阅读说明：本技术 一种透水红外光在生物分子合成中的应用方法 (Application method of permeable infrared light in biomolecule synthesis ) 是由 江雷 宋波 张峰 李娜 于 2021-06-17 设计创作，主要内容包括：本发明公开了一种透水红外光在生物分子合成中的应用方法,包括以下步骤：S1、制备透水红外光；S2、将步骤S1中的透水红外光照射生物合成体系,进行选择性断裂或生成分子的价键；S3、设置反应程序,将步骤S1中的透水红外光的频率、强度以及照射时间进行程序化处理或微流控制调节；S4、通过生物合成体系中的微流控通道,位于不同反应位置进行程序的补加反应物；S5、重复上述进行生物合成反应的流水化。根据本发明,通过透水红外光可辅助生物合成,利用特定频率的红外光或者脉冲照射生物分子,可精准的调控价键的断裂和生成。(The invention discloses an application method of permeable infrared light in biomolecule synthesis, which comprises the following steps: s1, preparing permeable infrared light; s2, irradiating the permeable infrared light in the step S1 on the biosynthesis system to selectively break or generate the valence bonds of molecules; s3, setting a reaction program, and performing programming processing or micro-fluidic control adjustment on the frequency, intensity and irradiation time of the permeable infrared light in the step S1; s4, adding reactants in different reaction positions for programs through microfluidic channels in a biosynthesis system; s5, repeating the above steps to streamline the biosynthesis reaction. According to the invention, the biological synthesis can be assisted by the permeable infrared light, and the breakage and generation of the valence bond can be accurately regulated and controlled by irradiating the biomolecule with infrared light or pulse with specific frequency.)

1. An application method of permeable infrared light in biomolecule synthesis is characterized by comprising the following steps:

s1, preparing permeable infrared light;

s2, irradiating the permeable infrared light in the step S1 on the biosynthesis system to selectively break or generate the valence bonds of molecules;

s3, setting a reaction program, and performing programmed processing or microfluidic control on the frequency, intensity and irradiation time of the permeable infrared light in the step S1;

s4, adding reactants in different reaction positions for programs through microfluidic channels in a biosynthesis system;

s5, repeating the above steps to streamline the biosynthesis reaction.

2. The method as claimed in claim 1, wherein the step S1 comprises the following steps:

s11, preparing a common light source or a black body radiation light source;

s12, placing a water layer structure at one side of the light source in the step S11, wherein the water layer structure filters infrared light absorbed by water;

and S13, placing a light splitting device at one side of the water layer structure in the S12, wherein the light splitting device is used for selecting specific frequencies in the permeable infrared light.

3. The method as claimed in claim 2, wherein the specific frequency of the transparent infrared ray is 30-90 thz.

4. The method as claimed in claim 1, wherein the microfluidic channel in step S4 is provided with a plurality of fluid infusion inlets, the infrared light with different frequencies and the reaction system sequentially flow through different positions of the channel, and the irradiation with different time and intensity is performed by controlling the flow rate or the residence time.

5. The method as claimed in claim 1, wherein the permeable infrared light is used for correcting the misfolding of long-chain polypeptide synthesis or melting and denaturing of biomolecules.

6. The method as claimed in claim 1, wherein the permeable infrared light is used for precise covalent bond connection or chemical modification of specific chemical groups.

Technical Field

The invention relates to the technical field of biomolecule synthesis, in particular to an application method of permeable infrared light in biomolecule synthesis.

Background

Electromagnetic waves with wavelengths between 760 nanometers and 1 millimeter are called infrared light/light. In 1800 years, he heischel, a scientist in the uk, used thermometers and triple prisms to infer the presence of infrared light. Currently, infrared light is widely applied to the fields of communication, detection, military, medical treatment and the like.

As a result of previous researches, water has three main absorption peaks (regions, see figure 1) for infrared light, and two non-absorption regions, namely, regions of permeable infrared light to be protected by the invention are formed among the three absorption peaks, wherein the permeable infrared light is infrared light capable of passing through water, and is respectively 30THz-50 THz and 55THz-90 THz. Due to the absorption characteristic of water to infrared light, the invention can directly use water as an optical filter to filter the infrared light, thereby obtaining the permeable infrared light. The importance of the water-permeable infrared light in biomedicine is the main reason and purpose for reporting the patent of the invention. Because the photothermal conversion property of infrared light can enable water molecules and biological macromolecules to generate a thermal effect, the photothermal conversion efficiency of infrared light is high, the tissue penetration depth is large, and the like, so that the infrared light is applied to the medical field. In addition, since the frequency of infrared light is precisely matched with the valence bond or interaction level of a biomolecule, the use of infrared absorption spectroscopy can be a major means of molecular identification. However, few people directly drive the breaking and generation of molecular valence bonds by using infrared light, and the invention makes up for the blank field of the current infrared light application field. Such as denaturation of the driver DNA, cleavage of high-energy phosphate bonds in nucleoside triphosphate molecules, formation of phosphodiester bonds in nucleic acid molecules, formation of peptide bonds in polypeptides, protein molecules, and post-translational modifications of proteins, including (de) methylation, (de) phosphorylation, (de) protonation, etc.

At present, the synthesis of nucleic acid and polypeptide basically adopts solid phase synthesis method, i.e. one of the components (base, amino acid) of nucleic acid or polypeptide pre-modified on solid phase resin material is used as initial reactant of synthesis reaction, then the connection reaction is carried out in sequence according to base/amino acid sequence, during which the non-reactive group is protected, and every time the reaction of one component is completed, the reactant which is not connected is eluted, and after the final reaction is completed, the protecting group is deprotected, and the whole product is dissociated from resin for purification and identification. Since the error rate increases with the length of the sequence, the final yield is also limited by the length of the sequence. In addition, since the side chain groups of amino acids (e.g., mercapto group, hydroxyl group, carboxyl group, amino group, etc.) are not less active than the groups (carboxyl group, amino group) used for peptide bond formation, and erroneous linkage between groups is easily generated without protection, the purity requirement of the reagent with protecting groups is very high. Conventional chemical reactions, even certain enzymatic reactions, typically require the energy to drive the reaction through a thermal process. However, the driving energy based on thermal effects does not allow precise control of the breaking or formation of a given chemical bond. Therefore, such reactions always have a certain proportion of by-products, and the reaction efficiency is still to be improved.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide an application method of the permeable infrared light in the synthesis of the biomolecules, the permeable infrared light can assist the biosynthesis, and the infrared light or the pulse with specific frequency is used for irradiating the biomolecules to accurately regulate and control the breakage and the generation of the valence bonds. To achieve the above objects and other advantages in accordance with the present invention, there is provided a method for applying a permeable infrared light in biomolecule synthesis, comprising the steps of:

s1, preparing permeable infrared light;

s2, irradiating the permeable infrared light in the step S1 on the biosynthesis system to selectively break or generate the valence bonds of molecules;

s3, setting a reaction program, and performing programmed processing or microfluidic control on the frequency, intensity and irradiation time of the permeable infrared light in the step S1;

s4, adding reactants in different reaction positions for programs through microfluidic channels in a biosynthesis system;

s5, repeating the above steps to streamline the biosynthesis reaction.

Preferably, the step S1 includes the steps of:

s11, preparing a common light source or a black body radiation light source;

s12, placing a water layer structure at one side of the light source in the step S11, wherein the water layer structure filters infrared light absorbed by water;

and S13, placing a light splitting device at one side of the water layer structure in the S12, wherein the light splitting device is used for selecting specific frequencies in the permeable infrared light.

Preferably, the frequency of the light-transmitting infrared ray with the specific frequency is 30-90 terahertz.

Preferably, the microfluidic channel in step S4 is provided with a plurality of fluid infusion inlets, infrared light with different frequencies is arranged at different positions of the channel through which the reaction system flows in sequence, and irradiation at different times and intensities is completed by controlling the flow rate or the residence time.

Preferably, the permeable infrared light is used for correcting the synthetic misfolding of the long-chain polypeptide or melting and denaturing of the biomolecule.

Preferably, the permeable infrared light is used for the precise covalent bond connection or chemical modification of a specific chemical group.

Compared with the prior art, the invention has the beneficial effects that:

(1) in the process of carrying out mass amplification of nucleic acid by utilizing Polymerase Chain Reaction (PCR), the infrared light of 30-90THz can promote the hydrogen bond breakage between paired bases, so that the denaturation temperature of dsDNA is reduced, and the aim of synthesizing a large amount of nucleic acid at low temperature can be fulfilled. The reduction of the denaturation temperature can expand the selection range of the DNA polymerase used for PCR, thereby further reducing the cost and improving the PCR efficiency.

(2) The synthesis of polypeptide and protein can be accurately guided by permeating water and infrared light. Because the infrared absorption energy levels of the same chemical groups at different positions of the molecular structure are different, the invention can avoid the protection and deprotection reaction of the side chain groups (such as amino, carboxyl, sulfydryl and the like) in the traditional solid phase synthesis. By utilizing programmed irradiation of infrared light with different frequencies, the length and the precision of in vitro synthesis can be further increased, and even the flow line production of non-enzymatic synthesis can be realized.

(3) The common light source can generate infrared light with specific frequency through devices such as a filter, a grating and the like. The infrared light involved in the present invention is a non-water absorbing infrared band, which is approximately in the 30-90 terahertz (THz) region. Due to this permeability (non-absorption) of water, we can filter water directly as a primary filter. The optics may then be used to continue fine filter adjustment. This method is effective for any heat-generating object according to the blackbody radiation theory.

(4) The use of water-permeable infrared light can assist in biosynthesis, which includes the breaking and formation of covalent bonds. Because the covalent bond of the infrared light and the biological molecules can generate resonance absorption, the biological molecules are continuously or pulse-type irradiated by the infrared light with specific frequency, and the breakage and generation of the valence bond can be accurately regulated and controlled. For example, infrared light of-34 THz can break a high-energy phosphate bond to efficiently hydrolyze Adenosine Triphosphate (ATP) or deoxyribonucleoside triphosphate (dNTP), and a 5 'terminal phosphate group of a single-stranded (ss) DNA and a 3' terminal hydroxyl group of dNTP can be cross-linked in a "quasi-welding" manner to form a new phosphodiester bond, thereby synthesizing nucleic acid.

Drawings

FIG. 1 is a schematic view of a microfluidic channel of a biological reaction system for applying the method of permeating infrared light in biomolecule synthesis according to the present invention;

FIG. 2 is a schematic diagram of the application of the present invention in biomolecule synthesis, which is used for the synthesis of polypeptides and proteins, the correction of misfolding and the site-specific modification of bioactive groups;

FIG. 3 is a schematic diagram of the denaturation of double-stranded DNA assisted by permeating infrared light according to the method for applying permeating infrared light in biomolecule synthesis;

FIG. 4 is a molecular formula of hydrolysis of high energy phosphate bonds for a method of applying the present invention to the synthesis of biomolecules;

FIG. 5 is a molecular structure of phosphodiester bond formation according to the method of the present invention for applying permeable infrared light in biomolecule synthesis.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1-5, a method for applying permeable infrared light in biomolecule synthesis includes the following steps: s1, preparing permeable infrared light;

s2, irradiating the permeable infrared light in the step S1 on the biosynthesis system to selectively break or generate the valence bonds of molecules;

s3, setting a reaction program, and performing programmed processing or microfluidic control on the frequency, intensity and irradiation time of the permeable infrared light in the step S1;

s4, adding reactants in different reaction positions for programs through microfluidic channels in a biosynthesis system;

s5, repeating the above steps to streamline the biosynthesis reaction.

Further, the step S1 includes the following steps:

s11, preparing a common light source or a black body radiation light source;

s12, placing a water layer structure at one side of the light source in the step S11, wherein the water layer structure filters infrared light absorbed by water;

s13, placing a light splitting device on one side of the water layer structure in the S12, selecting specific frequency in the permeable infrared light by the light splitting device, then irradiating the biological synthesis reaction system, and assisting the breaking and generation of the designated chemical bond, wherein the light splitting device comprises an optical filter and a grating.

Further, the frequency of the light-transmitting infrared ray with the specific frequency is 30-90 terahertz.

Furthermore, the microfluidic channel in step S4 is provided with a plurality of fluid infusion inlets, infrared light with different frequencies is arranged at different positions of the channel through which the reaction system flows in sequence, and irradiation at different times and intensities is completed by controlling the flow rate or the residence time.

Furthermore, the permeable infrared light is used for correcting the long-chain polypeptide synthesis error folding or melting and denaturing the biological molecules.

Further, the permeable infrared light is used for connecting precise covalent bonds of specific chemical groups or chemically modifying.

Example 1

The infrared light can promote the hydrolysis of dntps (figure 4) and generate new phosphodiester bonds (figure 5).

Exemplary polypeptide synthesis: the reaction solution was irradiated with infrared light having an energy resonance frequency with a peptide bond (FIG. 2A).

Example protein folding: since long-chain polypeptides may be misfolded after synthesis, the present invention directly irradiates the wrong portion with infrared light of a specific frequency, thereby achieving the purpose of correcting the misfolding (fig. 2B).

Biological modification of exemplified proteins: by using the permeable infrared light with specific frequency, the corresponding reactive group (such as phosphate group, methyl group, ethyl group, proton, etc.) resonating with the frequency can be modified on the protein. Precise modification of specific amino acid residues can be achieved when resolution and alignment allow (FIG. 2C).

The number of devices and the scale of the processes described herein are intended to simplify the description of the invention, and applications, modifications and variations of the invention will be apparent to those skilled in the art.

While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in various fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the general concept defined by the appended claims and their equivalents.