CN103755834B - A kind of method of preparing active peptide powder and chitin from shrimp crab accessory substance - Google Patents
A kind of method of preparing active peptide powder and chitin from shrimp crab accessory substance Download PDFInfo
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- CN103755834B CN103755834B CN201410036020.4A CN201410036020A CN103755834B CN 103755834 B CN103755834 B CN 103755834B CN 201410036020 A CN201410036020 A CN 201410036020A CN 103755834 B CN103755834 B CN 103755834B
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Abstract
Prepare a method for active peptide powder and chitin from shrimp crab accessory substance, belong to living resources High Efficient Machining Technology field. First by fresh shrimp and crab shells attrition crushing, then do not have shrimp and crab shells with water logging, add protease to carry out enzymolysis processing twice in ultrasonic wave after extraction process again, after enzymolysis finishes, drop goes out enzymolysis liquid for the second time; Go out the shrimp and crab shells of enzymolysis liquid for the second time at drop and wash to neutrality, then add acid to carry out decalcification processing, then obtain chitin after cleaning, being dried; The paste albumen of extruding during by attrition crushing and adipose tissue mix with enzymolysis liquid for the first time, after enzymolysis finishes, get solution for vacuum concentration, the more spray-dried active peptide powder that makes. The present invention can improve the utilization ratio of protease to greatest extent, improves the concentration of the protein peptides in enzymolysis liquid.
Description
Technical field
The invention belongs to living resources High Efficient Machining Technology field.
Background technology
Chitin is again chitin, chitin, is extensively present in the cartilage and fungi microbe cell membrane of the shell of the crustaceans such as shrimp and crab shells and the shell of various insects, part higher mammal, and be one of biomass resource the abundantest on the earth. Chitin is a kind of important natural macromolecule amylose polymer, and the product after deacetylation is called as shitosan. Chitin and chitosan has a wide range of applications in the fields such as food, chemical industry, medicine, agricultural.
At present prepare chitin with shrimp and crab shells and formed huge industry, but the preliminary working stage of chitin pollutes large, in a lot of places, ecological environment is produced serious influence, for this reason, carry out the energy-saving and emission-reduction of chitin, the technology upgrading of green manufacturing is the prerequisite of chitin, the sustainable development of shitosan industry.
The traditional handicraft of chitin is acid-base method, general high concentration (10～12%) hydrochloric acid solution that adopts is removed the mineral matter such as calcium, phosphorus in crustacean shell, water rinses to neutral again, then, with compositions such as the protein in sodium hydroxide solution hydrolysis shell and on shell, finally dehydrates and forms chitin. This soda acid treatment process, although method is simple, application is strong, produces wastewater flow rate very large, and particularly the protein in buck cannot effectively reclaim, and has serious environmental pollution. Conventionally produce one ton of chitin and will drop into 2.5-2.8 ton NaOH, 8-9 ton hydrochloric acid, approximately 350 tons of water, discharge is containing strong acid, highly basic and organic 250 tons of left and right of trade effluent. In the improvement of waste water for producing crust element, mainly adopt simple end treatment mode at present, economic input is large, and is difficult to reach pollution emission standard.
Thereby by the production technology of innovation chitin, start with from pretreatment of raw material link, could really realize cleaning of chitin production. Shrimp and crab shells adheres to a large amount of protein, fat, if by effectively reclaiming and use, can prepare and become the feed addictive that economic worth is larger, and can greatly reduce the input of follow-up soda acid, this is the important technology theory of cleaner production and the principle that must adhere to. Produce in recent years more biological recovery technique, substantially be to adopt enzyme process to carry out the hydrolysis of living beings, but effect is poor aspect application in practice, main cause is: the organic matter in (1) shrimp crab accessory substance have a type of combining closely with non-tight type, the biomass by hydrolyzation effect that biological enzyme is often combined closely to part is wherein bad, directly causes organic efficiency not high; (2) use cost of enzyme is high, and single enzyme preparation hydrolysis effect is bad, need to use complex enzyme. As for protease, its addition is generally 1% left and right of albumen in raw material, and common protease preparation, the market price is at 150-250 unit/kg, the cost of its input is obviously larger, if can not effectively reduce the consumption of enzyme preparation, is just difficult to bioanalysis is effectively applied; (3) subsequent treatment cost is high, is mainly that hydrolyzate generally needs concentrated processing, and the energy consumption of input is higher, and this is also that biological recovering method crust organic matter is difficult to one of bottleneck of breaking through.
Summary of the invention
The object of the invention is the problem existing for above-mentioned prior art, propose a kind of pollution-free, co-production of preparing cheaply active peptide powder and chitin from shrimp crab accessory substance.
The technology of the present invention comprises the following steps:
1) adopt roll-type extrusion equipment that the fresh shrimp and crab shells of removing sandstone and foreign material is carried out to attrition crushing, paste albumen and the adipose tissue obtaining respectively broken shrimp and crab shells and be extruded;
2) do not have broken shrimp and crab shells with water logging, first supersonic frequency higher than the ultrasonic wave of 20000Hz in extraction process 0.5～1h, and then add protease to carry out primary enzymolysis to process 2～4h; During enzymolysis processing, open off and on the ultrasonic wave of supersonic frequency higher than 20000Hz, each 5～10min; After enzymolysis finishes, drop goes out enzymolysis liquid for the first time;
3) in the broken shrimp and crab shells that drains enzymolysis liquid for the first time, add again the shrimp and crab shells of water to submergence fragmentation, again add protease to carry out secondary enzymolysis 0.5～1h, during enzymolysis processing, open off and on the ultrasonic wave of supersonic frequency higher than 20000Hz, each 5～10min; During enzymolysis, control temperature of reaction system≤50 DEG C; After enzymolysis finishes, drop goes out enzymolysis liquid for the second time;
4) chitin preparation: adding concentration in drop goes out the shrimp and crab shells of enzymolysis liquid is for the second time that NaOH or the sodium bicarbonate aqueous solution of 5～8wt% washs to neutrality, then adds acid to carry out decalcification processing, then obtain chitin through cleaning, after dry;
5) active peptide powder preparation: the paste albumen of extruding during by attrition crushing and adipose tissue mix with enzymolysis liquid for the first time, be to carry out enzymolysis processing 3～5h under the condition of 40～55 DEG C in mixed system temperature, after enzymolysis finishes, get solution for vacuum concentration, the more spray-dried active peptide powder that makes.
Superiority of the present invention is embodied in:
1, the ultrasonic wave producing by high-frequency generator can be propagated in solution, and make liquid flow and produce ten hundreds of micro-bubbles, its moment is broken the cavitation effect that produces with respect to thousands of atmospheric pressures, can promote shrimp and crab shells surface organic peel off and broken, promote enzymolysis process; Adopt ultrasonic wave-enzyme process assisted extraction, be conducive to accelerate protein from chitin superficial degradation, improve biological Deproteinated effect.
2, adopt secondary enzymolysis, strengthen biological Deproteinated effect, and by the reuse of secondary enzymolysis liquid, then when improving protease utilization rate, also further increased the concentration of the polypeptide in hydrolyzate.
3,, time prepared by chitin, because organic matter in the shrimp and crab shells through secondary enzymolysis liquid is removed in a large number, therefore the NaOH adding or sodium bicarbonate aqueous solution consumption are little, and alkali lye can repeatedly use.
4, utilize the first cutting out partial albumen of Mechanical Method organic matter, this part albumen finally carries out enzymolysis, makes full use of for the first time remaining prolease activity in enzymolysis liquid, makes protease focus on the de-albumen step of chitin, efficiently utilizes protease.
In a word, the present invention can improve the utilization ratio of protease to greatest extent, improves the concentration of the protein peptides in enzymolysis liquid.
In addition, the present invention, in the time of described attrition crushing, adds the water that accounts for shrimp and crab shells total amount 8～10%.
Described protease is bacillus subtilis or the letter Alcalase3.0T of Novi alkali protease.
Described acid is that concentration is the hydrochloric acid of 3～5wt%.
Detailed description of the invention
Raw material can adopt cray shell, or Macrobrachium rosenbergii shell, or the byproduct of the aquatic products processing such as other fresh shrimp, crab shell.
The protein active peptide of making can be used for feed addictive, and chitin is biomaterial.
Embodiment 1:
First the shrimp shell 100kg forming after fresh Macrobrachium rosenbergii processing is added to mechanical presses device, in extrusion process, add the water of 10kg left and right, obtain respectively albumen, the adipose tissue 25kg of paste, and broken shrimp shell.
Broken shrimp shell is put into 0.5m32.5kw ultrasonic extraction tank in, add water to make material submergence, first start ultrasonic device, in the ultrasonic wave that is 20010Hz in supersonic frequency, carry out ultrasonic extraction 1.0h. And then add protease 100g(bacillus subtilis, optimal pH 7.2, enzyme 50,000 units alive/g), controlled enzymatic hydrolysis temperature is carried out enzymolysis 2.0h for the first time under 45～50 DEG C of conditions, during the 2.0h of enzymolysis for the first time, supersonic frequency is the ultrasonic equipment intermittently starting of 20010Hz, each 10min, intermittently 10min. After enzymolysis finishes, drop goes out enzymolysis liquid 350L(S2 for the first time).
In in ultrasonic extraction tank, add water 350L again, and add protease 100g(bacillus subtilis, optimal pH 7.2, live 50,000 units/g) of enzyme carry out enzymolysis 1.0h for the second time, for the second time during enzymolysis, supersonic frequency is the ultrasonic equipment intermittently starting of 20010Hz, each 10min, intermittently 10min. During enzymolysis, temperature of reaction system is controlled at 45 DEG C for the second time. After end, drop goes out enzymolysis liquid 330L for the second time.
The batching water of sediment as next step will be removed after enzymolysis liquid filtration for the second time.
After twice enzymolysis processing of learning from else's experience, removing shrimp shell after living beings and adopt the sodium hydrate aqueous solution processing that mass percent is 5%, is under the condition of 90 DEG C, to be incubated 5.0h to wash to PH and be neutral in the temperature of mixed system. And then to add mass percent concentration be that 3.0% hydrochloric acid carries out decalcification, get solid content again through clear water washing, dry after, obtain chitin.
The albumen of the 25kg paste of extruding, adipose tissue are mixed with enzymolysis liquid for the first time, utilize protease residual in enzymolysis liquid for the first time to continue enzymolysis processing 3.0h, the temperature of controlled enzymatic hydrolysis system is 55 DEG C, after enzymolysis finishes, centrifugal removal insoluble matter, record solution solid content 3.2%, by Vacuum Concentration, to 80L, spraying is dried and obtains powdered protein active peptide 12kg.
Embodiment 2:
First fresh cray shell 100kg is added to mechanical presses device, extrude the living beings such as albumen, fat in shrimp shell. Extrusion process adds the water of 10kg left and right, obtains respectively albumen, the adipose tissue 30kg of paste, and broken shrimp shell.
Broken shrimp shell after extrusion process is put into 0.5M32.5kw ultrasonic extraction tank in, add the secondary hydrolyzate of water or last batch, make the complete submergence of broken shrimp shell, in the ultrasonic wave that is 20100Hz in supersonic frequency, carry out ultrasonic extraction 0.05h. Add again alkali protease 80g(Novi letter Alcalase3.0T), controlled enzymatic hydrolysis temperature, at 55 DEG C, is carried out enzymolysis 4.0h for the first time, for the first time during enzymolysis, ultrasonic equipment intermittently starting, each 5min, intermittently 5min. After enzymolysis finishes, drop goes out enzymolysis liquid 350L for the first time.
In in ultrasonic extraction tank, again add water 350L, secondary adds alkali protease (Novi letter Alcalase3.0T) 120g to enter enzymolysis 2.0h for the second time, for the second time during enzymolysis, and the ultrasonic equipment intermittently starting that supersonic frequency is 20100Hz, each 5min, intermittently 5min. During enzymolysis, 45 DEG C of temperature of reaction system. After end, drop goes out enzymolysis liquid 330L for the second time.
The batching water of sediment as next step will be removed after enzymolysis liquid filtration for the second time.
After twice enzymolysis processing of learning from else's experience, removing shrimp shell after living beings and adopt the sodium bicarbonate aqueous solution processing that mass percent is 5%, is to be incubated 6.0h under the condition of 90 DEG C in the temperature of mixed system, is neutral through washing to PH. And then to add mass percent concentration be that 5.0 hydrochloric acid carries out decalcification, get solid content again through clear water washing, dry after, obtain chitin.
The albumen of the 30kg paste of extruding, adipose tissue are mixed with enzymolysis liquid for the first time, utilize protease residual in enzymolysis liquid for the first time to continue enzymolysis 5.0h, 55 DEG C of temperature, after enzymolysis finishes, centrifugal removal insoluble matter, record solution solid content 2.5%, by Vacuum Concentration, to 60L, spraying is dried and obtains powdered protein active peptide 9.0kg.
Claims (1)
1. a method of preparing active peptide powder and chitin from shrimp crab accessory substance, is characterized in that comprising the following steps:
1) adopt roll-type extrusion equipment that the fresh shrimp and crab shells of removing sandstone and foreign material is carried out to attrition crushing, paste albumen and the adipose tissue obtaining respectively broken shrimp and crab shells and be extruded; In the time of described attrition crushing, add the water that accounts for shrimp and crab shells total amount 8～10%;
2) do not have broken shrimp and crab shells with water logging, first supersonic frequency higher than the ultrasonic wave of 20000Hz in extraction process 0.5～1h, and then add protease to carry out primary enzymolysis to process 2～4h; During enzymolysis processing, open off and on the ultrasonic wave of supersonic frequency higher than 20000Hz, each 5～10min; After enzymolysis finishes, drop goes out enzymolysis liquid for the first time;
3) in the broken shrimp and crab shells that drains enzymolysis liquid for the first time, add again the shrimp and crab shells of water to submergence fragmentation, again add protease to carry out secondary enzymolysis 0.5～1h, during enzymolysis processing, open off and on the ultrasonic wave of supersonic frequency higher than 20000Hz, each 5～10min; During enzymolysis, control temperature of reaction system≤50 DEG C; After enzymolysis finishes, drop goes out enzymolysis liquid for the second time;
4) chitin preparation: adding concentration in drop goes out the shrimp and crab shells of enzymolysis liquid is for the second time that NaOH or the sodium bicarbonate aqueous solution of 5～8wt% washs to neutrality, adding concentration is that the hydrochloric acid of 3～5wt% carries out decalcification processing again, then obtains chitin through cleaning, after dry;
5) active peptide powder preparation: the paste albumen of extruding during by attrition crushing and adipose tissue mix with enzymolysis liquid for the first time, be to carry out enzymolysis processing 3～5h under the condition of 40～55 DEG C in mixed system temperature, after enzymolysis finishes, get solution for vacuum concentration, the more spray-dried active peptide powder that makes;
Described protease is bacillus subtilis or the letter Alcalase3.0T of Novi alkali protease.
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| CN103990293B (en) * | 2014-05-28 | 2015-12-30 | 昆山市富众网络科技有限公司 | A kind of Carapax Eriocheir sinensis is efficiently broken, extraction equipment |
| CN104686779A (en) * | 2015-03-26 | 2015-06-10 | 岭南师范学院 | Low-cost production method for shrimp protein peptide |
| CN105249464B (en) * | 2015-11-18 | 2018-02-23 | 江南大学 | The one chelated calcium preparation method of seed shrimp peptide |
| CN109628525A (en) * | 2018-12-17 | 2019-04-16 | 上海冠硕生物科技有限公司 | A kind of technique that enzymatic hydrolysis chitin prepares N-acetylglucosamine |
| CN109893881A (en) * | 2019-01-24 | 2019-06-18 | 舟山瑞洋水产品研发有限公司 | Ocean shrimp crab extraction process and equipment |
| FR3105261A1 (en) * | 2019-12-19 | 2021-06-25 | Ynsect | Process for obtaining chitin and / or chitosan using two enzymatic hydrolyses |
| CN111657452A (en) * | 2020-06-09 | 2020-09-15 | 江苏省农业科学院 | Method for mechanically collecting crab meat and crab spawn |
| GB2612617A (en) * | 2021-11-05 | 2023-05-10 | Marine Bioenergy As | Demineralisation of organic tissue |
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