CN101892280A - Preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides - Google Patents
Preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides Download PDFInfo
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- CN101892280A CN101892280A CN2010102145350A CN201010214535A CN101892280A CN 101892280 A CN101892280 A CN 101892280A CN 2010102145350 A CN2010102145350 A CN 2010102145350A CN 201010214535 A CN201010214535 A CN 201010214535A CN 101892280 A CN101892280 A CN 101892280A
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Abstract
The invention provides a preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides, which comprises the following steps: (1) selecting pinctada fucata martensii meat to be put into cold water with cold homogenate to obtain a pinctada fucata martensii meat homogenate solution; (2) adding alkali protease in the pinctada fucata martensii meat homogenate solution, mixing uniformly, and carrying out ultrasonic complex enzymolysis; (3) further conducting enzymolysis for the homogenate solution obtained by the ultrasonic complex enzymolysis to continue carrying out constant temperature enzymolysis in water bath, eliminating enzyme after finishing the enzymolysis, and obtaining supernate containing the pinctada fucata martensii meat anti-oxidizing oligopeptides by centrifugal separation after cooling; and (4) gradually separating the supernate containing the pinctada fucata martensii meat anti-oxidizing oligopeptides by adopting 5-10kDa and 1kDa ultrafiltration membranes through membrane separation and purification treatment and collecting the filter liquor to obtain the inctada fucata martensii meat anti-oxidizing oligopeptides solution, and carrying out purification treatment to obtain powder thereof. The method has simple operation and new technique, can greatly improve the hydrolysis rate of the pinctada fucata martensii meat by ultrasonic auxiliary enzymolysis, shortens hydrolysis time and reduces production cost.
Description
Technical field
The invention belongs to the sea-food processing technique field, be specifically related to a kind of preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides.
Background technology
Pinctada fucata belongs to the Pteriidae Pinctada, is the main shellfish kind of producing cultured pearl in the world.The pteria martensii aboundresources is economized in China Hainan, Guangdong, Guangxi etc.After middle nineteen sixties China pinctada fucata artificial breeding was achieved success and produced in enormous quantities, the pinctada fucata aquaculture was rapidly developed, cultured output from 1981 14.5 kilograms increase to 10000 kilograms of 2009.There are more than 1000 of sea water pearls plants in Hainan at present, and the pearl of producing accounts for 30～40% of national sea water pearls ultimate production.Pinctada fucata is opened and is produced a large amount of shellfish meat behind the pearl, and pinctada fucata meat rich in proteins, lipid content are low, and nutritive value is very high, has only at present to be processed as the Margarita jerky on a small quantity, and product form is single, and high-valued processing does not almost have.How rationally higher value application Carnis Margarita resource is a pinctada fucata aquaculture industry urgent problem.
Along with the development of modern society, the health that adds ambassador modern of the deterioration of living environment and social pressures is subjected to serious threat, and free radical causes the body injury sex change, causes the body various diseases to produce, and has caused the attention of people's intensive.The consumption of antioxidant is increasing; at a low price, efficiently, the exploitation of the antioxidant from natural food of low toxicity becomes the research focus; wherein with little, the easy absorption of molecular weight, active strong; can remove active oxygen radical superfluous in the body effectively; protection cell and mitochondrial normal configuration and function, the researchs such as generation that prevent lipid peroxidation are main.Natural antioxidants has a extensive future at medicine, makeup, healthcare products and aspects such as food and fodder additives, and screening has the new trend that the active natural resource of strong anti-oxidation have become researchs such as Food science, biology, medical science.At present less about the research of the preparation technology aspect of the anti-oxidant oligopeptides of pinctada fucata shellfish meat, also there is very big development space.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of pinctada fucata martensii meat anti-oxidizing oligopeptides, this method safety and environmental protection simple to operate, the shellfish meat that pinctada fucata is opened behind the pearl obtains higher value application, can improve the economic benefit of manufacturing enterprise.
Above-mentioned purpose of the present invention is achieved by the following technical solution: a kind of preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides contains following steps:
(1) cooled homogenate: choose and get shellfish meat homogenate after shellfish meat places pre-cold water homogenate;
(2) the ultrasonic wave complex enzyme hydrolysis is handled: add Sumizyme MP in shellfish meat homogenate, mixing, the mixed solution of above-mentioned shellfish meat homogenate and Sumizyme MP is placed ultrasonic instrument, regulating frequency is that 35kHz, power are 280～360W, and attemperation is 45～65 ℃ and carries out enzymolysis 50～70min;
(3) further enzymolysis: place 45～60 ℃ water-bath to continue constant temperature enzymolysis 0.5～2h the shellfish meat homogenate behind the ultrasonic wave complex enzyme hydrolysis, the enzymolysis supernatant liquor that enzyme is handled, cooling back centrifugation must contain pinctada fucata martensii meat anti-oxidizing oligopeptides that goes out that finishes;
(4) membrane sepn and purification process: it is that the ultra-filtration membrane of 5～10kDa and 1kDa progressively separates that the supernatant liquor that will contain pinctada fucata martensii meat anti-oxidizing oligopeptides adopts molecular weight cut-off successively, collect filtrate, promptly obtain pinctada fucata martensii meat anti-oxidizing oligopeptides liquid, purified handle the pinctada fucata martensii meat anti-oxidizing oligopeptides powder.
Among the above-mentioned preparation method:
Shellfish meat described in the step (1) needs through pre-treatment, and described preprocessing process is: choose freezing shellfish meat and thaw or scallop meat, remove internal organ, clean 2～3 times with clear water, more than the draining 5min.
Shellfish meat described in the step (1) is 1: 0.5～1.0 with the weightmeasurement ratio of pre-cold water.
The temperature of pre-cold water is 0～10 ℃ in the step (1), and the homogenate time is 2～5min.
The addition of the Sumizyme MP described in the step (2) is to add 1800～2500U in every gram shellfish meat homogenate.
The temperature of enzyme of going out in the step (3) is 100 ℃, and the time of the enzyme that goes out is 5～10min.
The process of centrifugation is in the step (3): the rotating speed of regulating refrigerated centrifuge is 10000～15000r/min, centrifugal 15～25min under 0～4 ℃.
The isolating condition of ultra-filtration membrane is in the step (4): working pressure is 3～4bar, and temperature is 3～10 ℃.
Preferably, the purification process process in the step (4) is: the pinctada fucata martensii meat anti-oxidizing oligopeptides liquid of gained is got the pinctada fucata martensii meat anti-oxidizing oligopeptides powder through lyophilize.
Preferably, the purification process process in the step (4) is: get the pinctada fucata martensii meat anti-oxidizing oligopeptides powder through concentrating under reduced pressure again after the pinctada fucata martensii meat anti-oxidizing oligopeptides liquid employing dehydrated alcohol dehydration with gained.
The present invention compared with prior art has following advantage:
(1) integrated technology that combines by ultrasonic wave complex enzyme hydrolysis and ultra-filtration technique of the present invention, pinctada fucata meat is carried out enzymolysis by Sumizyme MP, utilize ultrasonic technology to have the double attribute of fluctuation and energy, the one, cavitation phenomenon, the energy release that promptly in liquid medium, shows as the formation of bubble and break and follow, the 2nd, medium and container also can produce the second-order effect of resonance character by the absorption of sound, so promote that under hyperacoustic booster action Sumizyme MP is hydrolyzed to pinctada fucata meat, by the time of control enzymolysis, temperature, enzyme concentration, the material-water ratio example is obtained and is had the active oligopeptides of high anti-oxidation.A large amount of shellfish meat carries out higher value application after not only the main cultured pearl shellfish-pinctada fucata in the South Sea being opened pearl, and obtain having the anti-oxidant oligopeptides of functional health effect, can be widely used in light industry and medicines and health protections such as food, makeup, promote the Sustainable development of marine cultured pearl industry, have important economic implications and social effect.
(2) pinctada fucata meat carries out enzymolysis by Sumizyme MP, and degree of hydrolysis is about 33.8%, and the average peptide chain length is about 3.2, and the pinctada fucata martensii meat anti-oxidizing oligopeptides liquid that obtains is clear, colorless, and solid is a buff powder.Product has stronger anti-oxidant activity, and particularly the removing ability to DPPH free radical, superoxide anion, hydroxy radical qiao etc. is stronger, has good wettability power and moisture-retaining capacity.
(3) this technological operation is simple, and ultrasound-assisted enzymolysis can improve percent hydrolysis greatly, shortens hydrolysis time, and production cost is low, and product safety can improve the economic benefit of manufacturing enterprise.
Embodiment
The present invention will be described below to enumerate specific embodiment.It is pointed out that embodiment only is used for that the invention will be further described, do not represent protection scope of the present invention, nonessential modification and adjustment that other people prompting according to the present invention is made still belong to protection scope of the present invention.
Embodiment 1
The preparation method of the pinctada fucata martensii meat anti-oxidizing oligopeptides that present embodiment provides is as follows:
(1) shellfish meat pre-treatment: will freeze shellfish meat and thaw, and remove internal organ, and clean 2 times, and be placed in stainless steel or the frame of plastic more than the draining 5min with clear water;
(2) cooled homogenate: water is pre-chilled to 4 ℃ earlier, takes by weighing shellfish meat, volume ratio is 1: the 0.5 pre-cold water of adding by weight, places the homogenate 5min of tissue mashing machine, obtains shellfish meat homogenate;
(3) enzyme-added, ultrasonication: shellfish meat homogenate is placed stainless steel cup, add the letter food grade alkalescence protease A lcalase 2.4L of Novi proteolytic enzyme, Denmark NOVO company product, enzyme work is 2.0 * 10
4U/g adds enzyme by the weight of shellfish meat homogenate, adds the Alcalase 2.4L proteolytic enzyme of 2000U in every gram shellfish meat homogenate, after stirring evenly, be placed in the ultrasonic instrument, regulating the ultrasonic wave operating frequency is that 35kHz, hyperacoustic intensity are that 280W, control treatment temp are 45 ℃, and the time is 70min;
(4) further enzymolysis: after ultrasonication, the shellfish meat homogenate in the step (3) is continued at 45 ℃ of water bath with thermostatic control enzymolysis 2h the enzyme 10min that under 100 ℃, goes out, cooling rapidly behind the enzyme that goes out;
(5) centrifugal: the enzymolysis solution in the step (4) is centrifugal in refrigerated centrifuge, and the rotating speed of whizzer is 10000r/min, and 4 ℃ of following centrifugal 25min get supernatant liquor;
(6) ultrafiltration: utilize membrane separation technique that the supernatant liquor in the step (5) is separated, the working pressure of membrane sepn is 3.5bar, the ultrafiltration temperature is 4 ℃, is that the ultra-filtration membrane of 10kDa separates collection filtrate with molecular weight cut-off earlier, filtrate is that the ultra-filtration membrane of 5kDa further separates with molecular weight cut-off again, collect filtrate, filtrate is that the ultra-filtration membrane of 1kDa further separates collection filtrate with molecular weight cut-off again, promptly obtain pinctada fucata martensii meat anti-oxidizing oligopeptides liquid, be water white transparency;
(7) purifying: above-mentioned pinctada fucata martensii meat anti-oxidizing oligopeptides liquid is obtained white anti-oxidant oligopeptides powder through freeze-drying.
Embodiment 2
The preparation method of the pinctada fucata martensii meat anti-oxidizing oligopeptides that present embodiment provides is as follows:
(1) cooled homogenate: choose scallop meat, remove internal organ, clean 3 times with clear water, more than the draining 5min, the weightmeasurement ratio by the scallop meat of removing internal organ and pre-cold water is 1: 1.0 then, place behind 5 ℃ the pre-cold water homogenate 5min shellfish meat homogenate;
(2) the ultrasonic wave complex enzyme hydrolysis is handled: add Sumizyme MP in shellfish meat homogenate, Sumizyme MP can adopt commercially available bacillus licheniformis alkali protease, and its enzyme activity is 2.0 * 10
5～10 * 10
5U/g, other Sumizyme MP that can reach similar effect also can, the addition of bacillus licheniformis alkali protease is to add 2500U in every gram shellfish meat homogenate, mixing, the mixed solution of above-mentioned shellfish meat homogenate and bacillus licheniformis alkali protease is placed ultrasonic instrument, regulating frequency is that 35kHz, power are 300W, and attemperation is 50 ℃ and carries out enzymolysis 50min;
(3) further enzymolysis: place 50 ℃ water-bath to continue constant temperature enzymolysis 1.5h the shellfish meat homogenate behind the ultrasonic wave complex enzyme hydrolysis, the finish enzyme that goes out of enzymolysis is handled, the temperature of enzyme of going out is 100 ℃, go out time of enzyme is 8min, the cooling back is centrifugal, the rotating speed of whizzer is 12000r/min, and 2 ℃ of following centrifugal 20min get the supernatant liquor that contains pinctada fucata martensii meat anti-oxidizing oligopeptides;
(4) membrane sepn and purification process: the supernatant liquor that will contain pinctada fucata martensii meat anti-oxidizing oligopeptides carries out membrane sepn, the working pressure of membrane sepn is 4.0bar, the ultrafiltration temperature is 5 ℃, be that the ultra-filtration membrane of 8kDa separates earlier with molecular weight cut-off, collect filtrate, filtrate is that the ultra-filtration membrane of 5kDa further separates with molecular weight cut-off again, collect filtrate, filtrate is that the ultra-filtration membrane of 1kDa further separates with molecular weight cut-off again, collect filtrate, promptly obtain pinctada fucata martensii meat anti-oxidizing oligopeptides liquid, get the pinctada fucata martensii meat anti-oxidizing oligopeptides powder through concentrating under reduced pressure again after the pinctada fucata martensii meat anti-oxidizing oligopeptides liquid employing dehydrated alcohol dehydration with gained.
Embodiment 3
The preparation method of the pinctada fucata martensii meat anti-oxidizing oligopeptides that present embodiment provides is as follows:
(1) shellfish meat pre-treatment: will freeze shellfish meat and thaw, and remove internal organ, and clean 3 times, and be placed on draining 10min in the frame of plastic with clear water;
(2) cooled homogenate: water is pre-chilled to 8 ℃ earlier, takes by weighing shellfish meat, volume ratio is 1: the 0.8 pre-cold water of adding by weight, places the homogenate 4min of tissue mashing machine, obtains shellfish meat homogenate;
(3) enzyme-added, ultrasonication: shellfish meat homogenate is placed glass beaker, add the letter food grade alkalescence protease A lcalase 2.4L of Novi proteolytic enzyme, Denmark NOVO company product, enzyme work is 2.0 * 10
4U/g adds enzyme by the weight of shellfish meat homogenate, adds the Alcalase 2.4L proteolytic enzyme of 1800U in every gram shellfish meat homogenate, after stirring evenly, be placed in the ultrasonic instrument, regulating the ultrasonic wave operating frequency is that 35kHz, hyperacoustic intensity are that 360W, control treatment temp are 60 ℃, and the time is 60min;
(4) further enzymolysis: after ultrasonication, the shellfish meat homogenate in the step (3) is continued at 60 ℃ of water bath with thermostatic control enzymolysis 1.0h the enzyme 5min that under 100 ℃, goes out, cooling rapidly behind the enzyme that goes out;
(5) centrifugal: the enzymolysis solution in the step (4) is centrifugal in refrigerated centrifuge, and the rotating speed of whizzer is 15000r/min, and 0 ℃ of following centrifugal 15min gets supernatant liquor;
(6) ultrafiltration: utilize membrane separation technique that the supernatant liquor in the step (5) is separated, the working pressure of membrane sepn is 3bar, the ultrafiltration temperature is 10 ℃, be that the ultra-filtration membrane of 6kDa separates with molecular weight cut-off respectively earlier, collect filtrate, filtrate is that the ultra-filtration membrane of 1kDa further separates collection filtrate with molecular weight cut-off again, promptly obtain pinctada fucata martensii meat anti-oxidizing oligopeptides liquid, be water white transparency;
(7) purifying: above-mentioned pinctada fucata martensii meat anti-oxidizing oligopeptides liquid is obtained white anti-oxidant oligopeptides powder through freeze-drying.
Claims (10)
1. the preparation method of a pinctada fucata martensii meat anti-oxidizing oligopeptides is characterized in that: contain following steps:
(1) cooled homogenate: choose and get shellfish meat homogenate after shellfish meat places pre-cold water homogenate;
(2) the ultrasonic wave complex enzyme hydrolysis is handled: add Sumizyme MP in shellfish meat homogenate, mixing, the mixed solution of above-mentioned shellfish meat homogenate and Sumizyme MP is placed ultrasonic instrument, regulating frequency is that 35kHz, power are 280～360W, and attemperation is 45～65 ℃ and carries out enzymolysis 50～70min;
(3) further enzymolysis: place 45～60 ℃ water-bath to continue constant temperature enzymolysis 0.5～2h the shellfish meat homogenate behind the ultrasonic wave complex enzyme hydrolysis, the finish enzyme that goes out of enzymolysis is handled, and cooling back centrifugation must contain the supernatant liquor of pinctada fucata martensii meat anti-oxidizing oligopeptides;
(4) membrane sepn and purification process: it is that the ultra-filtration membrane of 5～10kDa and 1kDa progressively separates that the supernatant liquor that will contain pinctada fucata martensii meat anti-oxidizing oligopeptides adopts molecular weight cut-off successively, collect filtrate, promptly obtain pinctada fucata martensii meat anti-oxidizing oligopeptides liquid, purified handle the pinctada fucata martensii meat anti-oxidizing oligopeptides powder.
2. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1, it is characterized in that: the shellfish meat described in the step (1) needs through pre-treatment, described preprocessing process is: choose freezing shellfish meat and thaw or scallop meat, remove internal organ, clean 2～3 times with clear water, more than the draining 5min.
3. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1 is characterized in that: the shellfish meat described in the step (1) is 1: 0.5～1.0 with the weightmeasurement ratio of pre-cold water.
4. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1 is characterized in that: the temperature of pre-cold water is 0～10 ℃ in the step (1), and the homogenate time is 2～5min.
5. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1, it is characterized in that: the addition of the Sumizyme MP described in the step (2) is to add 1800～2500U in every gram shellfish meat homogenate.
6. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1, it is characterized in that: the temperature of the enzyme that goes out in the step (3) is 100 ℃, and the time of the enzyme that goes out is 5～10min.
7. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1 is characterized in that: the process of centrifugation is in the step (3): the rotating speed of regulating refrigerated centrifuge is 10000～15000r/min, at 0～4 ℃ of centrifugal 15～25min down.
8. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1 is characterized in that: the isolating condition of ultra-filtration membrane is in the step (4): working pressure is 3～4bar, and temperature is 3～10 ℃.
9. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1, it is characterized in that: the purification process process in the step (4) is: the pinctada fucata martensii meat anti-oxidizing oligopeptides liquid of gained is got the pinctada fucata martensii meat anti-oxidizing oligopeptides powder through lyophilize.
10. the preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides according to claim 1, it is characterized in that: the purification process process in the step (4) is: get the pinctada fucata martensii meat anti-oxidizing oligopeptides powder through concentrating under reduced pressure again after the pinctada fucata martensii meat anti-oxidizing oligopeptides liquid of gained is adopted the dehydrated alcohol dehydration.
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| CN102188332A (en) * | 2011-03-31 | 2011-09-21 | 中国科学院南海海洋研究所 | Cosmetic containing oceanic shellfish active peptide and preparation method and application thereof |
| CN102220402A (en) * | 2011-05-27 | 2011-10-19 | 中国水产科学研究院南海水产研究所 | Method for extracting antibacterial peptide from pearl oyster meat |
| CN102551112A (en) * | 2010-12-14 | 2012-07-11 | 大连工业大学 | Rapid preparation method of concentrate and powder of Antarctic krill on board |
| CN102559368A (en) * | 2010-12-14 | 2012-07-11 | 大连工业大学 | Preparation method of antarctic krill phospholipid |
| CN104673868A (en) * | 2015-03-18 | 2015-06-03 | 吉林大学 | Method for preparing wild clam meat protein source antioxidant peptide through ultrasound-assisted enzymolysis method |
| CN105821099A (en) * | 2016-04-15 | 2016-08-03 | 岭南师范学院 | Process for preparing antioxidant peptides from pinctada martensii by adopting fermentation method |
| CN109892661A (en) * | 2019-03-27 | 2019-06-18 | 集美大学 | A kind of pearl shell peptide and its production method |
| CN115057916A (en) * | 2022-07-06 | 2022-09-16 | 广州现代产业技术研究院 | Pinctada martensii meat antioxidant polypeptide and preparation method and application thereof |
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| CN102551112A (en) * | 2010-12-14 | 2012-07-11 | 大连工业大学 | Rapid preparation method of concentrate and powder of Antarctic krill on board |
| CN102559368A (en) * | 2010-12-14 | 2012-07-11 | 大连工业大学 | Preparation method of antarctic krill phospholipid |
| CN102551112B (en) * | 2010-12-14 | 2014-08-13 | 大连工业大学 | Rapid preparation method of concentrate and powder of Antarctic krill on board |
| CN102188332A (en) * | 2011-03-31 | 2011-09-21 | 中国科学院南海海洋研究所 | Cosmetic containing oceanic shellfish active peptide and preparation method and application thereof |
| CN102188332B (en) * | 2011-03-31 | 2013-04-24 | 中国科学院南海海洋研究所 | Cosmetic containing oceanic shellfish active peptide and preparation method and application thereof |
| CN102220402A (en) * | 2011-05-27 | 2011-10-19 | 中国水产科学研究院南海水产研究所 | Method for extracting antibacterial peptide from pearl oyster meat |
| CN104673868A (en) * | 2015-03-18 | 2015-06-03 | 吉林大学 | Method for preparing wild clam meat protein source antioxidant peptide through ultrasound-assisted enzymolysis method |
| CN105821099A (en) * | 2016-04-15 | 2016-08-03 | 岭南师范学院 | Process for preparing antioxidant peptides from pinctada martensii by adopting fermentation method |
| CN109892661A (en) * | 2019-03-27 | 2019-06-18 | 集美大学 | A kind of pearl shell peptide and its production method |
| CN115057916A (en) * | 2022-07-06 | 2022-09-16 | 广州现代产业技术研究院 | Pinctada martensii meat antioxidant polypeptide and preparation method and application thereof |
| CN115057916B (en) * | 2022-07-06 | 2024-05-07 | 广州现代产业技术研究院 | Pinctada martensii meat antioxidant polypeptide and preparation method and application thereof |
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