CN1335404A - Polypeptide preparing process - Google Patents
Polypeptide preparing process Download PDFInfo
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- CN1335404A CN1335404A CN 00109935 CN00109935A CN1335404A CN 1335404 A CN1335404 A CN 1335404A CN 00109935 CN00109935 CN 00109935 CN 00109935 A CN00109935 A CN 00109935A CN 1335404 A CN1335404 A CN 1335404A
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Abstract
The polypeptide preparing process uses alkali treated leather including the leftover as material and includes water washing; pre-treatment; mechanical crushing; enzymolysis with trypsin, microbe enzyme, neutral proteinase, lichen bacillus enzyme, pepsin and papain; heating to deactivate enzyme; filtering, desalting, vacuum concentration, spraying to dry, collection and packing. The process is environment friendly, produces no toxic pollutant, and has low cost, high yield, and high amino acid content in product.
Description
The invention belongs to field of biological product, specifically is the preparation technology of a peptide species.
Polypeptide (claiming polypeptide, collagen polypeptide again) contains 19 seed amino acids usually, and more than people lactoprotein's matter is amino acid contained two kinds, comprising adult essential seven seed amino acids and two kinds of required semi-dispensable amino acids of upgrowth and development of children.Polypeptide has effects such as high medical treatment, nutritive health-care, beauty and skin care.China's medicine, makeup etc. are required main by import.The import polypeptide is many to be raw material with fresh skin of mammal, adopts acid system or alkaline process to make, as Japanese Patent JP8027035A.Acid-hydrolysis method is rapid and thorough, but tryptophane is all destroyed, and Serine and tyrosine part are destroyed, and the product yield is low, and equipment corrosion is serious, and produces secondary pollution.The alkali hydrolysis method hydrolysis is also thorough rapidly, but the amino acid of hydroxyl and sulfydryl is all destroyed, and produces racemization (structure variation).Therefore the polypeptide produced of acid and alkali hydrolysis method because of the partial amino-acid structure variation, its biological activity, nutritive health-care, beauty functions are reduced.Chinese patent (CN1155549) and reported in literature, with the Mammals reticular tissue is raw material, adopt enzyme or organic acid to extract medical, used for cosmetic collagen protein and polypeptide, raw materials used costliness, cost height, technology is more loaded down with trivial details, the production cycle is long, the product yield is low.
The object of the present invention is to provide the preparation technology of a peptide species, adopt cheap raw material, the polypeptide of the no sex change of preparation is environmentally friendly under mild conditions, and the toxicological harmless pollutent produces, and production cost is low, product yield height.
The objective of the invention is to realize by following technical solution:
With the secondary refuse salkali waste skin corner of process hides or/and the alkali double-layer fur is a raw material, after washing, with physics and the chemical process pretreating raw material that combines, pretreatment technology is: 100 parts of raw materials, add 40～60 parts in water, 5～20 parts of ammonium chlorides, 1～4 part of hydrochloric acid, handled 30～90 minutes, water cleans up, and the raw material after the cleaning adds 100～200 parts in water, 0.5～2 part of Emulsifier O, 0.5～2 part of emulsifying agent Tween-65, normal temperature was handled 30～90 minutes, water cleans up, pretreated raw material is broken into fritter, adds 80～150 parts in water, 3～15 parts of NaCl, 0～5 part of hydrochloric acid, 66%H
2SO
40～1.5 part, handled 1～6 hour, remove liquid, water cleans, add 0.5～1 part of NaOH, transfer pH6.5～7.5 standby, make the materials with hide glue fibril bundle fully loose, some key is interrupted and is separated, then be converted into uniform tropocollagen dispersion system, carry out enzymolysis again, enzymolysis process is: get 100 parts of raw materials after the processing, add 50～150 parts in water, temperature is 35～40 ℃, 0.5～0.8 part in trypsinase reacted 30～90 minutes, added 0.8～1 part of microbial enzyme and neutral protease, a kind of in the Bacillus licheniformis enzyme or two kinds, wherein neutral protease is 0.5～0.7 part, 0.2～0.5 part of Bacillus licheniformis enzyme, adjust pH 8.5～9 is warmed up to 45～50 ℃, be incubated 30～90 minutes, add stomach en-, a kind of in the papoid or two kinds, wherein stomach en-is 0.1～0.4 part, 0.3～0.7 part of papoid, handled 2～4 hours, at 80～90 ℃, heated 10～20 minutes then, make enzyme deactivation, again after filtration, desalting purifying, vacuum concentration, spraying drying is collected under aseptic condition, packing promptly gets polypeptide.
Existing production is medical, edible, the used for cosmetic polypeptide is fresh animal or bone, and the raw material that present technique adopts is industrial secondary refuse alkali skin tailing and alkali double-layer fur that tanning production reclaims, turn harm into good, turn waste into wealth, thereby the former cost height, raw material is limited, and latter's cost is low, raw material is very abundant, and market potential is big.
One of gordian technique of the present invention be loose with physical method and relatively more weak acid-base pretreatment, separate the materials with hide glue fibril bundle, for endonuclease reaction efficiently, homogeneous established good basis.Because be the weak acid alkali pre-treatment of under normal temperature condition (10～30 ℃), will not cause constituting the variation of amino-acid residue recurring structure.
Two of gordian technique is that abstraction reaction is (35～50 ℃) at low temperatures, with have strictness optionally biological enzyme formulation on purpose extract, easy to control.The composition structure of product, performance stable effect, it is little to make a variation.And the acid-alkali treatment method under the hot conditions, the arbitrariness of processing is very big, and the molecular weight distribution of product is wide, form structure, performance effect difference is big, even the partial amino-acid residue is destructurized, loses its specific biological activity.
Do not see that both at home and abroad the secondary refuse of useful rawhide prepares the patent documentation report of polypeptide.
The present technique remarkable in economical benefits, social environment benefit is outstanding, China's year leather processing amount accounts for 1/5 of the world, rank first in the world, produce not about 700,000 tons of the secondary refuse of tanning rawhide every year,, not only cause billions of first financial losses because of suitably being utilized, and rot and cause great environmental pollution, seriously have influence on the Sustainable development of tanning industry.Present technique is converted into the polypeptide of high added value with the secondary refuse of process hides, and its total amino acid content surpasses 90%.Not only can create remarkable economic efficiency, and not produce secondary pollution substantially in the production process, belong to environmental friendliness and green chemical industry industry, this plays a positive role to the benign development that promotes industry such as husbandry, process hides and leather goods.
Be embodiments of the invention below.
Embodiment one
Choose the salkali waste skin bit (corner) that produces in the leather processing procedure, with or tap water clean up.
100g alkali skin bit adds water 40g, ammonium chloride 5g, hydrochloric acid 2g processing 60 minutes, cleans up with deionized water.
Raw material after the cleaning adds deionized water 120g, Emulsifier O 0.5g, emulsifying agent Tween-65 1g, and normal temperature was handled 80 minutes; Clean up with deionized water.
Be cut into small pieces with machinery.
Add deionized water 100g, NaCl 10g, HCl 2g, 66%H
2SO
40.8g, handled 3 hours, remove liquid, washed with de-ionized water.
Add NaOH0.5g, transfer pH6.5～7.5 standby.
Get the raw material 100g after the processing, add deionized water 100g, be warmed to 40 ℃, trypsinase 0.6g reacted 30 minutes, added AS1.398 microbial enzyme 1g, 3492 neutral protease 0.5g, 2709 Bacillus licheniformis enzyme 0.3g, adjust pH 8.5～9, be warmed up to 45 ℃, be incubated 60 minutes, use the trichoroacetic acid(TCA) inspection, add stomach en-0.1g, papoid 0.5g handled 2 hours.
Be warmed up to 85 ℃, heated 10 minutes, cool to 40 ℃, filtered while hot.
Filtrate is crossed dialysis tubing.
Vacuum concentration (vacuum tightness 0.06～0.08,70～80 ℃ of temperature) is 30～35% to content of peptides, promptly gets and concentrates polypeptide products, and recording 19 seed amino acid total contents is 91%.
To concentrate the polypeptide liquid spraying drying, promptly get the pulverulent solids polypeptide products.
Embodiment two
Choose the alkali double-layer fur that produces in the leather processing procedure, clean up with deionized water.
100g alkali double-layer fur adds water 50g, ammonium chloride 10g, hydrochloric acid 1g processing 90 minutes, cleans up with deionized water.
Raw material after the cleaning adds deionized water 200g, Emulsifier O 1g, emulsifying agent Tween-65 2g, and normal temperature was handled 30 minutes; Clean up with deionized water.
Be cut into small pieces with machinery.
Add deionized water 80g, NaCl 3g, 66%H
2SO
41g handled 4 hours, removed liquid, washed with de-ionized water.
Add NaOH0.8g, transfer pH6.5～7.5 standby.
Get the raw material 100g after the processing, add deionized water 50g, be warmed to 35 ℃, add trypsinase 0.5g, reacted 60 minutes, and added AS1.398 microbial enzyme 1g, 3492 neutral protease 0.7g, adjust pH 8.5～9, be warmed up to 50 ℃, be incubated 30 minutes, check with trichoroacetic acid(TCA), add stomach en-0.4g, handled 4 hours.
Be warmed up to 80 ℃, heated 15 minutes, cool to 40 ℃, filtered while hot.
Filtrate is crossed dialysis tubing.
Vacuum concentration (vacuum tightness 0.06～0.08,70～80 ℃ of temperature) is 30～35% to content of peptides, promptly gets and concentrates polypeptide products.
To concentrate the polypeptide liquid spraying drying, and promptly get the pulverulent solids polypeptide products, recording 19 seed amino acid total contents is 90%.
Embodiment three
Choose the salkali waste skin bit (corner) that produces in the leather processing procedure, clean up with deionized water.
100g alkali skin bit adds water 60g, ammonium chloride 20g, hydrochloric acid 3g, handles 30 minutes, cleans up with deionized water.
Raw material after the cleaning adds deionized water 100g, Emulsifier O 1g, emulsifying agent Tween-65 1.0g, and normal temperature was handled 90 minutes; Clean up with deionized water.
Be cut into small pieces with machinery.
Add deionized water 150g, NaCl 15g, HCl 5g, 66%H
2SO
41.5g, handled 1 hour, remove liquid, washed with de-ionized water.
Add NaOH1g, transfer pH6.5～7.5 standby.
Get the raw material 100g after the processing, add deionized water 150g, be warmed to 40 ℃, trypsinase 0.8g reacted 60 minutes, added AS1.398 microbial enzyme 0.8g, 2709 Bacillus licheniformis enzyme 0.5g, adjust pH 8.5～9 is warmed up to 45 ℃, is incubated 90 minutes, check with trichoroacetic acid(TCA), add stomach en-0.3g, papoid 0.5g handled 3 hours.
Be warmed up to 90 ℃, heated 10 minutes, cool to 40 ℃, filtered while hot.
Filtrate is crossed dialysis tubing.
Vacuum concentration (vacuum tightness 0.06～0.08,70～80 ℃ of temperature) is 30～35% to content of peptides, promptly gets and concentrates polypeptide products.
To concentrate the polypeptide liquid spraying drying, and promptly get the pulverulent solids polypeptide products, recording 19 seed amino acid total contents is 91%.
Embodiment four
Choose the salkali waste skin bit that produces in the leather processing procedure, clean up with tap water.
100g alkali skin bit adds water 50g, ammonium chloride 10g, hydrochloric acid 4g processing 30 minutes, cleans up with deionized water.
Raw material after the cleaning adds deionized water 120g, Emulsifier O 2g, emulsifying agent Tween-65 0.5g, and normal temperature was handled 80 minutes; Clean up with deionized water.
Be cut into small pieces with machinery.
Add deionized water 150g, NaCl 15g, HCl 0.5g handled 6 hours, removed liquid, washed with de-ionized water.
Add NaOH1g, transfer pH6.5～7.5 standby.
Get the raw material 100g after the processing, add deionized water 100g, be warmed to 35 ℃, trypsinase 0.5g reacted 90 minutes, added AS1.398 microbial enzyme 0.9g, 3492 neutral protease 0.5g, 2709 Bacillus licheniformis enzyme 0.2g, adjust pH 8.5～9 is warmed up to 45 ℃, be incubated 60 minutes, use the trichoroacetic acid(TCA) inspection, papoid 0.7g handled 2 hours.
Be warmed up to 80 ℃, heated 20 minutes, cool to 40 ℃, filtered while hot.
Filtrate is crossed dialysis tubing.
Vacuum concentration (vacuum tightness 0.06～0.08,70～80 ℃ of temperature) is 30～35% to content of peptides, promptly gets and concentrates polypeptide products.
To concentrate the polypeptide liquid spraying drying, and promptly get the pulverulent solids polypeptide products, recording 19 seed amino acid total contents is 90%.
Embodiment five
Choose the salkali waste skin bit (corner) that produces in the leather processing procedure, clean up with tap water.
100g alkali skin bit adds water 60g, ammonium chloride 8g, hydrochloric acid 2g processing 60 minutes, cleans up with deionized water.
Raw material after the cleaning adds deionized water 120g, Emulsifier O 0.8g, emulsifying agent Tween-65 0.5g, and normal temperature was handled 50 minutes; Clean up with deionized water.
Be cut into small pieces with machinery.
Add deionized water 130g, NaCl 10g, HCl 2g, 66%H
2SO
40.5g, handled 3 hours, remove liquid, washed with de-ionized water.
Add NaOH0.5g, transfer pH6.5～7.5 standby.
Get the raw material 100g after the processing, add deionized water 100g, be warmed to 35 ℃, trypsinase 0.6g reacted 60 minutes, added AS1.398 microbial enzyme 0.8g, 3492 neutral protease 0.5g, 2709 Bacillus licheniformis enzyme 0.2g, adjust pH 8.5～9, be warmed up to 45 ℃, be incubated 60 minutes, use the trichoroacetic acid(TCA) inspection, add stomach en-0.3g, papoid 0.3g handled 4 hours.
Be warmed up to 85 ℃, heated 15 minutes, cool to 40 ℃, filtered while hot.
Filtrate is crossed dialysis tubing.
Vacuum concentration (vacuum tightness 0.06～0.08,70～80 ℃ of temperature) is 30～35% to content of peptides, promptly gets and concentrates polypeptide products.
To concentrate the polypeptide liquid spraying drying, and promptly get the pulverulent solids polypeptide products, recording 19 seed amino acid total contents is 91%.
Claims (1)
1. the preparation technology of polypeptide, it is characterized in that with secondary refuse in alkali skin corner and/or alkali double-layer fur be raw material, after washing, carry out pre-treatment, pretreatment technology is: 100 parts of raw materials, add 40～60 parts in water, 5～20 parts of ammonium chlorides, 1～4 part of hydrochloric acid, handled 30～90 minutes, water cleans up, and the raw material after the cleaning adds 100～200 parts in water, 0.5～2 part of Emulsifier O, 0.5～2 part of emulsifying agent Tween-65, normal temperature was handled 30～90 minutes, and water cleans up; Pretreated raw material is broken into fritter, adds 80～150 parts in water, 3～15 parts of NaCl, 0～5 part of hydrochloric acid, 66%H
2SO
40～1.5 part, handled 1～6 hour, remove liquid, water cleans, and adds 0.5～1 part of NaOH, transfers pH6.5～7.5 standby; Carry out enzymolysis again, enzymolysis process is: get 100 parts of raw materials after the processing, add 50～150 parts in water, temperature is 35～40 ℃, 0.5～0.8 part in trypsinase reacted 30～90 minutes, added 0.8～1 part of microbial enzyme and neutral protease, a kind of in the Bacillus licheniformis enzyme or two kinds, wherein neutral protease is 0.5～0.7 part, 0.2～0.5 part of Bacillus licheniformis enzyme, adjust pH 8.5～9 is warmed up to 45～50 ℃, be incubated 30～90 minutes, add stomach en-, a kind of in the papoid or two kinds, wherein stomach en-is 0.1～0.4 part, 0.3～0.7 part of papoid, handled 2～4 hours, at 80～90 ℃, heated 10～20 minutes then, make enzyme deactivation, again after filtration, desalting purifying, vacuum concentration, spraying drying is collected under aseptic condition, packing promptly gets polypeptide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00109935 CN1335404A (en) | 2000-07-21 | 2000-07-21 | Polypeptide preparing process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00109935 CN1335404A (en) | 2000-07-21 | 2000-07-21 | Polypeptide preparing process |
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| CN1335404A true CN1335404A (en) | 2002-02-13 |
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| CN 00109935 Pending CN1335404A (en) | 2000-07-21 | 2000-07-21 | Polypeptide preparing process |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101805772A (en) * | 2010-03-26 | 2010-08-18 | 东华大学 | Method for extracting protein from excess sludge |
| CN102153378A (en) * | 2010-11-24 | 2011-08-17 | 北京化工大学 | Method for preparing amino acid and small molecular peptide composite organic fertilizer and recycling chromium from leather waste |
| CN101613735B (en) * | 2009-08-17 | 2011-08-24 | 张吉越 | Method for preparing organic polypeptide |
-
2000
- 2000-07-21 CN CN 00109935 patent/CN1335404A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101613735B (en) * | 2009-08-17 | 2011-08-24 | 张吉越 | Method for preparing organic polypeptide |
| CN101805772A (en) * | 2010-03-26 | 2010-08-18 | 东华大学 | Method for extracting protein from excess sludge |
| CN102153378A (en) * | 2010-11-24 | 2011-08-17 | 北京化工大学 | Method for preparing amino acid and small molecular peptide composite organic fertilizer and recycling chromium from leather waste |
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Effective date of registration: 20030804 Address after: 610065 Sichuan province Chengdu City Ring Road No. 24 Sichuan University West South Garden 8 Building 3 unit 43 Applicant after: Zhang Qiang Address before: 610065 Sichuan province Chengdu City Ring Road No. 24 Sichuan University West South Garden 8 Building 3 Unit No. 43 Zhang Qiang Applicant before: Zhang Mingrang |
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