阅读说明：本技术 一种制备肽基晶体材料的方法 (Method for preparing peptidyl crystal material ) 是由 李峻柏 薛慧敏 费进波 于 2020-08-12 设计创作，主要内容包括：本发明公开了一种制备肽基晶体材料的方法。该方法包括：将肽基凝胶转移至密闭容器内,向其中持续通入气体,得到所述肽基单晶。上述方法中,所述气体选自水蒸气、氨气、二氧化碳和硫化氢中至少一种；所述相转变可在常温常压下进行,相转变后所得晶体为高质量单晶。该方法简单易行,所得晶体材料结晶度高,晶型独特,可作为一种氨基酸或蛋白类晶体材料的通用制备方法。(The invention discloses a method for preparing a peptidyl crystal material. The method comprises the following steps: and transferring the peptidyl gel into a closed container, and continuously introducing gas into the peptidyl gel to obtain the peptidyl single crystal. In the above method, the gas is at least one selected from the group consisting of water vapor, ammonia gas, carbon dioxide and hydrogen sulfide; the phase transformation can be carried out at normal temperature and normal pressure, and the obtained crystal after the phase transformation is a high-quality single crystal. The method is simple and easy to implement, and the obtained crystal material has high crystallinity and unique crystal form and can be used as a general preparation method of amino acid or protein crystal materials.)

1. A method of preparing a peptidyl single crystal, comprising:

and transferring the peptidyl gel into a closed container, and continuously introducing gas into the peptidyl gel to obtain the peptidyl single crystal.

2. The method of claim 1, wherein: the gas is at least one of water vapor, ammonia gas, carbon dioxide and hydrogen sulfide;

in the step of introducing the gas, the pressure of the system is 0.8-1.2 standard atmospheric pressure; specifically 1.0 standard atmospheric pressure;

the temperature is 20-30 ℃; specifically, normal temperature;

the time for introducing the gas is 30-300 minutes; specifically 120 minutes.

3. The method according to any one of claims 1-2, wherein: in the peptidyl gel, the concentration of peptide molecules is 1.25 mg/mL-10 mg/mL; specifically 5 mg/mL;

the peptide is at least one of a biological peptide molecule and a derivative thereof;

specifically, the biological peptide molecule is selected from at least one of phenylalanine dipeptide, phenylalanine tripeptide and phenylalanine tetrapeptide;

the functional group in the biological peptide molecule derivative is at least one of Fmoc, Boc and Nap.

4. The method according to any one of claims 1 to 4, wherein: the peptidyl gel is prepared according to a method comprising the steps of:

dissolving peptide molecules in a solvent A, adding a solvent B or a solvent C, and standing to obtain the peptide.

5. The method of claim 4, wherein: the solvent A is at least one selected from hexafluoroisopropanol, dimethyl sulfoxide and hydroxylamine;

the solvent B is at least one selected from benzene, toluene, xylene, o-xylene, chloroform, carbon tetrachloride and styrene;

the solvent C is selected from water or a salt-containing aqueous solution; the mass percentage concentration of the saline solution is 0.1-1.0%;

the volume ratio of the solvent A to the solvent B or the solvent C is 1: (15-85); specifically, 1: 25;

the concentration of the peptide molecules in the solution consisting of the solvent A and the peptide molecules is 31.25 mg/mL-250 mg/mL; in particular to 125 mg/mL;

in the standing step, the time is 1-20 minutes; specifically 2 minutes.

6. A peptidyl single crystal prepared by the method of any one of claims 1 to 5.

Technical Field

The invention belongs to the technical field of biological materials, and particularly relates to a method for preparing a peptidyl crystal material.

Background

Supramolecular assembly based on non-covalent interactions is one of the effective strategies for the construction of ordered functional materials. The general supermolecule assembly process has intrinsic dynamic property and self-adaptability, and the precise regulation and control of the molecular arrangement of the inner part of the assembly can be realized through external condition stimulation (such as light, ultrasound, temperature, solvent, ionic strength and the like), so that the crystal material with rich structure and multiple functions is obtained.

Peptide molecules have the characteristics of simple structure, excellent assembly performance, easy regulation and control, good biocompatibility and the like, and are widely concerned by people. Currently, crystalline materials assembled from these peptide molecules have shown great potential for applications in many fields, such as photoelectric conversion, biomimetic catalysis, drug delivery, tissue engineering, and artificial photosynthesis (g.wei, z.su, n.p.reynolds, p.arosio, i.w.hamley, e.gazit, r.mezzenga, chem.soc.rev.,2017,46, 4661). However, it is generally difficult to obtain high-quality peptidyl crystal materials by using the conventional solution method, which greatly limits the practical application of the peptidyl crystal materials, and there is an urgent need to develop a simple and efficient universal new method for preparing peptidyl crystal materials.

Disclosure of Invention

An object of the present invention is to provide a method of preparing a peptidyl crystal material.

The method utilizes a universal method of gas molecules to mediate peptide molecule gel to be converted into crystals. The phase transformation method can be realized at normal temperature and normal pressure, and the obtained crystal is a single crystal with regular appearance and high crystallinity. The preparation method is simple and easy to operate, has universality and can be used for industrial scale-up production.

The present invention provides a method for preparing a peptide-based single crystal, comprising:

and transferring the peptidyl gel into a closed container, and continuously introducing gas into the peptidyl gel to obtain the peptidyl single crystal.

In the above method, the gas is at least one selected from the group consisting of water vapor, ammonia gas, carbon dioxide and hydrogen sulfide;

in the step of introducing the gas, the pressure of the system is 0.8-1.2 standard atmospheric pressure; specifically 1.0 standard atmospheric pressure;

the temperature is 20-30 ℃; specifically, normal temperature (25 ℃);

the time for introducing the gas is 30-300 minutes; specifically 120 or 180 or 120-180 or 100-280 minutes.

In the peptidyl gel, the concentration of peptide molecules is 1.25 mg/mL-10 mg/mL; specifically 5 mg/mL;

the peptide is at least one of a biological peptide molecule and a derivative thereof;

specifically, the biological peptide molecule is selected from at least one of phenylalanine dipeptide, phenylalanine tripeptide and phenylalanine tetrapeptide;

the functional group in the biological peptide molecule derivative is at least one of Fmoc, Boc and Nap.

The peptide may more specifically be a phenylalanine dipeptide.

The peptidyl gel can be prepared according to various conventional methods, and the structures of the peptidyl gels obtained by the various methods are not substantially different; specifically, the peptidyl gel may be prepared according to a method comprising the steps of:

dissolving peptide molecules in a solvent A, adding a solvent B or a solvent C, and standing to obtain the peptide.

Specifically, the solvent A is at least one selected from hexafluoroisopropanol, dimethyl sulfoxide and hydroxylamine;

the solvent B is at least one selected from benzene, toluene, xylene, o-xylene, chloroform, carbon tetrachloride and styrene;

the solvent C is selected from water or a salt-containing aqueous solution; the mass percentage concentration of the saline solution is 0.1-1.0%;

the volume ratio of the solvent A to the solvent B or the solvent C is 1: (15-85); specifically, 1: 25;

the concentration of the peptide molecules in the solution consisting of the solvent A and the peptide molecules is 31.25 mg/mL-250 mg/mL; in particular 125 or 50-150 mg/mL.

In the standing step, the time is 1-20 minutes; specifically 1-10 minutes or 2 minutes.

In the above method of preparing a peptidyl gel, the solvent a is used to dissolve the peptide molecule; selecting a solvent B to prepare the peptidyl organogel; the peptidyl hydrogel can be prepared by selecting the solvent C. The peptidyl gels as the raw materials have low crystallinity, and the specific structure is composed of ultra-long ultra-fine fibers.

The peptidyl gel can be more specifically phenylalanine dipeptide toluene gel, namely the phenylalanine dipeptide toluene gel is prepared by the following method: firstly, dissolving a peptide molecule phenylalanine dipeptide in a solvent A hexafluoroisopropanol, then adding a solvent B toluene, and standing to obtain the peptide molecule phenylalanine dipeptide.

In addition, the peptidyl single crystal prepared by the method also belongs to the protection scope of the invention. The peptidyl single crystal has high cleanliness.

The invention also provides structural information of the gel and the crystal before and after the peptidyl gel-crystal phase transition mediated by the gas molecules. The phase transition propeptide-based gel has no obvious crystal characteristics, and the resulting crystalline material after phase transition has high crystallinity.

The invention has the following advantages:

(1) the method for gas molecule mediated peptidyl gel-crystal phase transformation provided by the invention is simple and easy to implement, has low requirements on single crystal growth environment, and is easy for industrial large-scale production.

(2) The peptidyl crystal material after phase transition provided by the invention has regular appearance and high crystallinity.

Drawings

FIG. 1 is a photograph of a phenylalanine dipeptide toluol gel prepared in example 1 of the present invention.

FIG. 2 is a photograph of a water vapor-mediated phenylalanine dipeptide crystal prepared in example 2 of the present invention.

FIG. 3 is a photograph of an ammonia-mediated phenylalanine dipeptide crystal prepared in example 3 of the present invention.

FIG. 4 is a scanning electron micrograph of phenylalanine dipeptide toluol gel prepared in example 4 of the present invention.

FIG. 5 is a scanning electron micrograph of a water vapor-mediated phenylalanine dipeptide crystal prepared in example 4 of the present invention.

FIG. 6 is a scanning electron micrograph of an ammonia-mediated phenylalanine dipeptide crystal prepared in example 4 of the present invention.

FIG. 7 is a powder XRD analysis of the phenylalanine dipeptide toluol gel prepared in example 5 of the present invention.

FIG. 8 is an XRD analysis chart of the water vapor-mediated phenylalanine dipeptide crystal prepared in example 5 of the present invention.

FIG. 9 is an XRD analysis chart of ammonia-mediated phenylalanine dipeptide crystal prepared in example 5 of the present invention.

Detailed Description

The present invention is described below with reference to specific embodiments, but the present invention is not limited thereto, and any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Reagents, biomaterials, etc. used in the following examples are commercially available unless otherwise specified.

In the quantitative tests in the following examples, three replicates were set up and the results averaged.

In the examples described below, phenylalanine dipeptide is known under the English name di-L-phenylanine (Phe-Phe) and is available from Sigma under the trade name P4126.