CN103627765B - A kind of preparation method of tea seed polypeptide - Google Patents
A kind of preparation method of tea seed polypeptide Download PDFInfo
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- CN103627765B CN103627765B CN201310631391.2A CN201310631391A CN103627765B CN 103627765 B CN103627765 B CN 103627765B CN 201310631391 A CN201310631391 A CN 201310631391A CN 103627765 B CN103627765 B CN 103627765B
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Abstract
The present invention relates to a kind of preparation method of tea seed polypeptide, specific as follows: tea seed albumen is through pulverizing, add water by mass volume ratio 3:100 ~ 120 after crossing 80 ~ 100 mesh sieves and make suspension, described suspension is in 90 ~ 95 DEG C of thermal treatment 10 ~ 20min, after cooling, adjust pH is to neutral, neutral protease is added by the enzyme concentration of 4300 ~ 4400u/g substrate, in 53 ~ 54 DEG C of enzymolysis 3.5 ~ 4.5h, go out after enzymolysis terminates enzyme, centrifugal supernatant liquor, described supernatant liquor is through desalination, after column chromatography, elutriant recentrifuge obtains supernatant liquor, described supernatant liquor obtains tea seed polypeptide through lyophilize.Extracting factor of the present invention is gentle, obtained camellia seed meal albumen form, color and luster are good, tea seed albumen is again through the enzymolysis of neutral protease, desalination and column chromatography, the elutriant of column chromatography obtains camellia seed meal polypeptide through lyophilize, this polypeptide impurities content is low, aminoacids content enriches, and can be widely used in foodstuff additive.
Description
Technical field
The present invention relates to the preparation method of a peptide species, relate to a kind of preparation method of tea seed polypeptide specifically.
Background technology
Oil tea belongs to evergreen dungarunga, is Theaceae Camellia, is a kind of excellent traditional oil tree of south China.China's oil tea resource is very abundant, and only the cultivated area of camellia oleifera lam just reaches 4,000,000 hm
2, be the country that camellia oleifera cultivar at most, distribution is the widest, tea seed output is the highest in the world, produce tea seed about 54.98 ten thousand tons per year, tea oil 150,000 tons, tea seed byproduct---the annual output of camellia seed meal is about 39.71 ten thousand tons.In camellia seed meal, protein content is about 12% ~ 15%, and its amino acid consists of 17 kinds, and wherein 8 kinds is essential amino acid.The utilization ratio of camellia seed meal is quite low, causes the significant wastage of resource, and plant protein is under biological enzyme condition, and the materials such as the polypeptide that hydrolysis generates and amino acid, have very high nutritive value and functional performance, therefore this method becomes study hotspot.The polypeptide produced after a variety of plant protein enzymolysis except there is easily digested characteristic, also have very strong anti-oxidant, reduce the functional performances such as cholesterol.Up to the present, have no both at home and abroad about the research of tea seed polypeptide is reported, therefore, carry out and the research of tea seed polypeptide nourishing function activity is had important practical significance.
Summary of the invention
The object of the invention is to solve the deficiencies in the prior art, a kind of preparation method of tea seed polypeptide is provided.
The technical solution adopted for the present invention to solve the technical problems is:
A preparation method for tea seed polypeptide, described preparation method is specific as follows:
Tea seed albumen is through pulverizing, add water by mass volume ratio 3:100 ~ 120 after crossing 80 ~ 100 mesh sieves and make suspension, described suspension is in 90 ~ 95 DEG C of thermal treatment 10 ~ 20min, after cooling, adjust pH is to neutral, neutral protease is added by the enzyme concentration of 4300 ~ 4400u/g substrate, in 53 ~ 54 DEG C of enzymolysis 3.5 ~ 4.5h, go out after enzymolysis terminates enzyme, centrifugal supernatant liquor, described supernatant liquor elutriant recentrifuge after desalination, column chromatography obtains supernatant liquor, and described supernatant liquor obtains tea seed polypeptide through lyophilize.
As preferably, the enzyme concentration of neutral protease is 4364u/g substrate, and hydrolysis temperature is 53.5 DEG C, and enzymolysis time is 4 hours.
As preferably, the preparation method of described tea seed albumen is as follows:
(1) enzymolysis: mix pulverizing the rear camellia seed meal powder crossing 80 ~ 100 mesh sieves with water by solid-liquid ratio 1:6 ~ 7, adjust pH to 7.25 ~ 7.30, add the composite plant lytic enzyme of camellia seed meal opaque amount 0.04 ~ 0.05%, in 65 ~ 70 DEG C of enzymolysis 3 ~ 3.5h, go out after enzymolysis terminates enzyme, then centrifugal acquisition camellia seed meal residue;
(2) extraction with ultrasonic-NaOH: the camellia seed meal residue that step (1) obtains is mixed with suitable quantity of water, adjust pH to 10.0 ~ 10.5,80 ~ 90min is carried in 40 ~ 45 DEG C of alkali, alkali is carried in process and is adopted ultrasonic assistant, hyperacoustic power is 300 ~ 1000W, ultrasonic time is 20min, and after alkali proposes end, centrifugation obtains supernatant liquor;
(3) acid is heavy: the supernatant liquor adjust pH to 4.6 step (2) obtained, and leaves standstill 1 ~ 1.5h in room temperature, then centrifugally must precipitate, and described precipitation obtains camellia seed meal albumen through lyophilize.
Nonprotein in camellia seed meal powder such as carbohydrate breakdown is first fallen by composite plant lytic enzyme by the present invention, and then carried by alkali, acid sinks to obtain camellia seed meal albumen, wherein alkali is carried and is passed through intensified by ultrasonic wave, extraction effect is good, the extraction yield that alkali is carried is high, because extracting factor of the present invention is gentle, obtained camellia seed meal albumen form, color and luster is good, tea seed albumen is again through the enzymolysis of neutral protease, desalination and column chromatography, the elutriant of column chromatography obtains camellia seed meal polypeptide through lyophilize, this polypeptide amino acid rich content, foodstuff additive can be widely used in.
As preferably, in step (1), the particle diameter of camellia seed meal powder is 80 orders, and solid-liquid ratio is 1:6, and pH value is 7.28, and the addition of composite plant lytic enzyme is 0.048%, and hydrolysis temperature is 68 DEG C, and enzymolysis time is 3.25h.The optimum process condition of this processing parameter system composite plant enzymatic hydrolysis, non-protein class removal of impurity is high, follow-up alkali carries the consumption can saving alkali in process, the consumption of alkali reduces, the destruction of camellia seed meal albumen is reduced, avoid alkalescence and cause too by force deamination, decarboxylation, peptide bond rupture, causing " Guang relies reaction ", is toxic compounds by amino acid.
As preferably, in step (2), camellia seed meal residue (in butt) is 1:25 with the mass ratio of water.
As preferably, the pH value that in step (2), alkali is carried is 10.0, and extraction temperature is 40 DEG C, and extraction time is 80min.
As preferably, described centrifugal rotating speed is 3000 ~ 3200rpm, and centrifugation time is 10 ~ 20min.
The invention has the beneficial effects as follows:
(1) such as the carbohydrate breakdown of the nonprotein in camellia seed meal powder is first fallen by composite plant lytic enzyme by the present invention, and then carried by alkali, acid sinks to obtain camellia seed meal albumen, wherein alkali is carried and is passed through intensified by ultrasonic wave, extraction effect is good, the extraction yield that alkali is carried is high, because extracting factor of the present invention is gentle, obtained camellia seed meal albumen form, color and luster is good, tea seed albumen is again through the enzymolysis of neutral protease, desalination and column chromatography, the elutriant of column chromatography obtains camellia seed meal polypeptide through lyophilize, this polypeptide impurities content is low, aminoacids content enriches, foodstuff additive can be widely used in.
(2) camellia seed meal that utilization ratio is quite low is made tea seed polypeptide by the present invention, has very high nutritive value, improves the utilization ratio of resource, is turned waste into wealth by camellia seed meal, adds its economic worth.
Embodiment
Below by specific embodiment, be described in further detail technical scheme of the present invention, the plant and instrument that each embodiment uses is all commercial conventional equipment, and the reagent that each embodiment uses is all commercial conventional reagent.
Composite plant lytic enzyme: Novozymes Company produces, enzyme 100FBG/g alive.
The aminoacids content of tea seed polypeptide prepared by the present invention is detected by test center of Inst. of Oil Crops, Chinese Academy of Agriculture.
Embodiment 1
A preparation method for tea seed polypeptide, described preparation method is specific as follows:
(1) enzymolysis: mix pulverizing the rear camellia seed meal powder crossing 80 mesh sieves with water by solid-liquid ratio 1:6, adjust pH to 7.25, add the composite plant lytic enzyme of camellia seed meal opaque amount 0.04%, in 65 DEG C of enzymolysis 3.5h, in 85 DEG C of enzyme 10min that go out after enzymolysis terminates, then obtain camellia seed meal residue in the centrifugal 15min of 3000rpm;
(2) extraction with ultrasonic-NaOH: the camellia seed meal residue (in butt) step (1) obtained in mass ratio 1:25 mixes with water, adjust pH to 10.0,90min is carried in 40 DEG C of alkali, alkali is carried in process and is adopted ultrasonic assistant, hyperacoustic power is 300W, ultrasonic time is 20min, is separated to obtain supernatant liquor after alkali proposes end in the centrifugal 15min of 3000rpm;
(3) acid is heavy: the supernatant liquor adjust pH to 4.6 step (2) obtained, leaves standstill 1h in room temperature, and then must precipitate in the centrifugal 15min of 3000rpm, described precipitation obtains camellia seed meal albumen through lyophilize.
(4) by mass volume ratio 3:100(g/ml after the tea seed albumen that step (3) obtains being crossed 80 mesh sieves) add water and make suspension, described suspension is in 90 DEG C of thermal treatment 20min, adjust pH to 7.0 after cooling, neutral protease is added by the enzyme concentration of 4300u/g substrate, in 53 DEG C of enzymolysis 4.5h, in 90 DEG C of enzyme 10min that go out after enzymolysis terminates, under 3000rpm speed conditions, centrifugal 20min obtains supernatant liquor again, described supernatant liquor is through desalination, after SephadexG-25 column chromatography elutriant again under 3000rpm speed conditions centrifugal 20min obtain supernatant liquor, described supernatant liquor obtains tea seed polypeptide through lyophilize.
Embodiment 2:
A preparation method for tea seed polypeptide, described preparation method is specific as follows:
(1) enzymolysis: mix pulverizing the rear camellia seed meal powder crossing 100 mesh sieves with water by solid-liquid ratio 1:7, adjust pH to 7.30, add the composite plant lytic enzyme of camellia seed meal opaque amount 0.05%, in 70 DEG C of enzymolysis 3h, in 85 DEG C of enzyme 15min that go out after enzymolysis terminates, then obtain camellia seed meal residue in the centrifugal 10min of 3200rpm;
(2) extraction with ultrasonic-NaOH: the camellia seed meal residue (in butt) step (1) obtained in mass ratio 1:25 mixes with water, adjust pH to 10.5,80min is carried in 45 DEG C of alkali, alkali is carried in process and is adopted ultrasonic assistant, hyperacoustic power is 1000W, ultrasonic time is 20min, is separated to obtain supernatant liquor after alkali proposes end in the centrifugal 10min of 3200rpm;
(3) acid is heavy: the supernatant liquor adjust pH to 4.6 step (2) obtained, leaves standstill 1.5h in room temperature, and then must precipitate in the centrifugal 10min of 3200rpm, described precipitation obtains camellia seed meal albumen through lyophilize.
(4) by mass volume ratio 3:120(g/ml after the tea seed albumen that step (3) obtains being crossed 100 mesh sieves) add water and make suspension, described suspension is in 95 DEG C of thermal treatment 10min, adjust pH to 7.0 after cooling, neutral protease is added by the enzyme concentration of 4400u/g substrate, in 54 DEG C of enzymolysis 3.5h, in 95 DEG C of enzyme 8min that go out after enzymolysis terminates, under 3200rpm speed conditions, centrifugal 15min obtains supernatant liquor again, described supernatant liquor is through desalination, after SephadexG-25 column chromatography elutriant again under 3200rpm speed conditions centrifugal 15min obtain supernatant liquor, described supernatant liquor obtains tea seed polypeptide through lyophilize.
Embodiment 3:
A preparation method for tea seed polypeptide, described preparation method is specific as follows:
(1) enzymolysis: mix pulverizing the rear camellia seed meal powder crossing 100 mesh sieves with water by solid-liquid ratio 1:6, adjust pH to 7.28, add the composite plant lytic enzyme of camellia seed meal opaque amount 0.048%, in 68 DEG C of enzymolysis 3.25h, in 90 DEG C of enzyme 5min that go out after enzymolysis terminates, then obtain camellia seed meal residue in the centrifugal 20min of 3000rpm;
(2) extraction with ultrasonic-NaOH: the camellia seed meal residue (in butt) step (1) obtained in mass ratio 1:25 mixes with water, adjust pH to 10.0,80min is carried in 40 DEG C of alkali, alkali is carried in process and is adopted ultrasonic assistant, hyperacoustic power is 800W, ultrasonic time is 20min, is separated to obtain supernatant liquor after alkali proposes end in the centrifugal 20min of 3000rpm;
(3) acid is heavy: the supernatant liquor adjust pH to 4.6 step (2) obtained, leaves standstill 1.5h in room temperature, and then must precipitate in the centrifugal 20min of 3000rpm, described precipitation obtains camellia seed meal albumen through lyophilize.
(4) by mass volume ratio 3:100(g/ml after the tea seed albumen that step (3) obtains being crossed 80 mesh sieves) add water and make suspension, described suspension is in 95 DEG C of thermal treatment 10min, adjust pH to 7.0 after cooling, neutral protease is added by the enzyme concentration of 4364u/g substrate, in 53.5 DEG C of enzymolysis 4h, in 95 DEG C of enzyme 8min that go out after enzymolysis terminates, under 3200rpm speed conditions, centrifugal 15min obtains supernatant liquor again, described supernatant liquor is through desalination, after SephadexG-25 column chromatography elutriant again under 3200rpm speed conditions centrifugal 15min obtain supernatant liquor, described supernatant liquor obtains tea seed polypeptide through lyophilize.
The organoleptics property index of the tea seed polypeptide that embodiment 1 ~ 3 obtains is in table 1, and the quality index of tea seed polypeptide is shown in Table 2:
The organoleptics property index of table 1 camellia seed meal polypeptide
The amino acid composition of the tea seed polypeptide that table 2 embodiment 1 ~ 3 is obtained and average content thereof
As shown in table 2, shown by table 2, the aminoacids content of tea seed polypeptide prepared by the present invention enriches, and is rich in multiple essential amino acid, is of high nutritive value.
The molecular weight distribution of the tea seed polypeptide that table 3 embodiment 1 ~ 3 is obtained
The molecular force of tea seed polypeptide detects and adopts HPLC method, shows that the molecular weight of tea seed polypeptide prepared by the present invention is mainly concentrated between 180Da ~ 600Da, account for 51.17 ~ 51.38% of whole tea seed polypeptide by table 3; Polypeptide between molecular weight 600Da ~ 1000Da accounts for 20.38 ~ 21.25% of whole tea seed polypeptide, what molecular weight was less than 180Da accounts for 19.23 ~ 20.01% of whole tea seed polypeptide, the molecular weight of tea seed polypeptide prepared by the present invention is little, absorption rate is high, being of high nutritive value of tea seed polypeptide.
Above-described embodiment is one of the present invention preferably scheme, not does any pro forma restriction to the present invention, also has other variant and remodeling under the prerequisite not exceeding the technical scheme described in claim.
Claims (6)
1. a preparation method for tea seed polypeptide, is characterized in that: described preparation method is specific as follows:
Tea seed albumen is through pulverizing, add water by mass volume ratio 3:100 ~ 120 after crossing 80 ~ 100 mesh sieves and make suspension, described suspension is in 90 ~ 95 DEG C of thermal treatment 10 ~ 20min, after cooling, adjust pH is to neutral, neutral protease is added by the enzyme concentration of 4300 ~ 4400u/g substrate, in 53 ~ 54 DEG C of enzymolysis 3.5 ~ 4.5h, go out after enzymolysis terminates enzyme, centrifugal supernatant liquor, described supernatant liquor elutriant recentrifuge after desalination, column chromatography obtains supernatant liquor, and described supernatant liquor obtains tea seed polypeptide through lyophilize; The preparation method of described tea seed albumen is as follows:
(1) enzymolysis: mix pulverizing the rear camellia seed meal powder crossing 80 ~ 100 mesh sieves with water by solid-liquid ratio 1:6 ~ 7, adjust pH to 7.25 ~ 7.30, add the composite plant lytic enzyme of camellia seed meal opaque amount 0.04 ~ 0.05%, in 65 ~ 70 DEG C of enzymolysis 3 ~ 3.5h, go out after enzymolysis terminates enzyme, then centrifugal acquisition camellia seed meal residue;
(2) extraction with ultrasonic-NaOH: the camellia seed meal residue that step (1) obtains is mixed with suitable quantity of water, adjust pH to 10.0 ~ 10.5,80 ~ 90min is carried in 40 ~ 45 DEG C of alkali, alkali is carried in process and is adopted ultrasonic assistant, hyperacoustic power is 300 ~ 1000W, ultrasonic time is 20min, and after alkali proposes end, centrifugation obtains supernatant liquor;
(3) acid is heavy: the supernatant liquor adjust pH to 4.6 step (2) obtained, and leaves standstill 1 ~ 1.5h in room temperature, then centrifugally must precipitate, and described precipitation obtains camellia seed meal albumen through lyophilize.
2. the preparation method of tea seed polypeptide according to claim 1, is characterized in that: the enzyme concentration of described neutral protease is 4364u/g substrate, and hydrolysis temperature is 53.5 DEG C, and enzymolysis time is 4 hours.
3. the preparation method of a kind of tea seed polypeptide according to claim 1 and 2, it is characterized in that: in step (1), the particle diameter of camellia seed meal powder is 80 orders, solid-liquid ratio is 1:6, pH value is 7.28, the addition of composite plant lytic enzyme is 0.048%, hydrolysis temperature is 68 DEG C, and enzymolysis time is 3.25h.
4. the preparation method of a kind of tea seed polypeptide according to claim 3, is characterized in that: in step (2), the mass ratio of camellia seed meal residue and water is 1:25, and the quality of camellia seed meal residue is in butt.
5. the preparation method of a kind of tea seed polypeptide according to claim 3, is characterized in that: the pH value that in step (2), alkali is carried is 10.0, and extraction temperature is 40 DEG C, and extraction time is 80min.
6. the preparation method of a kind of tea seed polypeptide according to claim 3, is characterized in that: described centrifugal rotating speed is 3000 ~ 3200rpm, and centrifugation time is 10 ~ 20min.
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| CN105166888A (en) * | 2015-08-17 | 2015-12-23 | 安徽省华银茶油有限公司 | Ginseng and pilose antler tea seed polypeptide lozenge and preparation method therefor |
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| CN111298185B (en) * | 2020-02-26 | 2022-03-15 | 黄山荷琇生物科技有限公司 | Camellia oil and tea seed meal antibacterial polypeptide medical dressing and preparation method and application thereof |
| CN111793666A (en) * | 2020-07-06 | 2020-10-20 | 安徽工程大学宣城产业技术研究院有限公司 | Tea seed polypeptide, enzymolysis preparation method thereof and antioxidant activity determination method |
| CN112772727A (en) * | 2021-01-25 | 2021-05-11 | 合肥工业大学 | Method for preparing tea seed protein beverage |
| CN114246247B (en) * | 2021-11-22 | 2024-10-01 | 煌上煌集团有限公司 | Preparation method of antioxidant oilseed tea seed peptide |
| CN114686552B (en) * | 2022-04-02 | 2024-04-30 | 吉林农业大学 | Preparation method of camellia seed meal polypeptide zinc chelate |
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Inventor after: He Dongping Inventor after: Liu Jingwei Inventor after: Hu Chuanrong Inventor after: Zhang Sihong Inventor after: Liu Lingyi Inventor after: Shuang Yang Inventor before: He Dongping Inventor before: Hu Chuanrong Inventor before: Shuang Yang Inventor before: Liu Jingwei |
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Effective date of registration: 20170527 Address after: 324000 Wujiang East Road, Shen Shen Economic Development Zone, Qujiang District, Zhejiang, Quzhou Co-patentee after: Wuhan Polytechnic University Patentee after: Quzhou Liu Jia Fragrant Food Co., Ltd. Address before: 324000 Wujiang East Road, Shen Shen Economic Development Zone, Qujiang District, Zhejiang, Quzhou Patentee before: Quzhou Liu Jia Fragrant Food Co., Ltd. |