CN104215590A - Method for quickly detecting oligopeptide content - Google Patents
Method for quickly detecting oligopeptide content Download PDFInfo
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- CN104215590A CN104215590A CN201410410849.6A CN201410410849A CN104215590A CN 104215590 A CN104215590 A CN 104215590A CN 201410410849 A CN201410410849 A CN 201410410849A CN 104215590 A CN104215590 A CN 104215590A
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- oligopeptide
- enzymolysis liquid
- trichloroacetic acid
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- transmittance
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Abstract
The invention relates to the field of oligopeptide technology processing, and in particular relates to a method for quickly detecting the oligopeptide content. The method is characterized in that the hydrolysis degree of peptides and the oligopeptide content in an oligopeptide production process are quickly detected according to a difference of sedimentation effects of trichloroacetic acid on polypeptide chains with different molecular weights, so that oligopeptide can be effectively obtained. The method disclosed by the invention is quick and simple; the detection process is easy to control; the detection time is greatly shortened; the environment pollution is reduced, and the detection cost is lowered. The method can be used for oligopeptide product production control and is suitable for center control on industrial production.
Description
Technical field
The present invention relates to oligopeptide processing technique field, particularly a kind of in oligopeptide enzymolysis production run, carry out the method for oligopeptide content fast detecting, controlled hydrolysis degree.
Background technology
Amino acid is the basic comprising unit of protein.Different types of amino acid, according to specific order, forms the protein of structure difference, Various Functions.It is generally acknowledged that protein is the Amino acid profile above by 51.And in life entity, also having the material by 2～50 Amino acid profiles, scientific circles are referred to as " peptide ".Peptide is the material between amino acid and protein.Amino acid whose molecule minimum, protein maximum, thus two or more amino acid dehydrating condensations forms a peptide of several peptide bond compositions, and multiple peptides carry out the multistage folding protein molecule that just forms.Conventionally, by the oligopeptide that is called by 2～10 Amino acid profiles, by the polypeptide that is called being formed by 11～50 amino acid.On material forms, peptide and protein have homogeney, but there is unique physiological function, the especially oligopeptide of different aminoacids and protein, the characteristics such as it has, and molecular weight is little, good water solubility, Heat stability is good, good, the direct absorbability of absolute acid stability, high safety.Can be applied to there is the food of nutrition curative effect and low irritability food, health food, sports food, high-protein food, bakery product, candy, cake, cold drink etc.
Oligopeptide is produced at present, mainly adopts proteolytic enzyme protolysate mode, and directional cutting long-chain protein or polypeptide are produced until form the oligopeptide mode of 2～10 Amino acid profiles.But protease hydrolytic degree is wayward, hydrolysis degree is crossed the shallow remaining a large amount of large molecular polypeptides in enzymolysis liquid that cause, and crosses and deeply can produce again a large amount of free amino acids as hydrolysis degree.Need the modes such as following adopted ultrafiltration, nanofiltration to enzymolysis liquid processing, obtain the oligopeptide of specified molecular weight, the production cycle is long, and affects product yield and quality.And use the modes such as conventional gel chromatography to carry out molecular weight detection length consuming time, cannot synchronously monitor hydrolysis degree, on-line monitoring.A kind of content of can method in real time, fast and accurately measuring oligopeptide in production run of visible urgent need.
Summary of the invention
The object of the present invention is to provide a kind of method of oligopeptide content fast detecting, synchro control protein liquid hydrolysis degree, in-service monitoring hydrolysis result.Effectively produce oligopeptide product, without carrying out subsequent ultrafiltration, nanofiltration operation, shortened technological process, improved product yield, solved existing detection method cycle length, affected the problems such as product quality.
For achieving the above object, the technical solution used in the present invention is: a kind of method for quick of oligopeptide content, and its step comprises:
One, enzymolysis liquid pre-service: after enzymolysis liquid filters, add after trichloroacetic acid precipitation centrifugally, obtain clarified supernatant;
Two, transmittance detects: centrifuged supernatant detects transmittance with spectrophotometer.
Preferably, the method for a kind of oligopeptide content fast detecting described in step 1, is characterized in that the described enzymolysis liquid of step (1) is the enzymolysis liquid being hydrolyzed, do not go out enzyme, and enzymolysis liquid uses 3M Medium speed filter paper to filter to obtain clear filtrate.In process, use solid content detector to detect solid content in enzymolysis liquid, regulator solution solid content is within the scope of 5-15%.Trichloroacetic acid concentration 30% used, to trichloroacetic acid concentration in solution be 10-20%, mix, under treating fluid normal temperature condition, the centrifugal 10min of 4000rpm, obtains clarified supernatant.
Preferably, the method for a kind of oligopeptide content fast detecting described in step 2, is characterized in that it is 500nm that the described transmittance of step (2) detects wavelength, uses the clarification filtration liquid that does not add trichloroacetic acid processing that step 1 obtains as reference.As at the bottom of carrying out the sample hose after centrifugal or on wall without naked eyes visible precipitate, and use spectrophotometer to detect transmittance to reach more than 85%, can judge that the oligopeptide content in former enzymolysis liquid clear liquid exceedes 90% of Tot Prot.
The invention has the beneficial effects as follows: use the inventive method to detect the oligopeptide content in enzymolysis liquid, have fast, advantage simply and accurately, can be used for the control of production intermediate link, effectively shorten sense cycle, enhance productivity.
Embodiment
Embodiment 1
Use 3M Medium speed filter paper to filter to obtain clear filtrate the enzymolysis liquid being hydrolyzed.Use solid content detector to detect the solid content in enzymolysis liquid, regulator solution solid content is 5%.In clarification filtration liquid, add 30% trichloroacetic acid solution to trichloroacetic acid final concentration 10%, mix, pack in transparent conical centrifuge tube, centrifugal 10min under normal temperature, revolution 4000 rpm conditions; Supernatant carries out transmittance detection under 500 nm conditions.
Embodiment 2
Use 3M Medium speed filter paper to filter to obtain clear filtrate the enzymolysis liquid being hydrolyzed.Use solid content detector to detect the solid content in enzymolysis liquid, regulator solution solid content is 10%.In clarification filtration liquid, add 30% trichloroacetic acid solution to trichloroacetic acid final concentration 15%, mix, pack in transparent conical centrifuge tube, centrifugal 10min under normal temperature, revolution 4000 rpm conditions, supernatant carries out transmittance detection under 500 nm conditions.
Embodiment 3
Use 3M Medium speed filter paper to filter to obtain clear filtrate the enzymolysis liquid being hydrolyzed.Use solid content detector to detect the solid content in enzymolysis liquid, regulator solution solid content is 15%.In clarification filtration liquid, add 30% trichloroacetic acid solution to trichloroacetic acid final concentration 20%, mix, pack in transparent conical centrifuge tube, centrifugal 10min under normal temperature, revolution 4000 rpm conditions; Supernatant carries out transmittance detection under 500 nm conditions.
The above, be only the specific embodiment of the present invention, is not limited to this, any be familiar with those skilled in the art the present invention disclose technical scope in, can expect easily change or replace, within all should being encompassed in protection scope of the present invention.
Claims (3)
1. a method for oligopeptide content fast detecting, comprises the following steps:
(1) enzymolysis liquid pre-service, enzymolysis liquid adds trichloroacetic acid precipitation centrifugal after filtering;
(2) transmittance detects, and centrifuged supernatant detects transmittance with spectrophotometer.
2. the method for a kind of oligopeptide content fast detecting according to claim 1, it is characterized in that the described enzymolysis liquid of step (1) is the enzymolysis liquid being hydrolyzed, do not go out enzyme, enzymolysis liquid uses 3M Medium speed filter paper to filter to obtain clear filtrate, in process, use solid content detector to detect solid content in enzymolysis liquid, regulator solution solid content is within the scope of 5-15%, trichloroacetic acid concentration 30% used, to trichloroacetic acid concentration in solution be 10-20%, mix, under treating fluid normal temperature condition, the centrifugal 10min of 4000rpm, obtains clarified supernatant.
3. the method for a kind of oligopeptide content fast detecting according to claim 1, it is characterized in that it is 500nm that the described transmittance of step (2) detects wavelength, use the clarification filtration liquid that does not add trichloroacetic acid processing of step 1 acquisition as reference, as at the bottom of carrying out the sample hose after centrifugal or on wall without naked eyes visible precipitate, and use spectrophotometer to detect transmittance and reach more than 85%, can judge that the oligopeptide content in former enzymolysis liquid clear liquid exceedes 90% of Tot Prot.
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| CN201410410849.6A CN104215590A (en) | 2014-08-20 | 2014-08-20 | Method for quickly detecting oligopeptide content |
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| CN201410410849.6A CN104215590A (en) | 2014-08-20 | 2014-08-20 | Method for quickly detecting oligopeptide content |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064073A (en) * | 2016-10-27 | 2017-08-18 | 华南理工大学 | A kind of rapid assay methods of Hy drolyzed Skin Powder polypeptide molecular weight |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003042408A2 (en) * | 2001-11-12 | 2003-05-22 | University Of Wales College Of Medicine | Sequence variants of the human growth hormone gene and methods for detection |
| US7282331B2 (en) * | 2003-03-18 | 2007-10-16 | Advandx, Inc. | Method for improved specificity in probe based assays |
| CN101225106A (en) * | 2007-01-15 | 2008-07-23 | 李�昊 | Short peptide suppressing growth of cancer cell and uses thereof |
| CN103554220A (en) * | 2013-11-18 | 2014-02-05 | 长春中医药大学 | Separation and purification method of oligopeptide before and after processing nacre mother |
| CN103725742A (en) * | 2013-12-31 | 2014-04-16 | 赵树民 | Method for preparing queen bee embryo peptide by enzyme hydrolysis method |
-
2014
- 2014-08-20 CN CN201410410849.6A patent/CN104215590A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003042408A2 (en) * | 2001-11-12 | 2003-05-22 | University Of Wales College Of Medicine | Sequence variants of the human growth hormone gene and methods for detection |
| US7282331B2 (en) * | 2003-03-18 | 2007-10-16 | Advandx, Inc. | Method for improved specificity in probe based assays |
| CN101225106A (en) * | 2007-01-15 | 2008-07-23 | 李�昊 | Short peptide suppressing growth of cancer cell and uses thereof |
| CN103554220A (en) * | 2013-11-18 | 2014-02-05 | 长春中医药大学 | Separation and purification method of oligopeptide before and after processing nacre mother |
| CN103725742A (en) * | 2013-12-31 | 2014-04-16 | 赵树民 | Method for preparing queen bee embryo peptide by enzyme hydrolysis method |
Non-Patent Citations (6)
| Title |
|---|
| EIJI HASE,ET.AL: "an additional note on the nature of sulfur-containing peptide nucleotide complex obtained from chlorella and yeast", 《THE JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY》, 31 December 1959 (1959-12-31) * |
| WENSHAN HAO，HENRY K. W. FONG: "Blue and Ultraviolet Light-Absorbing Opsin from the Retinal Pigment Epithelium", 《BIOCHEMISTRY》, 31 December 1996 (1996-12-31) * |
| 任国谱等: "奶粉中低聚肽质量浓度测定方法", 《中国乳品工业》, no. 10, 31 December 2007 (2007-12-31) * |
| 罗钦等: "发酵鱼粉中寡肽检测技术研究", 《福建农业学报》, no. 03, 31 December 2008 (2008-12-31) * |
| 郭玉东等: "小肽饲料营养价值及评价方法", 《饲料工业》, no. 07, 31 December 2007 (2007-12-31) * |
| 陈升军等: "米蛋白短肽抗氧化活性研究", 《食品科技》, vol. 33, no. 12, 31 December 2008 (2008-12-31) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064073A (en) * | 2016-10-27 | 2017-08-18 | 华南理工大学 | A kind of rapid assay methods of Hy drolyzed Skin Powder polypeptide molecular weight |
| CN107064073B (en) * | 2016-10-27 | 2019-01-11 | 华南理工大学 | A kind of rapid assay methods of Hy drolyzed Skin Powder polypeptide molecular weight |
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Application publication date: 20141217 |