CN105116013A - Method for determining influence of metal ions on protease hydrolysis of soybean protein isolate - Google Patents
Method for determining influence of metal ions on protease hydrolysis of soybean protein isolate Download PDFInfo
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- CN105116013A CN105116013A CN201510195949.6A CN201510195949A CN105116013A CN 105116013 A CN105116013 A CN 105116013A CN 201510195949 A CN201510195949 A CN 201510195949A CN 105116013 A CN105116013 A CN 105116013A
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- soybean protein
- solution
- protein isolate
- thermal power
- metallic ion
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Abstract
The invention discloses a method for determining influence of metal ions on protease hydrolysis of soybean protein isolate. The method comprises the following steps: (1) preparing a solution; (2) subjecting soybean protein to water-bath treatment for 10 min; and (3) determining a thermal power-time curve. The method determines influence of the metal ions on protease hydrolysis of soybean protein isolate through research on the thermodynamic process of protease hydrolysis of soybean protein isolate, has the advantages of high sensitivity, simple operation and accurate experiment results, can provide soybean deep processing enterprises with theoretical guidance to controlling of protease hydrolysis technology applied in production of soybean peptide.
Description
Technical field
What the present invention relates to is soybean deep processing technology, especially a kind ofly determines the research method that metallic ion affects protease hydrolytic soybean protein isolate.
Background technology
Soybean protein, is a kind of high-quality albumen, has good nutrition function, but due to complex structure, causes the usable range of soybean protein and use amount to be all restricted.Demand along with soybean protein constantly increases the expansion with range of application, and its original function characteristic can not meet processing request, and this just needs to carry out modification to soybean protein, to adapt to different purposes.The modification of vegetable protein and functional study are the focuses of current developed country research.The original many functional characters of soybean protein can be made by modification etc., as dissolubility, emulsibility, holding oiliness etc. are all greatly increased, the raising of these functional characters has important value to soybean protein application in the food industry, the modification of different modes can produce different functional characteristics, thus widens the application of soybean protein.
Soybean protein forms Small molecular soybean oligopeptide through protease hydrolytic, that one has extremely strong activity and multifarious small active peptides, it does not need digestion, direct absorption also directly enters cell, play physiological activity, after it is absorbed by the body, in human body, play carrier function, other nutriments entering human body can be transported to the position that human body needs.After entering human body, play courier's effect, to neurotransmission information.There is the effects such as hypotensive, reducing blood lipid, immunity moderation function, antioxidation, subsensitivety, tranquilizing and allaying excitement.Metallic ion plays an important role in the process of protease hydrolytic soybean protein isolate, and some metallic ion plays inhibiting effect, and reduce soybean oligopeptide yield, some metallic ion plays facilitation, promotes soybean oligopeptide yield
The influence research of Chinese scholars to metallic ion determines by measuring the character of product, and test method is more complicated, and randomness is comparatively large, can not the impact effect of accurate quantitative analysis determination metallic ion.
Summary of the invention
The object of this invention is to provide a kind of method that metallic ion affects protease hydrolytic soybean protein isolate of determining, this method is by studying the thermodynamic process of protease hydrolytic soybean protein isolate, determine the impact of metallic ion on protease hydrolytic soybean protein isolate, have highly sensitive, the advantages such as simple to operate, experimental result is accurate.
Technical scheme of the present invention is: a kind ofly determine the method that metallic ion affects protease hydrolytic soybean protein isolate, comprises the following steps: (1) obtain solution:
Solution: take soybean protein sample is 10gL with the phosphate buffered solution compound concentration of different pH value
-1soy bean proteinous soln; Solution: compound concentration is 0.5gL
-1bromelain enzyme solutions; Solution: preparation is containing variable concentrations Ca
2+, Mg
2+, Li
+, Na
+metallic ion, the soy bean proteinous soln of pH=6.99.
Soybean protein carries out water bath processing 10min;
(3) measure thermal power time curve: get solution or
2ml puts into stainless steel ampoule, on the connecting rod of titration micro-calorimeter around thin plastic pipe, contain solution in plastic tube
0.5ml, first preheating, when after temperature constant, by under stirring condition, rotating speed is 120rpm, with peristaltic pump, protein enzyme solution is injected ampoule, then carries out the mensuration of thermal power-time curve;
The present invention utilizes micro-calorimeter to measure different metal ion when competitiveness is competed affects the thermal power-time curve of protease hydrolytic solution soybean protein isolate, calculates the michaelis constant of enzyme digestion reaction and maximum reaction rate respectively
The invention has the beneficial effects as follows: the present invention is by studying the thermodynamic process of protease hydrolytic soybean protein isolate, determine the impact of metallic ion on protease hydrolytic soybean protein isolate, have highly sensitive, the advantages such as simple to operate, experimental result is accurate, the protease hydrolytic technology controlling and process in can producing soybean further process enterprise Soybean Peptide well provides theoretical direction.
Accompanying drawing explanation
Fig. 1 is the thermokinetics research figure of bromelain hydrolyzate soybean protein isolate catalytic reaction.
Fig. 2 be the enzymic catalytic reaction when T=314.63K, pH=6.99 michaelis constant (
k m =50.31gL
-1) and maximum rate (
v max =0.2428gL
-1s
-1) figure.
Fig. 3 determines bromelain to the thermal power-time plot of soybean protein catalytic action when having different ions to exist and non-metallic ion exists when T=314.63K, pH=6.99.
Fig. 4 is the different metal ion (Li of variable concentrations
+, Na
+)
k 1 m with
v max, .
Fig. 5 is the different metal ion (Ca of variable concentrations
2+, Mg
2+)
k 1 m with
v max .
Embodiment:
The present invention's micro-calorimeter determines the thermal power-time curve of the enzymic catalytic reaction when T=314.63K, pH=6.99 non-metallic ion exists as Fig. 1
Ratio according to t area under a curve and the total area can obtain Reduced extent
φ, according to (5), (6) formula, parse the enzymic catalytic reaction when T=314.63K, pH=6.99 michaelis constant (
k m =50.31gL
-1) and maximum rate (
v max =0.2428gL
-1s
-1).It the results are shown in Figure 2.
Determining bromelain when T=314.63K, pH=6.99 to the thermal power-time curve of soybean protein catalytic action when having different ions to exist and non-metallic ion exists, seeing Fig. 3:
The present invention's micro-calorimeter has determined metallic ion (Li
+, Na
+) as the thermal power-time curve of competitive inhibitor when bromelain optimal reactive temperature (314.63K) and optimal reaction acidity (pH=6.99) to soybean protein catalytic action, data from thermal power-time curve, can obtain the different metal ion (Li of variable concentrations
+, Na
+)
k 1 m with
v max, its data are shown in Fig. 4.
From the data of Fig. 3, following straight-line equation can be obtained:
For inhibitor Li
+ k 1 m =0.17c+50.33R=0.9996
For inhibitor Na
+ k 1 m =0.14c+50.24R=0.9986
From research the data obtained, metallic ion Li
+, Na
+exist,
v max they be identical, and Km is different, and
k m 6 >
k m , describe metallic ion (Li
+, Na
+) be belong to Reverse transcriptase type.Suppression rule is: the Li when same concentrations
+>Na
+; When variable concentrations, suppress along with the increase of metallized metal ion concentration to increase.
Metallic ion (Ca is determined herein with micro-calorimeter
2+, Mg
2+) as competitive activator when optimal reactive temperature (314.63K) and optimal reaction acidity (pH=6.99) bromelain to the thermal power-time curve under soybean protein catalytic action.According to above-mentioned disposal route, the different metal ion (Ca of variable concentrations can be obtained
2+, Mg
2+)
k 1 m with
v max ,its data are shown in Fig. 5.
From the data of Fig. 5, following straight-line equation can be obtained:
For activator Ca
2+ k 1 m =-0.25c+50.08R=0.9953
For activator Mg
2+ k 1 m =-0.15c+50.21R=0.9980.
From research the data obtained, metallic ions Ca
2+, Mg
2+exist,
v max identical, and
k m 6 difference, but
k m >
k m 6 , describe metallic ion (Ca
2+, Mg
2+) be belong to competitive to activate type.Activation rule is: the Ca when same concentrations
2+>Mg
2+; When variable concentrations, activate increase along with the increase of concentration of metal ions.
Claims (2)
1. determine to comprise the following steps the method that metallic ion affects protease hydrolytic soybean protein isolate: (1) obtain solution:
Solution
: taking soybean protein sample, is 10gL with the phosphate buffered solution compound concentration of different pH value
-1soy bean proteinous soln; Solution
: compound concentration is 0.5gL
-1bromelain enzyme solutions; Solution
: preparation is containing variable concentrations Ca
2+, Mg
2+, Li
+, Na
+metallic ion, the soy bean proteinous soln of pH=6.99.
2. (2) soybean protein carries out water bath processing 10min;
(3) thermal power time curve is measured: get solution
or
2ml puts into stainless steel ampoule, on the connecting rod of titration micro-calorimeter around thin plastic pipe, contain solution in plastic tube
0.5ml, first preheating, when after temperature constant, by under stirring condition, rotating speed is 120rpm, with peristaltic pump protein enzyme solution
inject ampoule, then carry out the mensuration of thermal power-time curve;
A kind of method that metallic ion affects protease hydrolytic soybean protein isolate of determining according to claim 1, it is characterized in that: utilizing micro-calorimeter to measure different metal ion when competitiveness is competed affects the thermal power-time curve of protease hydrolytic solution soybean protein isolate, calculates the michaelis constant of enzyme digestion reaction and maximum reaction rate respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510195949.6A CN105116013A (en) | 2015-04-23 | 2015-04-23 | Method for determining influence of metal ions on protease hydrolysis of soybean protein isolate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510195949.6A CN105116013A (en) | 2015-04-23 | 2015-04-23 | Method for determining influence of metal ions on protease hydrolysis of soybean protein isolate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105116013A true CN105116013A (en) | 2015-12-02 |
Family
ID=54664053
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510195949.6A Pending CN105116013A (en) | 2015-04-23 | 2015-04-23 | Method for determining influence of metal ions on protease hydrolysis of soybean protein isolate |
Country Status (1)
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001034660A2 (en) * | 1999-11-09 | 2001-05-17 | Sri International | Screening and analysis of polymers, specialty chemicals and catalysts using radiography |
| CN2519909Y (en) * | 2001-09-26 | 2002-11-06 | 上海理工大学 | Heat-measuring type biosensor for detecting residul pesticide |
| CN101351702A (en) * | 2005-12-07 | 2009-01-21 | 惠氏公司 | Methods for preparing crystalline rapamycin and for measuring crystallinity of rapamycin compounds using differential scanning calorimetry |
| CN102980912A (en) * | 2012-11-13 | 2013-03-20 | 湖北中烟工业有限责任公司 | Bio-thermo-chemical method used in cigarette flavor and fragrance security screening |
| CN103196952A (en) * | 2013-04-07 | 2013-07-10 | 青岛大学 | Calorimetric method for detecting pesticides by using acetylcholin esterase |
-
2015
- 2015-04-23 CN CN201510195949.6A patent/CN105116013A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001034660A2 (en) * | 1999-11-09 | 2001-05-17 | Sri International | Screening and analysis of polymers, specialty chemicals and catalysts using radiography |
| CN2519909Y (en) * | 2001-09-26 | 2002-11-06 | 上海理工大学 | Heat-measuring type biosensor for detecting residul pesticide |
| CN101351702A (en) * | 2005-12-07 | 2009-01-21 | 惠氏公司 | Methods for preparing crystalline rapamycin and for measuring crystallinity of rapamycin compounds using differential scanning calorimetry |
| CN102980912A (en) * | 2012-11-13 | 2013-03-20 | 湖北中烟工业有限责任公司 | Bio-thermo-chemical method used in cigarette flavor and fragrance security screening |
| CN103196952A (en) * | 2013-04-07 | 2013-07-10 | 青岛大学 | Calorimetric method for detecting pesticides by using acetylcholin esterase |
Non-Patent Citations (1)
| Title |
|---|
| 李向阳等: "菠萝蛋白酶催化大豆蛋白水解反应的热动力学", 《高等学校化学学报》 * |
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| PB01 | Publication | ||
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Application publication date: 20151202 |