CN102618610B - Preparation method of sericin compound amino acid - Google Patents
Preparation method of sericin compound amino acid Download PDFInfo
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- CN102618610B CN102618610B CN2012100820181A CN201210082018A CN102618610B CN 102618610 B CN102618610 B CN 102618610B CN 2012100820181 A CN2012100820181 A CN 2012100820181A CN 201210082018 A CN201210082018 A CN 201210082018A CN 102618610 B CN102618610 B CN 102618610B
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Abstract
The invention discloses a preparation method of sericin compound amino acid. The preparation method comprises the following steps of: (1) taking husks and/or frisons as raw materials, and degumming after cleaning; (2) recovering degumming liquid, obtaining sericin solution and adding 0.4-0.6% of preservative into the sericin solution; (3) carrying out enzymolysis, to be specific, firstly adjusting the pH value of the sericin solution obtained in the step (2) to be 9-11, and then hydrolyzing the sericin solution by using 2709 alkaline protease; (4) carrying out acidolysis; (5) carrying out neutralization; (6) carrying out filter pressing and decoloration; (7) treating the solution by adopting macroporous weak-acid acrylic cation exchange resin, macroporous weak-alkaline styrene anion exchange resin and cation-anion mixed type exchange resin in sequence; and (8) carrying out vacuum concentration and spray drying, and obtaining the sericin compound amino acid. Experiments show that the sericin compound amino acid obtained by adopting the method is rich in essential amino acids of a human body and a certain content of tryptophan, solves the problem that the tryptophan can not be obtained in the prior art and has the positive significance for developing nutritional and health-care foods.
Description
Technical field
The present invention relates to a kind of preparation method of silk gum aminoacids complex.
Background technology
Silk is comprised of internal layer fibroin and outer silk gum two portions.Silk gum is coated on the fibroin periphery, and fibroin is played to protection and gluing effect, can prevent because weaving friction to the damage of silky fibre, so silk fabrics all will be to dyeing, ability all purifies silk gum during finishing process.Silk is practiced in the cocoon cooking of the concise liquid sewage of dye-works and silk mill and sewage that the filature workshop gives off, all contains the sericin of very high concentrations, and directly discharge can cause environmental pollution.Silk gum solution after natural silk degumming through purifying, decolouring, degraded, concentrated, spraying drying can make the processable sericin powder, both effectively prevented the pollution of silk gum sewage to environment, greatly improved again the utility value of silk gum.The molecular conformation of processable sericin powder be take the random coil structure as main, and the aggregated structure of superpolymer is mainly amorphous structure.After being widely used in health textile, this novel silk byproduct of silk gum arranges in the fields such as coated material, cosmetics additive, protective foods, medicine, functional living being material.The new purposes of developing silk gum is to be rich in from now on the applicability problem of significance of times in silk byproduct comprehensive utilization field.
Research is found, contain tens seed amino acids in silk gum, wherein the content of eight seed amino acids of needed by human is up to 20%, and this can't obtain so high content in normal food, and the contained mutual ratio of essential amino acid of silk gum is suitable, be to supplement the high-quality source of nutrition that human body needs.
At present, the preparation method for the aminoacids complex in silk gum there is no report.Chinese invention patent CN1313015C discloses a kind of preparation method of powder for refining silk, and it is the product after the silk fibroin hydrolysis, specifically comprises the steps: that (1) is by silkworm cocoon or waste cocoon degumming of silk; (2) adopt 20 ~ 30% sulfuric acid that degumed silk is carried out to acidolysis; (3) add the alkali neutralization; (4) press filtration, decolouring; (5) vacuum concentration, spraying drying, obtain pulverous powder for refining silk.
Yet, the experiment discovery, while adopting above-mentioned preparation method to process sericin, tryptophane wherein can suffer huge destruction, causes in product basically not existing tryptophane.Tryptophane is a kind of indispensable amino acid of human body, and human body can not synthesize, and it must draw from food, and it is the thrombotonin precursor of (also claiming " serotonin "), and thrombotonin is important neurotransmitter.
Therefore, develop a kind of preparation method of silk gum aminoacids complex, to enrich the amino acid of needed by human, especially tryptophane, there is positive realistic meaning.
Summary of the invention
The object of the invention is to provide a kind of preparation method of silk gum aminoacids complex.
For achieving the above object, the technical solution used in the present invention is: a kind of preparation method of silk gum aminoacids complex comprises the steps:
(1) take husks and/or long telling as raw material, come unstuck after cleaning;
(2) reclaim come unglued liquid, husks and/or length are told to dehydration, then use deionized water wash, dehydration liquid, washings and come unglued liquid are merged, obtain silk gum solution, add 0.4 ~ 0.6% sanitas in silk gum solution;
(3) enzymolysis: first the pH value of upper step silk gum solution is adjusted into to 9 ~ 11, then use the above-mentioned silk gum solution of 2709 hydrolysis by novo, temperature is 40 ~ 50 ℃, enzyme concn is 750 ~ 800 unit of activity/g, reaction times 4 ~ 10h is total free aminoacids and part of polypeptide until detected result is major part;
(4) acidolysis: add 10 ~ 15% sulphuric acid hydrolysis in above-mentioned silk gum solution, micro-boiling reflux, be hydrolyzed 10 ~ 15h, until detected result is the overwhelming majority, is total free aminoacids and contains a small amount of dipeptides, tripeptides;
(5) neutralization: add CaO in the silk gum acid hydrolysis liquid obtained in upper step, the pH value of solution is adjusted to 1.5 ~ 2.5, then adding hydrated barta to be neutralized to the pH value is 7.0;
(6) press filtration, decolouring;
(7) adopt successively macropore acidulous acrylic acid cation exchange resin, macroreticular weakly base styrene series anion exchange resin, zwitterion mixed type exchange resin to process upper step solution;
(8) vacuum concentration, spraying drying, can obtain described silk gum aminoacids complex.
Above, in described step (2), add 0.4 ~ 0.6% sanitas in silk gum solution, refer to mass ratio.Described sanitas can be sanitas CY-1 commonly used.
The present invention first utilizes enzyme to open peptide bond to the height specificity of Protein peptide bond, makes the peptide bond partial hydrolysis of sericine molecule, and then makes it thoroughly to be hydrolyzed into total free aminoacids with acid hydrolyzation.
The molecular weight of the silk gum in described silk gum solution is 10000 ~ 150000D.
In technique scheme, in described step (1), the bath raio of coming unstuck is 1:15 ~ 20, and pressure is 0.15 ~ 0.3MPa, and 100 ~ 140min comes unstuck under 120 ~ 140 ℃.
In technique scheme, in described step (3), first with sodium carbonate, the pH value of upper step silk gum solution is adjusted into to 9 ~ 11.
The present invention asks for protection the silk gum aminoacids complex obtained by above-mentioned preparation method simultaneously.
Because technique scheme is used, the present invention compared with prior art has following advantages:
1. the present invention has developed a kind of preparation method of silk gum aminoacids complex, take husks and/or long telling as raw material, prepared the silk gum aminoacids complex in its come unglued liquid, experiment shows, adopt the silk gum aminoacids complex that method of the present invention obtains can enrich essential amino acid, especially there is the tryptophane of certain content, solved and can't obtain the problem of tryptophane in the prior art, there is positive meaning.
2. the present invention first carries out enzymolysis, and then carries out acidolysis, and the concentration of the sulfuric acid adopted in acidolysis is not high, and hydrolysis time is not long yet, thereby its destruction for tryptophane is far smaller than method of the prior art; Simultaneously, the concentration of not high sulfuric acid has also been saved medicine for follow-up neutralization procedure, has simplified operation, thereby has greatly saved cost.
3. simple, the easy handling of preparation method of the present invention, but and save energy, the exploitation nutritive health-care food is had a good application prospect.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment mono-
A kind of preparation method of silk gum aminoacids complex, comprise the steps:
(1) take the 100g husks, clean to clean with tap water, then wash 2 ~ 3 times with deionized water, and use centrifuge dewatering;
(2) come unstuck: it is 1:15 that husks is made into to bath raio, then puts into the pressure cooker 120min that comes unstuck under 125 ℃;
(3) come unglued liquid is reclaimed, and husks is dewatered, then use deionized water wash, dehydration liquid, washings and come unglued liquid are merged, obtain 2000mL silk gum solution, add 0.5% sanitas CY-1 in silk gum solution;
(4) mensuration of solid content: get 20mL silk gum solution and claim it and the total mass of beaker, beaker is put into to baking oven, till evaporating fully to moisture, claim residual mass, both differences are solid content, and the solid content of the present embodiment is every 20mL solution 0.4g;
(5) enzymolysis: first with sodium carbonate, the pH value of upper step silk gum solution is adjusted into to 10, then use the above-mentioned silk gum solution of 2709 hydrolysis by novo, temperature is 45 ℃, enzyme concn is 800 unit of activity/g, reaction times 7h, each hour checked hydrolysis degree with biuret reagent, and each hour 20mL that keeps sample, until detected result is major part, is total free aminoacids and part of polypeptide; When enzymolysis stops, biuret reagent check silk gum hydrolyzed solution is lilac red;
(6) acidolysis: add 12% sulphuric acid hydrolysis in above-mentioned silk gum solution, micro-boiling reflux, with the hydrolysis degree of biuret reagent and ninhydrin reagent check silk gum, from color reaction, after hydrolysis 14h, Proteins In Aqueous Solutions is hydrolyzed substantially; If sericin is hydrolysis thoroughly, just biuret reaction can not occur, and while checking with ninhydrin reagent, generate the bluish voilet product;
(7) neutralization: add CaO in the silk gum acid hydrolysis liquid obtained in upper step, the pH value of solution is adjusted to 2.0, then adding hydrated barta to be neutralized to the pH value is 7.0;
(8) press filtration, decolouring: the neutralizer press filtration is removed to precipitation, and wash precipitation with clear water, add activated carbon decolorizing 1h in clear liquid, 60 ℃ of bleaching temperatures, the gac add-on is destainer 5%, then carries out the press filtration separation, removes filter residue, gets it and crosses cleaner liquid;
(9) filtrate purifying: adopt successively macropore acidulous acrylic acid cation exchange resin, macroreticular weakly base styrene series anion exchange resin, zwitterion mixed type exchange resin to process upper step solution;
(10) vacuum concentration, spraying drying, can obtain described silk gum aminoacids complex.
Adopt the L-8800 of Hitachi high-speed amino acid analyzer to be detected aminoacids complex obtained above, result is as follows:
From table, contain a certain amount of tryptophane in the silk gum aminoacids complex that the present embodiment prepares, be about 0.51%, solved and can't obtain the problem of tryptophane in the prior art, there is positive meaning.
Embodiment bis-
A kind of preparation method of silk gum aminoacids complex, comprise the steps:
(1) take 150g length and tell, clean to clean with tap water, then wash 2 ~ 3 times with deionized water, and use centrifuge dewatering;
(2) come unstuck: it is 1:20 that husks is made into to bath raio, then puts into the pressure cooker 120min that comes unstuck under 125 ℃;
(3) come unglued liquid is reclaimed, and husks is dewatered, then use deionized water wash, dehydration liquid, washings and come unglued liquid are merged, obtain 4000mL silk gum solution, add 0.5% sanitas CY-1 in silk gum solution;
(4) mensuration of solid content: get 20mL silk gum solution and claim it and the total mass of beaker, beaker is put into to baking oven, till evaporating fully to moisture, claim residual mass, both differences are solid content, and the solid content of the present embodiment is every 20mL solution 0.3g;
(5) enzymolysis: first with sodium carbonate, the pH value of upper step silk gum solution is adjusted into to 10, then use the above-mentioned silk gum solution of 2709 hydrolysis by novo, temperature is 45 ℃, enzyme concn is 800 unit of activity/g, reaction times 8h, each hour checked hydrolysis degree with biuret reagent, and each hour 20mL that keeps sample, until detected result is the overwhelming majority, is total free aminoacids and part of polypeptide; When enzymolysis stops, biuret reagent check silk gum hydrolyzed solution is lilac red;
(6) acidolysis: add 12% sulphuric acid hydrolysis in above-mentioned silk gum solution, micro-boiling reflux, with the hydrolysis degree of biuret reagent and ninhydrin reagent check silk gum, from color reaction, after hydrolysis 15h, in solution, sericine is hydrolyzed substantially; If sericin is hydrolysis thoroughly, just biuret reaction can not occur, and while checking with ninhydrin reagent, generate the bluish voilet product;
(7) neutralization: add CaO in the silk gum acid hydrolysis liquid obtained in upper step, the pH value of solution is adjusted to 2.0, then adding hydrated barta to be neutralized to the pH value is 7.0;
(8) press filtration, decolouring: the neutralizer press filtration is removed to precipitation, and wash precipitation with clear water, add activated carbon decolorizing 1h in clear liquid, 60 ℃ of bleaching temperatures, the gac add-on is destainer 5%, then carries out the press filtration separation, removes filter residue, gets it and crosses cleaner liquid;
(9) filtrate purifying: adopt successively macropore acidulous acrylic acid cation exchange resin, macroreticular weakly base styrene series anion exchange resin, zwitterion mixed type exchange resin to process upper step solution;
(10) vacuum concentration, spraying drying, can obtain described silk gum aminoacids complex.
The molecular weight of the silk gum aminoacids complex that utilizes aforesaid method to obtain is 120 ~ 130D, weight loss on drying≤4%, ash content≤3%, arsenic content≤0.5mg/Kg, lead content≤4mg/Kg, mercury content≤0.08mg/Kg, total number of bacterial colony≤50cfu/g, intestinal bacteria≤15MPN/100g.Therefore the amino acid whose indices of silk gum, all higher than the internal trade industry standard SB/T10407-2007 of the People's Republic of China (PRC), is particularly suitable for the additive of food, medicine and makeup.
Claims (4)
1. the preparation method of a silk gum aminoacids complex, is characterized in that, comprises the steps:
(1) take husks and/or long telling as raw material, come unstuck after cleaning;
(2) reclaim come unglued liquid, husks and/or length are told to dehydration, then use deionized water wash, dehydration liquid, washings and come unglued liquid are merged, obtain silk gum solution, add 0.4 ~ 0.6% sanitas in silk gum solution;
(3) enzymolysis: first the pH value of upper step silk gum solution is adjusted into to 9 ~ 11, then use the above-mentioned silk gum solution of 2709 hydrolysis by novo, temperature is 40 ~ 50 ℃, enzyme concn is 750 ~ 800 unit of activity/g, reaction times 4 ~ 10h is total free aminoacids and part of polypeptide until detected result is major part;
(4) acidolysis: add 10 ~ 15% sulphuric acid hydrolysis in above-mentioned silk gum solution, micro-boiling reflux, be hydrolyzed 10 ~ 15h, until detected result is the overwhelming majority, is total free aminoacids and contains a small amount of dipeptides, tripeptides;
(5) neutralization: add CaO in the silk gum acid hydrolysis liquid obtained in upper step, the pH value of solution is adjusted to 1.5 ~ 2.5, then adding hydrated barta to be neutralized to the pH value is 7.0;
(6) press filtration, decolouring;
(7) adopt successively macropore acidulous acrylic acid cation exchange resin, macroreticular weakly base styrene series anion exchange resin, zwitterion mixed type exchange resin to process upper step solution;
(8) vacuum concentration, spraying drying, can obtain described silk gum aminoacids complex.
2. the preparation method of silk gum aminoacids complex according to claim 1, it is characterized in that: in described step (1), the bath raio of coming unstuck is 1:15 ~ 20, and pressure is 0.15 ~ 0.3MPa, and 100 ~ 140min comes unstuck under 120 ~ 140 ℃.
3. the preparation method of silk gum aminoacids complex according to claim 1, is characterized in that: in described step (3), first with sodium carbonate, the pH value of upper step silk gum solution is adjusted into to 9 ~ 11.
4. the silk gum aminoacids complex that preparation method according to claim 1 obtains.
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| CN102911994A (en) * | 2012-11-07 | 2013-02-06 | 广州优锐生物科技有限公司 | Hair hydrolysate and preparation method thereof |
| CN107219370B (en) * | 2017-07-21 | 2018-08-24 | 浙江理工大学 | A kind of radially-arranged method of amino acid in analysis Bombyx silk albumen |
| CN109289938A (en) * | 2018-10-30 | 2019-02-01 | 扬州金珠树脂有限公司 | The separation of sulfuric acid and the method for covering sour reuse in a kind of ion exchange resin hydrolyzate |
| CN109777846A (en) * | 2018-12-24 | 2019-05-21 | 蔡罗章 | A kind of preparation method for extracting amino acid from clover |
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| CN1101402C (en) * | 1998-12-01 | 2003-02-12 | 浙江丝绸工学院 | Sericin polypeptide extracted from frison, husks and waste silk and its extracting method |
| CN1410437A (en) * | 2002-02-11 | 2003-04-16 | 李学军 | Method of producing silk peptide, silk amino acid by acid hydrolysis of natural silk |
| KR100494229B1 (en) * | 2003-03-24 | 2005-06-13 | 김정호 | manufacturing process of sericin and sericin mask pack |
| 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 |
| CN101348821B (en) * | 2008-08-29 | 2011-08-31 | 湖州南方生物科技有限公司 | High purity sericin powder prepared from silkworm cocoon blaze and preparation thereof |
| JP5751745B2 (en) * | 2009-06-25 | 2015-07-22 | セーレン株式会社 | Sericin-derived polypeptide and stratum corneum peeling enzyme protective agent |
| CN102381919A (en) * | 2010-09-03 | 2012-03-21 | 香港纺织及成衣研发中心 | Grouping preparation method of amino acid groups with alkaline, acid and neutral functions |
| CN102206695A (en) * | 2011-04-02 | 2011-10-05 | 重庆海帆生化科技有限公司 | Method for preparing silk short peptides and silk amino acids by using waste silks |
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Address after: Suzhou City, Jiangsu province Xiangcheng District Ji Road 8 215137 Patentee after: Soochow University Address before: 215123 Suzhou City, Suzhou Province Industrial Park, No. love road, No. 199 Patentee before: Soochow University |
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