grade concentrated hydrochloric acid in fermented liquid, and filter successively after hydrolysis terminates, the sedimentation of acid adjustment degree, obtain Moriamin S, complexing affinity extraction technology is adopted to refine Moriamin S again, the raffinate aqueous phase produced in treating process is by being mixed with light soy sauce with Moriamin S, the extraction agent that raffinate oil phase then can be used as refining next batch Moriamin S recycles.The present invention not only effectively improves raw material availability, reduces production cost, avoids toxic by-products to produce, and adds value-added content of product.
Description
Technical field
The present invention relates to a kind of is raw material with cotton cake dregs, and multi-cultur es cooperative fermentation prepares the method for Moriamin S, is applicable to improve raw material availability, reduces production cost, avoids toxic by-products to produce, improves value-added content of product.
Background technology
Amino acid and derivative thereof are widely applied in fields such as medicine, food, agricultural and forestry.The common production method of single amino acid has chemical synthesis and fermentation method two kinds.Chemical synthesis complex process, with high costs, and kind is incomplete.Microbe fermentation method productive rate is low, and product Nong Shrink energy consumption is large, and kind is incomplete yet.The preparation of aminoacids complex is raw material mainly with the blood of animal, hair, animal residue and vegetable-protein, is realized by bio-transformation (direct fermentation and enzymatic conversion), decomposition, extracting three kinds of modes.
The by product that cotton cake dregs is cottonseed after oil expression, not only cellulose, its crude protein content also up to 33% ?42%.Compare with plant protein sources such as dregs of rapeseed cake with dregs of beans, cotton cake dregs except Methionin and methionine content lower slightly except, other aminoacids contents are all very close with the former, and the arginine content in cotton cake dregs is even far above the former.In addition, the arginine in cotton cake dregs and Methionin ratio are up to more than 2.7, and be 2 times of normal value, visible, cotton cake dregs is first-class plant protein source.But due to the existence of free gossypol, the utility value of cotton cake dregs is had a greatly reduced quality, and for a long time, cotton cake dregs serves as Fertilizer application more, and cost performance is low.
Chinese patent: application publication number is CN1733925A, date of publication is a kind of method that the patent of invention on February 15th, 2006 discloses waste plant protein and prepares Moriamin S, dregs of beans, rapeseed cake and cottonseed cake are first carried out removal of impurities, pulverize, sieve by the method, with dilute sulphuric acid or dilute hydrochloric acid soak, boiling, adopt bacterium neutral protease to carry out enzymic hydrolysis to it again, then add food grade pure hydrochloric acid and acid hydrolysis is carried out to it.Although this invention is simple to operate, safe and reliable, but still there is following defect:
1, in the process of soaking with dilute sulphuric acid or dilute hydrochloric acid, the portion of cellulose in raw material and protein are hydrolyzed, and its hydrolysate is dissolved in supernatant liquor and is removed, therefore, in raw material, the utilization ratio of Mierocrystalline cellulose and protein is lower, and meanwhile, the process of the supernatant liquor be cut also is a difficult problem;
2, in enzyme hydrolysis step, the use of neutral protease not only increases the cost of whole preparation technology, and its protolysate is limited in one's ability, therefore need in acid hydrolysis step by hydrolyzed solution acid concentration of volume percent be adjusted to 33% ?50%, namely acid mass percent concentration be 12% ?18%, this acidity condition is easy to produce organic poison propylene chlorohydrin, causes finished product to lose practicality.
Summary of the invention
The cotton cake dregs that utilizes the object of the invention is to overcome that the raw material availability existed in prior art is low, production cost is high, the problem of easy generation toxic by-products, provide a kind of and can effectively improve raw material availability, reduce production cost, avoided toxic by-products to produce prepares the method for Moriamin S.
For realizing above object, the invention provides following technical scheme:
Utilize cotton cake dregs to prepare a method for Moriamin S, this preparation method comprises following technique successively:
The pretreatment technology of cotton cake dregs: wait to be hydrolyzed material in 110 ?, 130 DEG C of boiling 15 ?20min to be formed after the quality such as the cotton cake dregs after mechanical disintegration and water being mixed;
Bacterium enzyme hydrolysis process: first will treat that hydrolysis material is cooled to 30 ?36 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), subsequently relative humidity be 80% ?88%, and cultivate under the condition of aerobic 15 ?20h to form one-level enzymatic hydrolysis system, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) continue aerobic cultivate 18 ?32h to form fermented liquid, wherein, described aspergillus niger, aspergillus oryzae, yeast saccharomyces cerevisiae, subtilis, the inoculum size of Bacillus licheniformis be all cotton cake dregs quality 0.2% ?0.5%,
Acid hydrolysis process: first add the food grade concentrated hydrochloric acid that massfraction is 36% in fermented liquid, then be hydrolyzed 10 ?15h in 85 ?95 DEG C, wherein, the interpolation quality of described food grade concentrated hydrochloric acid is wait to be hydrolyzed 0.7 ?0.9 times of quality of material;
In filtration and technique: first adopt 300 order filter clothes to filter to obtain filtrate to the fermented liquid after acid hydrolysis, again with solid sodium carbonate or solid sodium bicarbonate the pH value of filtrate is adjusted to 1.5 ?3.2 to form solid-liquid mixing system, then filter to obtain Moriamin S to this solid-liquid mixing system, now, the preparation of Moriamin S completes.
In described bacterium enzyme hydrolysis process, first in one-level enzymatic hydrolysis system, inoculate subtilis respectively, Bacillus licheniformis aerobic cultivate 8 ?12h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 5.0 ?7.0, and in 50 ?continue at 58 DEG C aerobic cultivate 10 ?20h to form fermented liquid, wherein, the water added is 2.5 ?3.5: 1 with the mass ratio waiting to be hydrolyzed material.
In described filtration and in technique, before employing 300 order filter cloth filters the fermented liquid after acid hydrolysis, first with 100 order double gauzes, coarse filtration is carried out to it.
In described filtration and in technique, before filtering solid-liquid mixing system, prior to being left standstill 1 ?4h under room temperature.
Described preparation method also comprises the process for refining of Moriamin S, after this technique is arranged in filtration and technique;
The process for refining of described Moriamin S comprises the following steps successively:
First time extraction a: extraction agent, Moriamin S equal-volume are poured in extraction equipment and carried out first time extraction after mixing to obtain an extraction phase and an extracting phase, wherein, the composition of an extraction agent and volume ratio thereof are: n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 0.5 ?2;
Second time extraction a: extraction phase, No. two extraction agent equal-volumes are poured in extraction equipment and carried out second time extraction after mixing to obtain refining Moriamin S and No. two extracting phases, now, the process for refining of Moriamin S terminates, wherein, described No. two extraction agents to be concentration be 0.35 ?the aqueous hydrochloric acid of 0.7mol/L.
The extraction time of described first time extraction is more than or equal to 30min, and the extraction time of described second time extraction is more than or equal to 5min.
The process for refining of described Moriamin S also comprises procedure of pH adjustment, before this step is positioned at first time extraction step;
Described procedure of pH adjustment refers to: adopt sodium hydroxide solution or potassium hydroxide solution the pH value of Moriamin S is adjusted to 3.5 ?7.
Described preparation method also comprises the recycling technique of extracting phase, after this technique is positioned at the process for refining of Moriamin S;
The recycling technique of described extracting phase comprises the activation of the recycling of an extracting phase, No. two extracting phases after repeatedly recycling;
The recycling of a described extracting phase refers to: first an extracting phase is mixed to form mixed system with Moriamin S equal-volume, again with concentration be 2.0 ?the sodium hydroxide solution of 3.5mol/L the pH value of this mixed system is adjusted to 6.8 ?7.2, in the mixed system after regulating pH, then add salt and sanitas to obtain light soy sauce;
The activation of described No. two extracting phases after repeatedly recycling refers to: first after repeatedly recycling No. two extracting phase is carried out 2 ?4 times strip, then by No. two extracting phase distilled water washs 2 after stripping ?3 times to obtain an extraction agent, wherein, the strippant used in described reextraction for concentration be 1 ?the aqueous hydrochloric acid of 2mol/L, in each reextraction, the volume ratio of strippant and extract is all 1: 1, the time of each reextraction is more than or equal to 5min, and the time of each washing is more than or equal to 1h.
Compared with prior art, beneficial effect of the present invention is:
1, a kind of method utilizing cotton cake dregs to prepare Moriamin S of the present invention comprises bacterium enzyme hydrolysis process, this technique adopts first inoculated aspergillus niger respectively, aspergillus oryzae, yeast saccharomyces cerevisiae aerobic cultivate 15 ?20h, inoculate subtilis respectively again, Bacillus licheniformis aerobic cultivate 18 ?the method of 32h, not only effectively improve the utilization ratio of raw material, and significantly reduce the acid concentration needed for degradable for cotton cake dregs albumen one-tenth amino acid, avoid and adopt high concentrated acid protolysate to form 3-propylene chlorohydrin, in addition, without the need to additionally adding substrate in whole culturing process, also production cost can be reduced while avoiding by product to be formed.Therefore, the present invention not only increases the utilization ratio of raw material, and avoids the formation of toxic by-products, reduces production cost.
2, a kind of method utilizing cotton cake dregs to prepare Moriamin S of the present invention adopts the Moriamin S of the process for refining of Moriamin S to preparation to carry out deep processing, not only eliminate the reducing sugar in Moriamin S, too much chlorion and most L-glutamic acid and ammonium salt, and ensure that the content of indispensable amino acid in the cotton cake dregs proteolysis stoste after amino acid extracts is higher, can as medicine, the additive of amino acids oral-liquor or other health promoting beverage uses, by the recycling technique of extracting phase using an extracting phase producing in the Moriamin S treating process raw material as the light soy sauce of preparation, through repeatedly recycling No. two extracting phase is carried out activation treatment, it is made to continue to use as an extraction agent, increase substantially value-added content of product.Therefore, the present invention can significantly improve value-added content of product.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated.
Utilize cotton cake dregs to prepare a method for Moriamin S, this preparation method comprises following technique successively:
The pretreatment technology of cotton cake dregs: wait to be hydrolyzed material in 110 ?, 130 DEG C of boiling 15 ?20min to be formed after the quality such as the cotton cake dregs after mechanical disintegration and water being mixed;
Bacterium enzyme hydrolysis process: first will treat that hydrolysis material is cooled to 30 ?36 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), subsequently relative humidity be 80% ?88%, and cultivate under the condition of aerobic 15 ?20h to form one-level enzymatic hydrolysis system, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) continue aerobic cultivate 18 ?32h to form fermented liquid, wherein, described aspergillus niger, aspergillus oryzae, yeast saccharomyces cerevisiae, subtilis, the inoculum size of Bacillus licheniformis be all cotton cake dregs quality 0.2% ?0.5%,
Acid hydrolysis process: first add the food grade concentrated hydrochloric acid that massfraction is 36% in fermented liquid, then be hydrolyzed 10 ?15h in 85 ?95 DEG C, wherein, the interpolation quality of described food grade concentrated hydrochloric acid is wait to be hydrolyzed 0.7 ?0.9 times of quality of material;
In filtration and technique: first adopt 300 order filter clothes to filter to obtain filtrate to the fermented liquid after acid hydrolysis, again with solid sodium carbonate or solid sodium bicarbonate the pH value of filtrate is adjusted to 1.5 ?3.2 to form solid-liquid mixing system, then filter to obtain Moriamin S to this solid-liquid mixing system, now, the preparation of Moriamin S completes.
In described bacterium enzyme hydrolysis process, first in one-level enzymatic hydrolysis system, inoculate subtilis respectively, Bacillus licheniformis aerobic cultivate 8 ?12h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 5.0 ?7.0, and in 50 ?continue at 58 DEG C aerobic cultivate 10 ?20h to form fermented liquid, wherein, the water added is 2.5 ?3.5: 1 with the mass ratio waiting to be hydrolyzed material.
In described filtration and in technique, before employing 300 order filter cloth filters the fermented liquid after acid hydrolysis, first with 100 order double gauzes, coarse filtration is carried out to it.
In described filtration and in technique, before filtering solid-liquid mixing system, prior to being left standstill 1 ?4h under room temperature.
Described preparation method also comprises the process for refining of Moriamin S, after this technique is arranged in filtration and technique;
The process for refining of described Moriamin S comprises the following steps successively:
First time extraction a: extraction agent, Moriamin S equal-volume are poured in extraction equipment and carried out first time extraction after mixing to obtain an extraction phase and an extracting phase, wherein, the composition of an extraction agent and volume ratio thereof are: n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 0.5 ?2;
Second time extraction a: extraction phase, No. two extraction agent equal-volumes are poured in extraction equipment and carried out second time extraction after mixing to obtain refining Moriamin S and No. two extracting phases, now, the process for refining of Moriamin S terminates, wherein, described No. two extraction agents to be concentration be 0.35 ?the aqueous hydrochloric acid of 0.7mol/L.
The extraction time of described first time extraction is more than or equal to 30min, and the extraction time of described second time extraction is more than or equal to 5min.
The process for refining of described Moriamin S also comprises procedure of pH adjustment, before this step is positioned at first time extraction step;
Described procedure of pH adjustment refers to: adopt sodium hydroxide solution or potassium hydroxide solution the pH value of Moriamin S is adjusted to 3.5 ?7.
Described preparation method also comprises the recycling technique of extracting phase, after this technique is positioned at the process for refining of Moriamin S;
The recycling technique of described extracting phase comprises the activation of the recycling of an extracting phase, No. two extracting phases after repeatedly recycling;
The recycling of a described extracting phase refers to: first an extracting phase is mixed to form mixed system with Moriamin S equal-volume, again with concentration be 2.0 ?the sodium hydroxide solution of 3.5mol/L the pH value of this mixed system is adjusted to 6.8 ?7.2, in the mixed system after regulating pH, then add salt and sanitas to obtain light soy sauce;
The activation of described No. two extracting phases after repeatedly recycling refers to: first after repeatedly recycling No. two extracting phase is carried out 2 ?4 times strip, then by No. two extracting phase distilled water washs 2 after stripping ?3 times to obtain an extraction agent, wherein, the strippant used in described reextraction for concentration be 1 ?the aqueous hydrochloric acid of 2mol/L, in each reextraction, the volume ratio of strippant and extract is all 1: 1, the time of each reextraction is more than or equal to 5min, and the time of each washing is more than or equal to 1h.
Principle of the present invention is described as follows:
The present invention take cotton cake dregs as raw material, carries out solid-state cooperative fermentation, on the one hand by multi-cultur es to it, effectively reduce the toxicity of cotton cake dregs, make the robust fibre in cotton cake dregs be converted into can fermented type sugar, become tropina after being utilized by microorganism, improve the content of cotton cake dregs albumen, on the other hand, the mode adopting bacterium enzymic hydrolysis and low acid hydrolysis to combine, avoids the generation using 3-propylene chlorohydrin in high salt concentration acid hydrolysis, and the Moriamin S lighter color after hydrolysis, is conducive to further deep processing, moreover, the Moriamin S that process of the present invention obtains after filtration, the sedimentation of acid adjustment degree, complexing affinity extraction technology is adopted to extract, some amino acid in aqueous phase are made to be transferred to organic phase, extracting phase can prepare light soy sauce after allotment, the amino acid be carried in organic phase gets back to aqueous phase after second time extraction, realize the refining of aminoacids complex, leucine is rich in the refining aminoacids complex obtained, Isoleucine, α-amino-isovaleric acid, phenylalanine and arginine etc., can be used for medicine, food, amino acids oral-liquor or be rich in the additive of aminoacid functional beverage, whole production process environmental protection, three-waste free discharge, production cost is low, significantly improve the added value of cotton cake dregs.
A: the pretreatment technology of cotton cake dregs:
This technique carries out boiling after the cotton cake dregs after mechanical disintegration and water being mixed, make the robust fibre in cotton cake dregs and albumen is swelling, loose, be beneficial to cultivation and the fermentation of microorganism.
B: bacterium enzyme hydrolysis process:
This technique first adopts the method for two step inoculation culture, namely the first step respectively inoculated aspergillus niger, aspergillus oryzae, yeast saccharomyces cerevisiae aerobic cultivate 15 ?20h, second step inoculates subtilis more respectively, Bacillus licheniformis aerobic cultivate 18 ?32h, cotton cake dregs proteolysis is become low-molecular-weight oligopeptides by this method, significantly reducing the later stage becomes amino acid whose sour consumption by cotton cake dregs albumen complete hydrolysis, also reaches the object of detoxification simultaneously;
The object of the first step inoculation be the cotton cake dregs protein delivery that makes to be wrapped up by Mierocrystalline cellulose out, increase proteolytic enzyme and its contact surface, be beneficial to hydrolysis.Under aerobic conditions, aspergillus niger energy eccrine fiber element enzyme, polygalacturonase, glucose oxidase, amylase, the multiple enzyme system such as aspartic protease, the macromolecular cpds such as the Mierocrystalline cellulose in cotton cake dregs are converted into fermentable sugar, a fermentable sugar part is utilized by yeast saccharomyces cerevisiae, be decomposed into carbonic acid gas and water, another part is then as the substratum of fermentable, tropina is converted into after microorganism utilizes, and then improve the content of albumen, aspergillus oryzae is the bacterial classification abounding with proteolytic enzyme, simultaneously also can secreting amylase, saccharifying enzyme, cellulase etc., cottonseed cake proteolytic degradation can not only be peptone by these enzyme systems, polypeptide and oligopeptide, the robust fibre in cotton cake dregs can also be made, phytic acid is degraded, the synergy of these three kinds of bacterium, cotton cake dregs albumen and cellulosic degraded are effectively promoted, meanwhile, in whole culturing process, without the need to additionally adding substrate, avoiding the residual of by product, reducing production cost, in addition, because the long meeting of incubation time easily grows miscellaneous bacteria, cause the color and luster of Moriamin S prepared to deepen, increase follow-up refining burden, therefore the time controling cultivated by aerobic of this technique is at 15 ?20h.
Second step inoculation introduces subtilis, Bacillus licheniformis, these two kinds of bacterium are at a large amount of oxygen of necessary for growth, deprive the oxygen required for the breeding of aspergillus niger, aspergillus oryzae and yeast saccharomyces cerevisiae, the growth of aspergillus niger, aspergillus oryzae and yeast saccharomyces cerevisiae can be suppressed, also the breeding of miscellaneous bacteria can be suppressed, make the lighter color of Moriamin S, be beneficial to purifying; In addition, subtilis, Bacillus licheniformis self can also synthesize the enzymes such as α-amylase, proteolytic enzyme, lipase, cellulase, these multiply anchor-pile make the robust fibre in cottonseed cake albumen be hydrolyzed further, and cotton cake dregs albumen is hydrolyzed into the lower oligopeptide of molecular weight; After a large amount of oxygen is deprived by subtilis and Bacillus licheniformis, yeast saccharomyces cerevisiae can will can be hydrolyzed into carbonic acid gas and alcohol for fermented type sugar under anoxic conditions, thus gives Moriamin S special fragrance; In addition, in order to make the multiply anchor-pile of five kinds of bacterium secretion play vigor to greatest extent, this technique after incubation the phase pH value of secondary enzymatic hydrolysis system is adjusted to 5.0 ?7.0, culture temperature control 50 ?58 DEG C.
C: acid hydrolysis process:
The interpolation quality control of food grade concentrated hydrochloric acid is being waited 0.7 ?0.9 times being hydrolyzed quality of material by this technique, to make in fermented liquid the mass percent concentration of acid maintain 5.5% ?7.5%, the hydrolysis environment of this Low acid not only ensure that cotton cake dregs albumen can thoroughly be hydrolyzed at short notice, effectively can also avoid the formation of by product 3-propylene chlorohydrin.
D: in filtration and technique:
This technique first carries out coarse filtration with 100 order double gauzes to the fermented liquid after acid hydrolysis, then adopts 300 order filter clothes to filter it, obviously can accelerate filtering velocity, and meanwhile, filter cloth can also tackle a part of pigment and small-particle impurity.
Reduce amino acid whose loss while making the pigment sedimentation in the fermented liquid after acid hydrolysis as far as possible, and ensure that in the fermented liquid before and after neutralization after acid hydrolysis, amino acid whose change in concentration is little, also desolventing technology is convenient to, this technique with solid sodium carbonate or solid sodium bicarbonate the pH value of filtrate is adjusted to 1.5 ?3.2, and leave standstill 1 ?after 4h sedimentation more thorough.
E: the process for refining of Moriamin S:
Due to Cl in the hydrolyzed solution after hydrochloric acid hydrolysis
-content is at 5.5%-7.5%, and the content of L-glutamic acid is obviously higher, also containing a certain amount of reducing sugar, NH
4 +with a small amount of pigment.For this reason, although this hydrolyzed solution is abundant Moriamin S, will use as the additive of oral liquid or other nourishing drinks, must make further purification process, therefore, this technique adopts twice extracting operation to refine Moriamin S.
Before carrying out first time extracting operation, for improving extraction efficiency as far as possible, this technique have employed sodium hydroxide solution or potassium hydroxide solution the pH value of Moriamin S is adjusted to 3.5 ?7 method, the use of sodium hydroxide solution or potassium hydroxide solution makes the ionic concn change in Moriamin S little, and the by product of formation is less.
Some important amino acids in Moriamin S (as phenylalanine, methionine(Met), α-amino-isovaleric acid, leucine, Isoleucine, Histidine etc.) proceed in organic phase by solvent extraction by first time extraction, achieve being separated of important amino acid and chlorion, reducing sugar and most ammonium salt.
The amino acid complete back extraction backwater phase that second time extraction adopts the aqueous hydrochloric acid (0.35 ?0.7mol/L) of lower concentration to extract in organic phase, through this operation, in the refining Moriamin S obtained, chloride ion content is low, not containing reducing sugar, content of glutamic acid drops to suitable concentration range, the content of indispensable amino acid is high, total amino acid content is more than 1%, can use as the additive of oral liquid or other nourishing drinks, and produce in this step No. two extracting phases can be used as an extraction agent and repeatedly recycle.
The recycling of f: number extracting phase:
Containing a large amount of amino acid in the extracting phase produced in the process for refining of Moriamin S, wherein content of glutamic acid is the highest, glycine, L-Ala, Threonine, Serine, ornithine, γ-aminobutyric acid etc. also occupy suitable proportion, also containing a certain amount of reducing sugar, if it is directly made soy sauce, essential amino acids content is wherein on the low side, and therefore, itself and Moriamin S are mixed with light soy sauce by this technique.
The activation of g: No. two extracting phases:
After repeatedly recycling No. two extracting phase is carried out activation treatment by this technique, makes it continue to use as an extraction agent.In order to remove the NH be carried in No. two extracting phases
4 +and pigment, this process selection concentration be 1 ?the aqueous hydrochloric acid of 2mol/L as strippant, it is repeatedly stripped, and the use of aqueous hydrochloric acid will inevitably cause load in No. two extracting phases after stripping to have more hydrogen ion, therefore adopt distilled water repeatedly to wash and be removed.
Embodiment 1:
Utilize cotton cake dregs to prepare a method for Moriamin S, this preparation method comprises following technique successively:
The pretreatment technology of cotton cake dregs: wait to be hydrolyzed material in 121 DEG C of boiling 20min to be formed after the quality such as the cotton cake dregs after mechanical disintegration and water being mixed;
Bacterium enzyme hydrolysis process: first will treat that hydrolysis material is cooled to 30 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), be 80% in relative humidity subsequently, and aerobic cultivates 15h to form one-level enzymatic hydrolysis system under the condition of aerobic, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) aerobic cultivates 8h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 6.5, and at 55 DEG C, continue aerobic cultivation 12h to form fermented liquid, wherein, described aspergillus niger, aspergillus oryzae, yeast saccharomyces cerevisiae, subtilis, the inoculum size of Bacillus licheniformis is all 0.4% of cotton cake dregs quality, the water added is 3: 1 with the mass ratio waiting to be hydrolyzed material,
Acid hydrolysis process: first add the food grade concentrated hydrochloric acid that massfraction is 36% in fermented liquid, then in 90 DEG C of hydrolysis 12h, wherein, the interpolation quality of described food grade concentrated hydrochloric acid is 0.8 times that waits to be hydrolyzed quality of material;
In filtration and technique: first by the fermented liquid after acid hydrolysis successively through 100 order double gauzes, 300 order filter-cloth filterings to obtain filtrate, with solid sodium carbonate, the pH value of filtrate is adjusted to 1.5 to form solid-liquid mixing system again, then this solid-liquid mixture is lain in left at room temperature 1h, filter to obtain Moriamin S to it again, now, the preparation of Moriamin S completes.
The process for refining of Moriamin S:
A:pH regulates: employing concentration is that the pH value of Moriamin S is adjusted to 5.0 by the sodium hydroxide solution of 3mol/L;
B: first time extraction a: extraction agent, Moriamin S equal-volume are poured in extraction equipment and carried out first time extraction after mixing to obtain an extraction phase and an extracting phase, wherein, the composition of an extraction agent and volume ratio thereof are: n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 1, and extraction time is 60min;
C: second time extraction a: extraction phase, No. two extraction agent equal-volumes are poured in extraction equipment and carried out second time extraction after mixing to obtain refining Moriamin S and No. two extracting phases, now, the process for refining of Moriamin S terminates, wherein, the aqueous hydrochloric acid of described No. two extraction agents to be concentration be 0.5mol/L, extraction time is 10min.
The recycling technique of extracting phase:
The recycling of a: number extracting phase a: first extracting phase is mixed to form mixed system with Moriamin S equal-volume, be that the pH value of this mixed system is adjusted to 7.0 by the sodium hydroxide solution of 3mol/L again by concentration, in the mixed system after regulating pH, then add salt and sanitas to obtain light soy sauce;
B: the activation of No. two extracting phases after repeatedly recycling: No. two extracting phases after repeatedly recycling carry out 4 times and strip, then by No. two extracting phase distilled water washs 3 times after stripping to obtain an extraction agent, wherein, the strippant used in described reextraction for concentration be the aqueous hydrochloric acid of 2mol/L, in each reextraction, the volume ratio of strippant and extract is all 1: 1, the time of each reextraction is 10min, and the time of each washing is 1h.
Embodiment 2:
Step with embodiment 1, unlike:
In described bacterium enzyme hydrolysis process, first will treat that hydrolysis material is cooled to 34 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), be 85% in relative humidity subsequently, and aerobic cultivates 20h to form one-level enzymatic hydrolysis system under the condition of aerobic, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) aerobic cultivates 12h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 5.0, and at 50 DEG C, continue aerobic cultivation 15h to form fermented liquid, wherein, described aspergillus niger, aspergillus oryzae, the inoculum size of yeast saccharomyces cerevisiae is all 0.3% of cotton cake dregs quality, the water added is 3.5: 1 with the mass ratio waiting to be hydrolyzed material,
In described acid hydrolysis process, first in fermented liquid, add food grade concentrated hydrochloric acid, then in 95 DEG C of hydrolysis 10h, wherein, the interpolation quality of described food grade concentrated hydrochloric acid is 0.9 times that waits to be hydrolyzed quality of material;
In described filtration and in technique, with solid sodium carbonate, the pH value of filtrate is adjusted to 2.5 to form solid-liquid mixing system;
In the process for refining of described Moriamin S, the pH value of Moriamin S is adjusted to 3.5 by procedure of pH adjustment, the composition of an extraction agent and volume ratio thereof are: the aqueous hydrochloric acid of n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 2, No. two extraction agents to be concentration be 0.35mol/L;
In the recycling technique of described extracting phase, adopt concentration to be that the pH value of this mixed system is adjusted to 6.8 by the sodium hydroxide solution of 3mol/L, and strippant is concentration is the aqueous hydrochloric acid of 2mol/L.
Embodiment 3:
Step with embodiment 1, unlike:
In described bacterium enzyme hydrolysis process, first will treat that hydrolysis material is cooled to 36 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), be 88% in relative humidity subsequently, and aerobic cultivates 20h to form one-level enzymatic hydrolysis system under the condition of aerobic, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) aerobic cultivates 10h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 6.0, and at 53 DEG C, continue aerobic cultivation 22h to form fermented liquid,
In described acid hydrolysis process, first in fermented liquid, add food grade concentrated hydrochloric acid, then in 85 DEG C of hydrolysis 15h, wherein, the interpolation quality of food grade concentrated hydrochloric acid is 0.85 times that waits to be hydrolyzed quality of material;
In the process for refining of described Moriamin S, the pH value of Moriamin S is adjusted to 5.5 by procedure of pH adjustment, the composition of an extraction agent and volume ratio thereof are: the aqueous hydrochloric acid of n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 1.5, No. two extraction agents to be concentration be 0.7mol/L;
In the recycling technique of described extracting phase, employing concentration is that the pH value of this mixed system is adjusted to 7.2 by the sodium hydroxide solution of 3mol/L.
Embodiment 4:
Step with embodiment 1, unlike:
In described bacterium enzyme hydrolysis process, first will treat that hydrolysis material is cooled to 32 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), be 85% in relative humidity subsequently, and aerobic cultivates 20h to form one-level enzymatic hydrolysis system under the condition of aerobic, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) aerobic cultivates 12h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 7.0, and at 58 DEG C, continue aerobic cultivation 18h to form fermented liquid, wherein, described aspergillus oryzae, the inoculum size of yeast saccharomyces cerevisiae is all 0.2% of cotton cake dregs quality, subtilis, the inoculum size of Bacillus licheniformis is all 0.5% of cotton cake dregs quality, the water added is 2.5: 1 with the mass ratio waiting to be hydrolyzed material,
In described acid hydrolysis process, the interpolation quality of food grade concentrated hydrochloric acid is 0.7 times that waits to be hydrolyzed quality of material;
In described filtration and in technique, with solid sodium carbonate, the pH value of filtrate is adjusted to 3.2 to form solid-liquid mixing system;
In the process for refining of described Moriamin S, the pH value of Moriamin S is adjusted to 7.0 by procedure of pH adjustment, and the composition of an extraction agent and volume ratio thereof are: n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 0.5;
In the recycling technique of described extracting phase, employing concentration is that the pH value of this mixed system is adjusted to 6.8 by the sodium hydroxide solution of 3mol/L.
The aminoacids content of the Moriamin S adopting above-mentioned four embodiments to prepare is more than 1%, not containing propylene chlorohydrin, gossypol and reducing sugar, the nitrogen content of ammonium salt is lower than 0.04%, aminoacids content in light color soy sauce is not less than 2.5%, No. two extracting phases after activated are continued use 10 times as an extraction agent, and its extraction ability has no decay.
Claims (8)
1. utilize cotton cake dregs to prepare a method for Moriamin S, it is characterized in that described preparation method comprises following technique successively:
The pretreatment technology of cotton cake dregs: wait to be hydrolyzed material in 110 ?, 130 DEG C of boiling 15 ?20min to be formed after the quality such as the cotton cake dregs after mechanical disintegration and water being mixed;
Bacterium enzyme hydrolysis process: first will treat that hydrolysis material is cooled to 30 ?36 DEG C, difference inoculated aspergillus niger (Aspergillusniger) at this temperature again, aspergillus oryzae (Aspergillusoryzae), yeast saccharomyces cerevisiae (Saccharomycescerevisiae), subsequently relative humidity be 80% ?88%, and cultivate under the condition of aerobic 15 ?20h to form one-level enzymatic hydrolysis system, then in one-level enzymatic hydrolysis system, subtilis (Bacillussubtilis) is inoculated respectively, Bacillus licheniformis (Bacilluslicheniformis) continue aerobic cultivate 18 ?32h to form fermented liquid, wherein, described aspergillus niger, aspergillus oryzae, yeast saccharomyces cerevisiae, subtilis, the inoculum size of Bacillus licheniformis be all cotton cake dregs quality 0.2% ?0.5%,
Acid hydrolysis process: first add the food grade concentrated hydrochloric acid that massfraction is 36% in fermented liquid, then be hydrolyzed 10 ?15h in 85 ?95 DEG C, wherein, the interpolation quality of described food grade concentrated hydrochloric acid is wait to be hydrolyzed 0.7 ?0.9 times of quality of material;
In filtration and technique: first adopt 300 order filter clothes to filter to obtain filtrate to the fermented liquid after acid hydrolysis, again with solid sodium carbonate or solid sodium bicarbonate the pH value of filtrate is adjusted to 1.5 ?3.2 to form solid-liquid mixing system, then filter to obtain Moriamin S to this solid-liquid mixing system, now, the preparation of Moriamin S completes.
2. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 1, it is characterized in that: in described bacterium enzyme hydrolysis process, first in one-level enzymatic hydrolysis system, inoculate subtilis respectively, Bacillus licheniformis aerobic cultivate 8 ?12h to form secondary enzymatic hydrolysis system, water is added again in secondary enzymatic hydrolysis system, then the pH value adding the secondary enzymatic hydrolysis system after water is adjusted to 5.0 ?7.0, and in 50 ?continue at 58 DEG C aerobic cultivate 10 ?20h to form fermented liquid, wherein, the water added is 2.5 ?3.5: 1 with the mass ratio waiting to be hydrolyzed material.
3. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 1 and 2, it is characterized in that: in described filtration and in technique, before employing 300 order filter cloth filters the fermented liquid after acid hydrolysis, first with 100 order double gauzes, coarse filtration is carried out to it.
4. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 1 and 2, is characterized in that: in described filtration and in technique, before filtering solid-liquid mixing system, prior to being left standstill 1 ?4h under room temperature.
5. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 1 and 2, is characterized in that:
Described preparation method also comprises the process for refining of Moriamin S, after this technique is arranged in filtration and technique;
The process for refining of described Moriamin S comprises the following steps successively:
First time extraction a: extraction agent, Moriamin S equal-volume are poured in extraction equipment and carried out first time extraction after mixing to obtain an extraction phase and an extracting phase, wherein, the composition of an extraction agent and volume ratio thereof are: n-Octanol 1, di-(2-ethylhexyl)phosphoric acid ester 0.5 ?2;
Second time extraction a: extraction phase, No. two extraction agent equal-volumes are poured in extraction equipment and carried out second time extraction after mixing to obtain refining Moriamin S and No. two extracting phases, now, the process for refining of Moriamin S terminates, wherein, described No. two extraction agents to be concentration be 0.35 ?the aqueous hydrochloric acid of 0.7mol/L.
6. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 5, is characterized in that: the extraction time of described first time extraction is more than or equal to 30min, and the extraction time of described second time extraction is more than or equal to 5min.
7. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 5, is characterized in that:
The process for refining of described Moriamin S also comprises procedure of pH adjustment, before this step is positioned at first time extraction step;
Described procedure of pH adjustment refers to: adopt sodium hydroxide solution or potassium hydroxide solution the pH value of Moriamin S is adjusted to 3.5 ?7.
8. a kind of method utilizing cotton cake dregs to prepare Moriamin S according to claim 5, is characterized in that:
Described preparation method also comprises the recycling technique of extracting phase, after this technique is positioned at the process for refining of Moriamin S;
The recycling technique of described extracting phase comprises the activation of the recycling of an extracting phase, No. two extracting phases after repeatedly recycling;
The recycling of a described extracting phase refers to: first an extracting phase is mixed to form mixed system with Moriamin S equal-volume, again with concentration be 2.0 ?the sodium hydroxide solution of 3.5mol/L the pH value of this mixed system is adjusted to 6.8 ?7.2, in the mixed system after regulating pH, then add salt and sanitas to obtain light soy sauce;
The activation of described No. two extracting phases after repeatedly recycling refers to: first after repeatedly recycling No. two extracting phase is carried out 2 ?4 times strip, then by No. two extracting phase distilled water washs 2 after stripping ?3 times to obtain an extraction agent, wherein, the strippant used in described reextraction for concentration be 1 ?the aqueous hydrochloric acid of 2mol/L, in each reextraction, the volume ratio of strippant and extract is all 1: 1, the time of each reextraction is more than or equal to 5min, and the time of each washing is more than or equal to 1h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410101521.6A CN103937846B (en) | 2014-03-19 | 2014-03-19 | A kind of method utilizing cotton cake dregs to prepare Moriamin S |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410101521.6A CN103937846B (en) | 2014-03-19 | 2014-03-19 | A kind of method utilizing cotton cake dregs to prepare Moriamin S |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103937846A CN103937846A (en) | 2014-07-23 |
| CN103937846B true CN103937846B (en) | 2016-03-09 |
Family
ID=51185715
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410101521.6A Expired - Fee Related CN103937846B (en) | 2014-03-19 | 2014-03-19 | A kind of method utilizing cotton cake dregs to prepare Moriamin S |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103937846B (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104222528A (en) * | 2014-10-09 | 2014-12-24 | 张义盼 | Compound amino acid feed additive and preparation method thereof |
| CN104987268A (en) * | 2015-07-19 | 2015-10-21 | 成都市朝森有机肥有限公司 | Selenium-rich amino acid-microbe organic fertilizer |
| CN105175161A (en) * | 2015-10-22 | 2015-12-23 | 浙江省农业科学院 | Composite amino acid liquid fertilizer and preparation method thereof |
| CN105884405A (en) * | 2015-11-03 | 2016-08-24 | 鹤壁市禾盛生物科技有限公司 | Preparation method of amino acid-polypeptide liquid fertilizer |
| CN105461388A (en) * | 2015-12-08 | 2016-04-06 | 石河子大学 | Amino acid foliar fertilizer prepared from cottonseed meal and preparation method of amino acid foliar fertilizer |
| CN105779543A (en) * | 2016-04-13 | 2016-07-20 | 武汉轻工大学 | Method for preparing organic rapeseed protein through enzyme method |
| CN109170793A (en) * | 2018-09-04 | 2019-01-11 | 上海爱普食品工业有限公司 | A method of acid hydrolyzed vegetable protein baste is produced using pre fermentation dregs of beans |
| CN109517867B (en) * | 2018-11-26 | 2021-10-15 | 中国农业科学院油料作物研究所 | Application of extreme acid and alkali hydrolysis method in production of iturin A by bacillus fermented oil meal and method thereof |
| CN110089615A (en) * | 2019-05-17 | 2019-08-06 | 南通普悦生物医药有限公司 | The preparation method of compound amino acid |
| CN113151032A (en) * | 2020-12-17 | 2021-07-23 | 新希望六和股份有限公司 | Bacillus subtilis with efficient gossypol degradation capability and application thereof |
| CN114516772A (en) * | 2021-08-30 | 2022-05-20 | 鹤壁市禾盛生物科技有限公司 | Rapid fermentation preparation method of high-nitrogen organic liquid fertilizer |
| CN113819752B (en) * | 2021-10-09 | 2024-06-11 | 沂南县坤鹏饲料助剂有限公司 | Animal residue processing system and animal residue processing method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733925A (en) * | 2004-08-12 | 2006-02-15 | 甘肃省科学院生物研究所 | Method with discarded protein Preparation Moriamin S |
| CN102943097A (en) * | 2012-11-15 | 2013-02-27 | 塔城市星河生物工程有限责任公司 | Plant compound amino acid and preparation method as well as application thereof |
-
2014
- 2014-03-19 CN CN201410101521.6A patent/CN103937846B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733925A (en) * | 2004-08-12 | 2006-02-15 | 甘肃省科学院生物研究所 | Method with discarded protein Preparation Moriamin S |
| CN102943097A (en) * | 2012-11-15 | 2013-02-27 | 塔城市星河生物工程有限责任公司 | Plant compound amino acid and preparation method as well as application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 复合微生物固态发酵棉籽饼粕的研究;吴伟伟;《中国优秀硕士学位论文全文数据库农业科技辑》;20100515(第05期);全文 * |
| 菜籽饼的发酵脱毒及酶酸法水解制备复合氨基酸工艺研究;李燕等;《饲料工业》;20111231;第32卷(第8期);51-55 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103937846A (en) | 2014-07-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103937846B (en) | A kind of method utilizing cotton cake dregs to prepare Moriamin S | |
| CN102318724B (en) | Production method of acid soybean peptide protein | |
| CN101979627A (en) | Method for preparing glutamic acid fermentation organic nitrogen additive from glutamic acid fermentation waste thalli | |
| CN110684675B (en) | Blakeslea trispora fermentation method and product thereof | |
| CN105624250A (en) | Enzymolysis-fermentation coupled aquatic protein active peptide preparation method | |
| CN101423855B (en) | Method for preparing polysaccharide by using lucidum strain fermented laminaria leftover | |
| CN106834368A (en) | A kind of method that utilization lignocellulose for fermentation produces L lactic acid | |
| CN102028091A (en) | Method for preparing low-molecular fish peptide by bacillus natto fermentation method | |
| CN110547458A (en) | Preparation method of enzymolysis pearl powder | |
| CN103045705A (en) | Method for producing protein peptide composed of fish protein peptide and soybean peptide | |
| CN103704830A (en) | Method for preparing low-molecule ginsenoside peptide by bacillus natto fermentation | |
| CN101434982B (en) | Method for preparing vegetable seed active peptide by microbial solid state fermentation | |
| CN101434981B (en) | Method for preparing vegetable seed peptide with single bioactivity by microbial solid state fermentation | |
| CN114015739A (en) | Method for preparing liquid collagen peptide from tilapia skin | |
| CN104357428A (en) | Liquid submerged fermentation method of xylanase | |
| CN103725739A (en) | Method for preparing low-molecule sea cucumber peptide by using bacillus natto fermentation method | |
| CN103864954A (en) | Method for extracting peanut meal polysaccharide | |
| CN101654697B (en) | Method for preparing rapeseed peptides by mixed fermentation | |
| CN112708645A (en) | Method for efficiently producing monosodium glutamate | |
| CN104800110B (en) | A kind of method that microbial fermentation prepares total saponin of sapindusmukerossi | |
| CN102524742B (en) | Preparation method of secondary-fermented soy sauce | |
| CN101492708B (en) | Method for preparing vegetable seed peptide with single bioactivity by mix bacterium solid state fermentation | |
| CN101440390B (en) | Method for preparing vegetable seed active peptide by mixed bacteria solid-state fermentation | |
| CN101654693B (en) | Method for preparing rapeseed peptides by microorganism fermentation | |
| CN105624242A (en) | Method for preparing active peptide through high-temperature fermentation of aquatic protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160309 Termination date: 20180319 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |