CN105728036B - Bovine serum albumin(BSA) platinum/bismuth composite nano materials Mimetic enzyme - Google Patents
Bovine serum albumin(BSA) platinum/bismuth composite nano materials Mimetic enzyme Download PDFInfo
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Abstract
The present invention discloses a kind of bovine serum albumin(BSA) platinum/bismuth composite nano materials Mimetic enzyme, aqueous bismuth nitrate solution is added in the bovine serum albumin(BSA) platinum nano material aqueous solution, heating water bath after being mixed with phosphate buffer, the mixed solution after heating obtain bismuth bovine serum albumin(BSA) platinum nano composite material Mimetic enzyme by ultrafiltration and washing.Bovine serum albumin(BSA) platinum/bismuth composite nano materials simulation peroxidase can catalyzing hydrogen peroxide oxidation 3,3 ', 5,5 ' tetramethyl biphenyl amine hydrochlorates develop the color, to 3,3 ', 5, the Michaelis constant of 5 ' tetramethyl biphenyl amine hydrochlorates is 0.054 mmol/L, and the Michaelis constant to hydrogen peroxide is 13.24 mmol/L.The analogue enztme acid and alkali-resistance, high temperature resistant, resistance to high salt simultaneously, have excellent room temperature stability.
Description
Technical field
The present invention relates to bovine serum albumin(BSA)-platinum/bismuth composite nano materials with Mimetic enzyme characteristic, belong to
Nanometer technology and bionics techniques field.
Background technology
Enzyme is a kind of protein, RNA or complex in vivo with catalytic activity.The selectivity of enzymatic,
High efficiency and Modulatory character determine that it is as the core substance that catalysis substrate is reacted in life entity.The application of enzyme is very
Extensively, it can not only be provided using various biology enzymes for field of scientific study and greatly help also largely to improve
The quality of life or even the general level of the health of people.But enzyme is easily influenceed and lost activity by a variety of physics, chemical factor, and
And the limited source of native enzyme, expensive, accuracy and fineness to experimental implementation have very high requirement.Therefore, with
Research is goed deep into, and the synthesis and use to various artificial mimic enzymes are as the focus studied.
Peroxidase is a kind of oxidoreducing enzyme as caused by microorganism or plant, and they can be catalyzed many reactions,
And substrate oxidation is catalyzed by electron acceptor of hydrogen peroxide, and hydrogen peroxide is intermediate product important in biochemical reaction, thing
The existence of kind be unable to do without the catalytic reaction that hydrogen peroxide participates in.Natural peroxide enzyme content is few, easily by external environmental interference, valency
Lattice are expensive.Therefore the research of the novel nano-material for possessing Mimetic enzyme activity is increasingly closed by people
Note.
The invention provides a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials mould with high activity, high stability
Intend peroxidase.
The content of the invention
It is an object of the invention to provide a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme and
Its preparation method.
To achieve these goals, the present invention uses following technical scheme：
Bovine serum albumin(BSA)-platinum of the present invention/bismuth composite nano materials Mimetic enzyme, it is characterized in that under
What the method for stating was prepared：It is 16 to add 5 mL concentration in 5 mL concentration are 50 mg/mL Bovine Serum Albumin in Aqueous Solution
Mmol/L platinum acid chloride solutions and 0.5 mL concentration are 1.5 mol/L sodium hydroxide solutions, react 2 h at 80 DEG C of water-bath after mixing,
Solution loads the super filter tube that cutoff is 3k, 6000 r/min centrifugal ultrafiltrations after reaction, and washes 3 times, obtains cow's serum
Albumin-platinum nano composite material the aqueous solution, bovine serum albumin(BSA)-platinum nano composite material aqueous solution is freeze-dried to obtain ox
The powder of seralbumin-platinum nano composite material；In bovine serum albumin(BSA)-platinum nanometer that 0.6 mL concentration is 23.8 mg/mL
It is 10 mmol/ that bismuth nitrate solution that 0.6 mL concentration is 0.5 mmol/L and 1.8 mL concentration are added in the composite aqueous solution
L, the phosphate buffer of pH=7,2.5 h are reacted after mixing at 80 DEG C of water-bath, it is 3k that solution, which loads cutoff, after reaction
Super filter tube, 6000 r/min centrifugal ultrafiltrations, and wash 3 times, obtain the water of bovine serum albumin(BSA)-platinum/bismuth composite nano materials
Solution.
Described a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that in phosphoric acid
Hydrogen peroxide is sequentially added in salt buffer, TMB hydrochloride and bovine serum albumin(BSA)-platinum/bismuth are multiple
The nano material aqueous solution, warm bath after mixing are closed, solution is changed into blueness from colourless, there is a maximum absorption band at 652 nm.
Described a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that being catalyzed
The activity of oxidation hydroxide 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate reaches maximum in pH=5.
Described a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that being catalyzed
The activity of oxidation hydroxide 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate reaches maximum at 37 DEG C.
A kind of described bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that to 3,
3 ', 5, the Michaelis constant of 5 '-tetramethyl biphenyl amine hydrochlorate is 0.054 mmol/L, and the Michaelis constant to hydrogen peroxide is
13.24 mmol/L。
Described a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that 2.79
ML concentration is 100 mmol/L, and the hydrogen peroxide that 1 mL concentration is 2 mol/L is sequentially added in the phosphate buffer of pH=5,
0.2 mL concentration is 16 mmol/L TMB hydrochloride, and 10 μ L concentration are 4.76 mg/mL ox
Seralbumin-platinum/bismuth composite nano materials the aqueous solution, 37 DEG C of warm bath 10 minutes after mixing, solution are changed into blueness from colourless,
There is a maximum absorption band at 652 nm.
A kind of described bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that 30 ~
Peroxidase activity is simulated after being preserved 2 hours at a temperature of 80 DEG C without significant change, there is good stability.
Described a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that in pH 3
Peroxidase activity is simulated without significant change after ~ 11 preservations 2 hours, and there is good stability.
A kind of described bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that 0 ~
Peroxidase activity is simulated after being preserved 2 hours in 7.5 mol/L sodium chloride solutions without significant change, there is good stabilization
Property；Or activity without significant change, has good room temperature stability, and relative catalytic activity reaches after preserving 30 days at room temperature
To 98.9%.
A kind of preparation side of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme of the present invention
Method, comprise the following steps：It is 16 to add 5 mL concentration in 5 mL concentration are 50 mg/mL Bovine Serum Albumin in Aqueous Solution
Mmol/L platinum acid chloride solutions and 0.5 mL concentration are 1.5 mol/L sodium hydroxide solutions, react 2 h at 80 DEG C of water-bath after mixing,
Solution loads the super filter tube that cutoff is 3k, 6000 r/min centrifugal ultrafiltrations after reaction, and washes 3 times, obtains cow's serum
Albumin-platinum nano composite material the aqueous solution, bovine serum albumin(BSA)-platinum nano composite material aqueous solution is freeze-dried to obtain ox
The powder of seralbumin-platinum nano composite material；In bovine serum albumin(BSA)-platinum nanometer that 0.6 mL concentration is 23.8 mg/mL
It is 10 mmol/ that bismuth nitrate solution that 0.6 mL concentration is 0.5 mmol/L and 1.8 mL concentration are added in the composite aqueous solution
L, the phosphate buffer of pH=7,2.5 h are reacted after mixing at 80 DEG C of water-bath, it is 3k that solution, which loads cutoff, after reaction
Super filter tube, 6000 r/min centrifugal ultrafiltrations, and wash 3 times, obtain the water of bovine serum albumin(BSA)-platinum/bismuth composite nano materials
Solution.
Specifically,（One）A kind of bovine serum albumin(BSA)-platinum of the present invention/bismuth composite nano materials simulation peroxidating
Thing enzyme, it is prepared by following methods：It is water-soluble that bismuth nitrate is added in the aqueous solution of bovine serum albumin(BSA)-platinum nano material
Liquid, heating water bath after mixing, ultrafiltration are simultaneously washed, and obtain bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution.
The proportioning of each component in the preparation of bovine serum albumin(BSA)-platinum/bismuth composite nano materials of the present invention is by this
What field conventional method can be completed.But the preparation of currently preferred bovine serum albumin(BSA)-platinum/bismuth composite nano materials by with
It is prepared by lower section method：0.6 mL concentration is added in the bovine serum albumin(BSA) for being 23.8 mg/mL in 0.6 mL concentration-platinum aqueous solution is
0.5 mmol/L bismuth nitrate solution and 1.8 mL concentration are 10 mmol/L, the phosphate buffer of pH=7, water-bath 80 after mixing
2.5 h are reacted at DEG C.Solution loads the super filter tube that cutoff is 3k, 6000 r/min centrifugal ultrafiltrations after reaction, and washes 3
It is secondary, obtain the aqueous solution of bovine serum albumin(BSA)-platinum/bismuth composite nano materials.
Described bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution obtains bovine serum albumin by freeze-drying
In vain-platinum/bismuth composite nano materials powder.All glasswares used in above procedure soak by chloroazotic acid, and are steamed with double
Water thoroughly cleans, and dries.
（Two）The peroxidase activity of bovine serum albumin(BSA)-platinum/bismuth composite nano materials of the present invention
Hydrogen peroxide, 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate and cow's serum are sequentially added in phosphate buffer
Albumin-platinum/bismuth composite nano materials the aqueous solution, warm bath 10 minutes after mixing, observe color change and determine UV, visible light suction
Receive spectrum.
Bovine serum albumin(BSA)-platinum of the present invention/bismuth composite nano materials Mimetic enzyme, can be catalyzed peroxidating
Hydroxide TMB hydrochloride generates blue product, and the product has maximum absorption band at 652 nm.
Bovine serum albumin(BSA)-platinum of the present invention/bismuth composite nano materials Mimetic enzyme catalyzing hydrogen peroxide
The activity of oxidation TMB hydrochloride reaches maximum in pH=5,37 DEG C, and its peroxidase
Activity is significantly improved compared with bovine serum albumin(BSA)-platinum nano material.
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme of the present invention to 3,3 ', 5,5 '-
The Michaelis constant of tetramethyl biphenyl amine hydrochlorate is 0.054 mmol/L, and the Michaelis constant to hydrogen peroxide is 13.24 mmol/
L。
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme of the present invention is in 30 ~ 80 DEG C of temperature
Catalytic activity is without obvious change after degree preserves 2 hours either in pH 3 ~ 11 or in concentration being 0 ~ 7.5 M sodium chloride solutions
Change, there is good stability.
Bovine serum albumin(BSA)-platinum of the present invention/bismuth composite nano materials Mimetic enzyme has good room
Temperature stability, after preserving 30 days at room temperature, relative catalytic activity can keep 98.9%.
Advantages of the present invention：
（1）Preparation method of the present invention is green, easy to operate quick, it is not necessary to use reducing agent.
（2）There is bovine serum albumin(BSA)-platinum/bismuth composite nano materials in the present invention good Mimetic enzyme to live
Property and its peroxidase activity is significantly improved compared with bovine serum albumin(BSA)-platinum nano material.
（3）Bovine serum albumin(BSA)-platinum/bismuth composite nano materials stability in the present invention is good, peroxidase activity by
The influence of storage temperature, pH value and ionic strength is small.
（4）Bovine serum albumin(BSA)-platinum/bismuth composite nano materials in the present invention have good room temperature long-time stability,
After preserving 30 days at room temperature, relative catalytic activity can keep 98.9%.
Brief description of the drawings
Fig. 1 is that bovine serum albumin(BSA)-platinum/bismuth composite nano materials are catalyzed peroxide with bovine serum albumin(BSA)-platinum nano material
Change the ultra-violet absorption spectrum comparison diagram of hydroxide 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate color development system.A is blank in figure
Group, b are bovine serum albumin(BSA)-platinum nano material group, and c is bovine serum albumin(BSA)-platinum/bismuth composite nano materials group.
Fig. 2 is pH value to bovine serum albumin(BSA)-platinum/bismuth composite nano materials catalyzing hydrogen peroxide oxidation 3,3 ', 5,5 '-four
The influence figure of methyl biphenyl amine hydrochlorate color development system.
Fig. 3 is that warm bath temperature aoxidizes 3,3 ', 5 to bovine serum albumin(BSA)-platinum/bismuth composite nano materials catalyzing hydrogen peroxide,
The influence figure of 5 '-tetramethyl biphenyl amine hydrochlorate color development system.
Fig. 4 is that bovine serum albumin(BSA)-platinum/bismuth composite nano materials are steady to 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate
State dynamic curve diagram.
Fig. 5 is stability kinetics curve map of the bovine serum albumin(BSA)-platinum/bismuth composite nano materials to hydrogen peroxide.
Fig. 6 is the pH stability diagrams of bovine serum albumin(BSA)-platinum/bismuth composite nano materials.
Fig. 7 is the ionic strength stability diagram of bovine serum albumin(BSA)-platinum/bismuth composite nano materials.
Fig. 8 is the temperature stability figure of bovine serum albumin(BSA)-platinum/bismuth composite nano materials.
Embodiment
Example 1：
It is 16 mmol/L chlorine to add 5 mL concentration in 5 mL concentration are 50 mg/mL Bovine Serum Albumin in Aqueous Solution
Platinic acid solution and 0.5 mL concentration are 1.5 mol/L sodium hydroxide solutions, react 2 h at 80 DEG C of water-bath after mixing.It is molten after reaction
Liquid loads the super filter tube that cutoff is 3k, 6000 r/min centrifugal ultrafiltrations, and washes 3 times, obtains bovine serum albumin(BSA)-platinum
The nano composite material aqueous solution, bovine serum albumin(BSA)-platinum nano composite material aqueous solution is freeze-dried to obtain bovine serum albumin
The powder of in vain-platinum nano composite material.Added in bovine serum albumin(BSA)-platinum aqueous solution that 0.6 mL concentration is 23.8 mg/mL
The bismuth nitrate solution and 1.8 mL concentration that 0.6 mL concentration is 0.5 mmol/L are 10 mmol/L, the phosphate-buffered of pH=7
Liquid, 2.5 h are reacted after mixing at 80 DEG C of water-bath, solution loads the super filter tube that cutoff is 3k, 6000 r/min after reaction
Centrifugal ultrafiltration, and wash 3 times, obtain bovine serum albumin(BSA)-platinum/bismuth composite nano materials（Also referred to as：Bismuth-bovine serum albumin(BSA)-
Platinum nano composite material）The aqueous solution.
Example 2：
It is 100 mmol/L in 2.79 mL concentration, 1 mL concentration is sequentially added in the phosphate buffer of pH=5 as 2
Mol/L hydrogen peroxide, 0.2 mL concentration be 16 mmol/L TMB hydrochloride, 10 μ L concentration
Bovine serum albumin(BSA)-platinum/bismuth the composite nano materials or bovine serum albumin(BSA)-platinum nanometer prepared for 4.76 mg/mL example 1
The material aqueous solution, 37 DEG C of warm bath 10 minutes after mixing, determines ultraviolet-visible absorption spectroscopy.As shown in figure 1, bovine serum albumin(BSA)-
Platinum/bismuth composite nano materials and bovine serum albumin(BSA)-platinum nano material can be catalyzed 3,3 ', 5,5 '-tetramethyl benzidine hydrochloric acid
Salt develops the color, and has a maximum absorption band, bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme at 652 nm
Activity is apparently higher than bovine serum albumin(BSA)-platinum nano material.
Example 3：
It is 100 mmol/L in 2.79 mL concentration, different pH value（2-10）Phosphate buffer in sequentially add 1 mL
Concentration is 2 mol/L hydrogen peroxide, and 0.2 mL concentration is 16 mmol/L TMB hydrochloride,
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution prepared by the example 1 that 10 μ L concentration are 4.76 mg/mL, after mixing
37 DEG C of warm bath 10 minutes, determine absorbance at 652 nm wavelength.As shown in Fig. 2 bovine serum albumin(BSA)-platinum/bismuth composite Nano material
The Mimetic enzyme activity of material reaches maximum in pH=5.
Example 4：
It is 100 mmol/L in 2.79 mL concentration, 1 mL concentration is sequentially added in the phosphate buffer of pH=5 as 2
Mol/L hydrogen peroxide, 0.2 mL concentration be 16 mmol/L TMB hydrochloride, 10 μ L concentration
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials the aqueous solution prepared for 4.76 mg/mL example 1, different after mixing
Temperature（20-70 ℃）Lower warm bath 10 minutes, determine absorbance at 652 nm wavelength.As shown in figure 3, bovine serum albumin(BSA)-platinum/
The Mimetic enzyme activity of bismuth composite nano materials reaches maximum at 37 DEG C.
Example 5：
It is 100 mmol/L in 2.79 mL concentration, 1 mL concentration is sequentially added in the phosphate buffer of pH=5 as 2
Mol/L hydrogen peroxide, 0.2 mL various concentrations（0.025-0.5 mmol/L）3,3 ', 5,5 '-tetramethyl benzidine hydrochloric acid
Salt, bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution prepared by the example 1 that 10 μ L concentration are 4.76 mg/mL, is mixed
37 DEG C of warm bath 1 minute after conjunction, determine the absorbance at 652 nm wavelength, are fitted according to Michaelis-Menten equation.As shown in figure 4,
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials are 0.054 mmol/L to TMB Michaelis constant.
Example 6：
It is 100 mmol/L in 2.79 mL concentration, 1 mL various concentrations is sequentially added in the phosphate buffer of pH=5
（0.0045-0.35 mol/L）Hydrogen peroxide, 0.2 mL concentration be 16 mmol/L TMB salt
Hydrochlorate, bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution prepared by the example 1 that 10 μ L concentration are 4.76 mg/mL,
37 DEG C of warm bath 1 minute after mixing, determine the absorbance at 652 nm wavelength, are fitted according to Michaelis-Menten equation.Such as Fig. 5 institutes
Show, bovine serum albumin(BSA)-platinum/bismuth composite nano materials are 13.24 mmol/L to the Michaelis constant of hydrogen peroxide.
Example 7：
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution is placed under condition of different pH after preserving 2 hours, surveyed
Surely influences of the pH to its Mimetic enzyme activity is preserved.It is 100 mmol/L in 2.79 mL concentration, the phosphate of pH=5 delays
The hydrogen peroxide that 1 mL concentration is 2 mol/L is sequentially added in fliud flushing, 0.2 mL concentration is 3,3 ', 5, the 5 '-four of 16 mmol/L
Methyl biphenyl amine hydrochlorate, bovine serum albumin(BSA)-platinum/bismuth composite Nano prepared by the example 1 that 10 μ L concentration are 4.76 mg/mL
The material aqueous solution, 37 DEG C of warm bath 10 minutes after mixing, determines the absorbance at 652 nm wavelength.As shown in fig. 6, cow's serum
Albumin-platinum/bismuth composite nano materials simulate peroxidase activity without bright after being preserved 2 hours under conditions of pH 3 ~ 11
Aobvious change, has good stability.
Example 8：
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution is placed in various concentrations sodium chloride and preserved 2 hours
Afterwards, influence of the sodium chloride concentration to its Mimetic enzyme activity is determined.It is 100 mmol/L in 2.79 mL concentration, pH=5
Phosphate buffer in sequentially add 1 mL concentration be 2 mol/L hydrogen peroxide, 0.2 mL concentration be 16 mmol/L 3,
3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate, bovine serum albumin(BSA)-platinum prepared by the example 1 that 10 μ L concentration are 4.76 mg/mL/
The bismuth composite nano materials aqueous solution, 37 DEG C of warm bath 10 minutes after mixing, determines the absorbance at 652 nm wavelength.Such as Fig. 7 institutes
Show, bovine serum albumin(BSA)-platinum/bismuth composite nano materials simulate peroxidating after being preserved 2 hours in 0 ~ 7.5 mol/L sodium chloride
Thing enzymatic activity has no significant change, and has good stability.
Example 9：
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution is placed under condition of different temperatures after preserving 2 hours,
Determine influence of the storage temperature to its Mimetic enzyme activity.It is 100 mmol/L in 2.79 mL concentration, the phosphoric acid of pH=5
The hydrogen peroxide that 1 mL concentration is 2 mol/L is sequentially added in salt buffer, 0.2 mL concentration is the 3,3 ', 5 of 16 mmol/L,
5 '-tetramethyl biphenyl amine hydrochlorate, bovine serum albumin(BSA)-platinum/bismuth prepared by the example 1 that 10 μ L concentration are 4.76 mg/mL are multiple
The nano material aqueous solution is closed, 37 DEG C of warm bath 10 minutes after mixing, determines the absorbance at 652 nm wavelength.As shown in figure 8,
Mimetic enzyme after bovine serum albumin(BSA)-platinum/bismuth composite nano materials preserve 2 hours under 30 ~ 80 DEG C of temperature conditionss
Activity has no significant change, and has good stability.
Example 10：
Bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution is placed in and preserved 30 days at room temperature, it is determined and simulated
Peroxidase activity.It is 100 mmol/L in 2.79 mL concentration, sequentially adding 1 mL concentration in the phosphate buffer of pH=5 is
2 mol/L hydrogen peroxide, 0.2 mL concentration are 16 mmol/L TMB hydrochloride, and 10 μ L are dense
Spend bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution of the preparation of example 1 for 4.76 mg/mL, 37 DEG C of temperature after mixing
Bath 10 minutes, determine the absorbance at 652 nm wavelength.As a result shown bovine serum albumin(BSA)-platinum/bismuth composite nano materials exist
Its relative catalytic activity can reach 98.9% after room temperature placed 30 days.
The exemplary embodiments of the present invention are the foregoing is only, are not intended to limit the invention, all essences in the present invention
God and any modification made within principle, equivalent substitution and improvement etc., should be included in the scope of the protection.
Claims (10)
- A kind of 1. bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, it is characterized in that being prepared by following methods Form：It is 16 mmol/L chlorine platinum that 5 mL concentration are added in 5 mL concentration are 50 mg/mL Bovine Serum Albumin in Aqueous Solution Acid solution and 0.5 mL concentration are 1.5 mol/L sodium hydroxide solutions, react 2 h, solution after reaction after mixing at 80 DEG C of water-bath Load the super filter tube that cutoff is 3k, 6000 r/min centrifugal ultrafiltrations, and wash 3 times, obtain bovine serum albumin(BSA)-platinum and receive The nano composite material aqueous solution, by bovine serum albumin(BSA)-platinum nano composite material aqueous solution be freeze-dried to obtain bovine serum albumin(BSA)- The powder of platinum nano composite material；In bovine serum albumin(BSA)-platinum nano composite material water that 0.6 mL concentration is 23.8 mg/mL Add bismuth nitrate solution that 0.6 mL concentration is 0.5 mmol/L in solution and 1.8 mL concentration are 10 mmol/L, the phosphorus of pH=7 Phthalate buffer, 2.5 h reacting after mixing at 80 DEG C of water-bath, solution loads the super filter tube that cutoff is 3k after reaction, 6000 r/min centrifugal ultrafiltrations, and wash 3 times, obtain the aqueous solution of bovine serum albumin(BSA)-platinum/bismuth composite nano materials.
- 2. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1, its It is characterized in sequentially adding hydrogen peroxide in phosphate buffer, TMB hydrochloride and ox blood are pure Albumen-platinum/bismuth composite nano materials the aqueous solution, warm bath after mixing, solution are changed into blueness from colourless, there is a maximum at 652 nm Absworption peak.
- 3. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1 or 2, It is characterized in that the activity of catalyzing hydrogen peroxide oxidation 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate reaches maximum in pH=5.
- 4. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1 or 2, It is characterized in that the activity of catalyzing hydrogen peroxide oxidation 3,3 ', 5,5 '-tetramethyl biphenyl amine hydrochlorate reaches maximum at 37 DEG C.
- 5. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1 or 2, It is characterized in that the Michaelis constant to TMB hydrochloride is 0.054 mmol/L, to the rice of hydrogen peroxide Family name's constant is 13.24 mmol/L.
- 6. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1 or 2, It is characterized in that being 100 mmol/L in 2.79 mL concentration, 1 mL concentration is sequentially added in the phosphate buffer of pH=5 as 2 Mol/L hydrogen peroxide, 0.2 mL concentration be 16 mmol/L TMB hydrochloride, 10 μ L concentration For 4.76 mg/mL bovine serum albumin(BSA)-platinum/bismuth composite nano materials aqueous solution, 37 DEG C of warm bath 10 minutes, solution after mixing It is changed into blueness from colourless, there is a maximum absorption band at 652 nm.
- 7. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1, its It is characterized in simulating peroxidase activity at a temperature of 30 ~ 80 DEG C after preserving 2 hours without significant change, there is good stabilization Property.
- 8. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1, its It is characterized in simulating peroxidase activity without significant change after pH 3 ~ 11 is preserved 2 hours, there is good stability.
- 9. a kind of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme according to claim 1, its It is characterized in simulating peroxidase activity in 0 ~ 7.5 mol/L sodium chloride solutions after preserving 2 hours without significant change, has good Good stability；Or activity without significant change, has good room temperature stability, and urge relatively after preserving 30 days at room temperature Change activity and reach 98.9%.
- 10. a kind of preparation method of bovine serum albumin(BSA)-platinum/bismuth composite nano materials Mimetic enzyme, including following step Suddenly：It is that 16 mmol/L chloroplatinic acids are molten to add 5 mL concentration in 5 mL concentration are 50 mg/mL Bovine Serum Albumin in Aqueous Solution Liquid and 0.5 mL concentration are 1.5 mol/L sodium hydroxide solutions, react 2 h after mixing at 80 DEG C of water-bath, and solution loads after reaction Cutoff be 3k super filter tube, 6000 r/min centrifugal ultrafiltrations, and wash 3 times, obtain bovine serum albumin(BSA)-platinum nanometer and answer The condensation material aqueous solution, bovine serum albumin(BSA)-platinum nano composite material aqueous solution is freeze-dried to obtain bovine serum albumin(BSA)-platinum and received The powder of nano composite material；In bovine serum albumin(BSA)-platinum nano composite material aqueous solution that 0.6 mL concentration is 23.8 mg/mL The middle bismuth nitrate solution that 0.6 mL concentration is 0.5 mmol/L and 1.8 mL concentration of adding is 10 mmol/L, the phosphate of pH=7 Buffer solution, 2.5 h are reacted after mixing at 80 DEG C of water-bath, the super filter tube that solution loading cutoff is 3k after reaction, 6000 R/min centrifugal ultrafiltrations, and wash 3 times, obtain the aqueous solution of bovine serum albumin(BSA)-platinum/bismuth composite nano materials.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610063490.9A CN105728036B (en) | 2016-01-29 | 2016-01-29 | Bovine serum albumin(BSA) platinum/bismuth composite nano materials Mimetic enzyme |
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| CN201610063490.9A CN105728036B (en) | 2016-01-29 | 2016-01-29 | Bovine serum albumin(BSA) platinum/bismuth composite nano materials Mimetic enzyme |
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| "贵金属纳米材料模拟过氧化物酶的应用研究";陈丰庆;《海峡药学》;20151231;第27卷(第5期);第5-8页 * |
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