CN110243897B - Selective recognition of tryptophan enantiomer by polyethyleneimine/chiral peptide modified electrode - Google Patents
Selective recognition of tryptophan enantiomer by polyethyleneimine/chiral peptide modified electrode Download PDFInfo
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Abstract
The invention relates to a preparation method of a polyethyleneimine/chiral peptide modified glassy carbon electrode and application of the polyethyleneimine/chiral peptide modified glassy carbon electrode in selective recognition of tryptophan enantiomer. The method comprises the following steps: preparing a polyethyleneimine modified glassy carbon electrode, and then soaking the polyethyleneimine modified glassy carbon electrode into a chiral peptide (BGAc) solution for further modification to obtain the polyethyleneimine/chiral peptide modified glassy carbon electrode for electrochemical recognition of tryptophan enantiomers. The invention has the following effects: the preparation process of the polyethyleneimine/chiral peptide modified electrode is simple, environment-friendly and low in cost, and the modified electrode has an obvious identification effect on tryptophan enantiomers.
Description
Technical Field
The invention relates to preparation of a polyethyleneimine/chiral peptide modified glassy carbon electrode and application of the polyethyleneimine/chiral peptide modified glassy carbon electrode to selective recognition of tryptophan enantiomers, and belongs to the field of electrochemical analysis and biotechnology.
Background
The amino acid provides an important material basis for the metabolism, the life activity and the growth and development of organisms, and can be used as an important biomarker for various metabolic diseases. The amino acid isomer configuration has D type and L type, and in practical application, one enantiomer has pharmacological action on human body, while the other enantiomer has no effect or serious side effect or even toxic effect. Therefore, the identification research of the chiral amino acid is particularly important and significant in the fields of pharmacy, biological analysis, drug screening, analysis and separation and the like. The existing spectrum analysis methods have the defects of low sensitivity, complex operation, high analysis cost and the like, so that the search for an analysis method with good prospect is of great importance. The electrochemical analysis method has the advantages of high sensitivity, quick response time, simple and convenient operation, and the like, and can distinguish different amino acid enantiomers by regulating and controlling the chiral interface of the modified electrode, so the electrochemical detection based on chiral recognition has better application prospect.
Disclosure of Invention
In the invention, the polyethyleneimine and the chiral peptide are self-assembled to form a film on the glassy carbon electrode by utilizing the electrostatic action of the polyethyleneimine and the chiral peptide. By means of the good film forming capability of polyethyleneimine, the chiral peptide region is combined with tryptophan enantiomer.
The preparation method of the polyethyleneimine/chiral peptide modified glassy carbon electrode and the application of the polyethyleneimine/chiral peptide modified glassy carbon electrode in the selective recognition of tryptophan enantiomers comprises the following steps:
a. preparing a polyethyleneimine modified electrode: preparing a polyethyleneimine solution, and immersing the glassy carbon electrode in the polyethyleneimine solution for a certain time to obtain a polyethyleneimine modified electrode;
b. preparing a polyethyleneimine/chiral peptide modified electrode: preparing a chiral peptide solution, washing a polyethyleneimine modified electrode with water, immersing the electrode into the chiral peptide solution for a certain time, taking out the electrode, and drying the electrode at room temperature to obtain the polyethyleneimine/chiral peptide modified electrode;
c. electrochemical identification of tryptophan enantiomers: the polyethyleneimine/chiral peptide modified electrode is a working electrode, the Ag/AgCl electrode is a reference electrode, the platinum wire electrode is a counter electrode, the three-electrode system is immersed in the prepared L-tryptophan or D-tryptophan solution, and after incubation for a certain time, the test is carried out by using a pulse voltammetry method.
Further, in the step a, soaking the water solution with the polyethyleneimine concentration of 1-3 mg/mL for 20-60 min.
Further, in the step b, the chiral peptide is BGAc, the concentration of the chiral peptide is 0.1-1 mg/mL of aqueous solution, and the soaking time is 10-30 min.
The BGAc has a structural formula as follows:
furthermore, the concentration of the L-/D-tryptophan solution in the step c is 0.01-0.8 mM, the scanning potential is 0.4-1V, and the incubation time is 1-30 min.
The invention has the following beneficial effects:
according to the invention, polyethyleneimine and chiral peptide are self-assembled to form a film on a glassy carbon electrode as a chiral amino acid recognition material, compared with a single polyethyleneimine or chiral peptide modified electrode, the composite film material modified electrode has the advantages that the recognition effect of tryptophan enantiomer is obviously improved, and the preparation method is cheap, environment-friendly and simple.
Drawings
FIG. 1 is a scanning electron microscope image of a Polyethyleneimine (PEI) membrane (A), a chiral peptide (D-BGAc) (B) and a PEI/D-BGAc composite membrane (C).
FIG. 2 is a DPV graph of L-/D-tryptophan enantiomer recognition by bare glass carbon electrodes.
FIG. 3 is a DPV diagram of recognition of L-/D-tryptophan enantiomer by PEI modified glassy carbon electrode.
FIG. 4 is a DPV graph of recognition of the L-/D-tryptophan enantiomer by a chiral peptide (D-BGAc) modified glassy carbon electrode.
FIG. 5 is a DPV diagram of recognition of the L-/D-tryptophan enantiomer by PEI/D-BGAc vitrifying carbon electrodes.
FIG. 6 is a graph showing the relationship between the content of different L-tryptophan in a mixed solution and the current measured by a PEI/D-BGAc modified glassy carbon electrode.
Detailed Description
The invention will now be further described with reference to specific embodiments, which are intended to illustrate but not to limit the invention further.
The first embodiment is as follows:
preparation of PEI/D-BGAc for characterization of scanning electron microscope
a. 50mg PEI was weighed and dissolved in 50mL deionized water to make a 1mg/mL PEI solution.
b. Method reference [1] for preparing D-BGAc by weighing 25mgD-BGAc chiral peptide, and dispersing in 50mL deionized water to obtain 0.5mg/mLD-BGAc solution.
c. And cleaning the silicon wafer, blowing the silicon wafer to dry by using nitrogen, placing the silicon wafer into a 1mg/mL PEI solution, and soaking for 15min to obtain the PEI modified silicon wafer.
d. And D, cleaning the PEI modified silicon wafer prepared in the step c by using deionized water, drying by blowing nitrogen, immersing into a D-BGAc solution containing 0.5mg/mL, soaking for 15min, taking out, and airing at room temperature to obtain the PEI/D-BGAc membrane.
Example two:
the steps for respectively preparing the PEI modified electrode, the D-BGAc modified electrode and the PEI/D-BGAc modified glassy carbon electrode for electrochemical recognition of tryptophan enantiomer are as follows:
preparing a PEI modified electrode: preparing 4mL of 1mg/mLPEI solution, immersing the glassy carbon electrode into the PEI solution, and immersing for 30min to obtain the PEI modified glassy carbon electrode.
Preparing a D-BGAc modified electrode: preparing 4mL of 0.5mg/mLD-BGAc solution, immersing the glassy carbon electrode into the D-BGAc solution, and soaking for 30min to obtain the D-BGAc modified glassy carbon electrode.
Preparing a PEI/D-BGAc modified electrode: and cleaning the prepared PEI modified glassy carbon electrode with deionized water, drying with nitrogen, soaking in a D-BGAc solution containing 0.5mg/mL, and soaking for 15min to obtain the PEI/D-BGAc modified glassy carbon electrode.
And (3) taking the PEI modified electrode, the D-BGAc modified electrode or the PEI/D-BGAc modified glassy carbon prepared in the above step as a working electrode, an Ag/AgCl electrode as a reference electrode, a platinum wire electrode as a counter electrode, and simultaneously immersing the three-electrode system into 10mL of PBS (phosphate buffer solution) solution containing 0.05mM L-tryptophan or 0.05 mMD-tryptophan and having the pH value of 7 for incubation time of 2 min. And carrying out DPV test in a potential window of 0.4-1V, and comparing current differences.
As can be seen from FIG. 2, the D-/L-tryptophan current ratio at 0.74V of the bare glassy carbon-modified electrode was 1.
As can be seen from FIG. 3, D-/L-tryptophan currents of the PEI modified electrode are substantially overlapped at 0.80V, and the current ratio is 1.
As can be seen from FIG. 4, the D-/L-tryptophan current ratio of the D-BGAc modified electrode was 1.56.
As can be seen from FIG. 5, the D-/L-tryptophan current ratio of the PEI/D-BGAc modified electrode at 0.75V is 3.4, which indicates that the recognition efficiency of the PEI/D-BGAc modified electrode on tryptophan enantiomer is remarkably improved.
Example three:
the procedure for preparing PEI/D-BGAc modified electrode is as in example two. The prepared modified electrode is used as a working electrode, an Ag/AgCl electrode is used as a reference electrode, a platinum wire electrode is used as a counter electrode, and the three-electrode system is respectively immersed into 10mL of PBS solution containing different mixing ratios of L-tryptophan and D-tryptophan. And (3) carrying out DPV test in a potential window of 0.4-1V to obtain a graph of current variation along with the content of L-tryptophan. As shown in fig. 6: the current is linear with the L-tryptophan content.
[1]Li C,Jin X,Zhao T,Zhou J,Duan P.Optically active quantum dots with induced circularly polarized luminescence in amphiphilic peptide dendron hydrogel. Nanoscale Advances 2019,1,508-512. 。
Claims (2)
1. The application of the polyethyleneimine/chiral peptide modified glassy carbon electrode in selective recognition of tryptophan enantiomer is characterized by comprising the following steps:
a. preparing a polyethyleneimine modified electrode: preparing a 1-3 mg/mL polyethyleneimine solution, and immersing the glassy carbon electrode in the polyethyleneimine solution for 20-60 min to obtain a polyethyleneimine modified electrode;
b. preparing a polyethyleneimine/chiral peptide modified electrode: preparing a chiral peptide solution, cleaning the polyethyleneimine modified electrode prepared in the step a with water, soaking the polyethyleneimine modified electrode in 0.1-1 mg/mL of chiral peptide solution for 10-30 min, taking out the polyethyleneimine modified electrode, and drying the polyethyleneimine modified electrode at room temperature to obtain the polyethyleneimine/chiral peptide modified electrode; wherein the chiral peptide is BGAc,
the BGAc structural formula is as follows:
c. electrochemical identification of tryptophan enantiomers: and (2) taking a polyethyleneimine/chiral peptide modified electrode as a working electrode, an Ag/AgCl electrode as a reference electrode and a platinum wire electrode as a counter electrode, simultaneously immersing the three-electrode system into a prepared 0.01-0.8 mM L-/D-tryptophan solution, incubating for 1-30 min, and then testing by using a pulse voltammetry method.
2. The application of the polyethyleneimine/chiral peptide modified glassy carbon electrode in selective recognition of tryptophan enantiomer is characterized in that: and c, the scanning potential in the step c is 0.4-1.0V.
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| CN109975379A (en) * | 2019-04-30 | 2019-07-05 | 常州大学 | A kind of chitosan/CT-DNA Modified Gold Electrode preparation method for electrochemical recognition Tryptophan enantiomer |
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| US7879212B2 (en) * | 2005-11-03 | 2011-02-01 | Ramot At Tel-Aviv University Ltd. | Peptide nanostructure-coated electrodes |
| JP6921217B2 (en) * | 2017-02-22 | 2021-08-18 | エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ | Polymer coating of electrodes for sensor devices |
| CN106908507B (en) * | 2017-02-22 | 2019-06-04 | 常州大学 | A kind of preparation and its application of phenylalanine dipeptide-Chitosan Composites |
| CN108645900A (en) * | 2018-04-18 | 2018-10-12 | 常州大学 | A kind of preparation method for the glutathione modified electrode can be applied to electrochemistry chiral Recognition |
| CN108562628B (en) * | 2018-04-18 | 2019-11-12 | 常州大学 | A kind of preparation for the alpha-cyclodextrin inclusion glutathione composite material modified electrode can be applied to electrochemistry chiral Recognition |
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| CN106226381A (en) * | 2016-07-11 | 2016-12-14 | 常州大学 | The preparation of a kind of Thermo-sensitive quinine chiral sensor and be applied to electrochemical recognition Tryptophan enantiomer |
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