CN-119736278-B - Biological enzyme for catalyzing and slowly releasing biological energy substances and fixing method thereof

Abstract

The invention relates to a biological enzyme for catalyzing and slowly releasing biological energy substances and a fixing method thereof. The biological enzyme for catalyzing and slowly releasing the biological energy substance comprises a ceramic matrix, dopamine loaded on the ceramic matrix and amyloglucosidase loaded on the dopamine, wherein the mass ratio of the ceramic matrix is 80-90%, the mass ratio of the dopamine is 5-10% and the mass ratio of the amyloglucosidase is 1-10% based on 100% of the total mass of the biological enzyme for catalyzing and slowly releasing the biological energy substance.

Inventors

• ZHU YUFANG
• ZHAO CHAOQIAN

Assignees

• 中国科学院上海硅酸盐研究所

Dates

Publication Date

20260512

Application Date

20241219

Claims (6)

1. 1. The biological enzyme for catalyzing and slowly releasing the biological energy substance is characterized by comprising a ceramic matrix, dopamine hydrochloride loaded on the ceramic matrix and amyloglucosidase loaded on the dopamine hydrochloride, wherein the particle size of the ceramic matrix is 300nm-10 mu m, and the specific surface area of the ceramic matrix is 10-50 m 2 /g; The mass ratio of the ceramic matrix is 80-90%, the mass ratio of the dopamine hydrochloride is 5-10% and the mass ratio of the amyloglucosidase is 1-10% based on 100% of the total mass of the biological enzyme for catalyzing and slowly releasing the biological energy substance.
2. 2. A method of immobilization of a biological enzyme for catalytic slow release of a biological energy substance according to claim 1, said immobilization method comprising the steps of: (1) Mixing ceramic powder with dopamine hydrochloride to obtain raw material mixed powder, adding deionized water into the mixed powder, and stirring to obtain dopamine hydrochloride-loaded ceramic powder; (2) Mixing the ceramic powder loaded with dopamine hydrochloride with amyloglucosidase, adding deionized water and carrying out shaking mixing to enable the amyloglucosidase to be fully loaded and combined on the dopamine hydrochloride, and obtaining the immobilized biological enzyme for catalyzing and slowly releasing the biological energy substances.
3. 3. The method according to claim 2, wherein in the step (1), the particle diameter of the ceramic powder is 100nm to 10 μm.
4. 4. The method according to claim 2, wherein in the step (1), the ratio of the raw material mixed powder to deionized water is 1g:5-10 mL.
5. 5. The method according to claim 2, wherein in the step (2), the usage ratio of the dopamine hydrochloride-loaded ceramic powder to deionized water is 1 g/5-50 mL.
6. 6. The method according to claim 2, wherein in the step (2), the usage ratio of the dopamine hydrochloride-loaded ceramic powder to deionized water is 1g:20-50mL.

Description

Biological enzyme for catalyzing and slowly releasing biological energy substances and fixing method thereof Technical Field The invention belongs to the technical field of tissue engineering and bone repair, and particularly relates to a biological enzyme for catalyzing and slowly releasing biological energy substances and a fixing method thereof. Background Bone defects caused by infection, trauma and various diseases (such as osteonecrosis, osteoporosis, bone tumor, etc.) constitute a serious threat to the life health and quality of life of patients. The traditional bone repair material promotes cell differentiation and activates related cell pathways mainly by releasing Ca 2+、PO43-、Mg2+ and Zn 2+ plasmas, thereby promoting the repair process of bone tissues. However, the bone defect area is often accompanied by serious damage to blood supply, which results in a blockage of the delivery channel of bioenergy substances (including glucose, fatty acids, amino acids, etc.), thereby causing the tissue and cells at the bone defect area to lack necessary nutritional support, and finally affecting the effect of bone repair. Cells acquire Adenosine Triphosphate (ATP) by decomposing glucose, fatty acids and amino acids obtained from food or stored in the body to meet the normal energy requirements of the cells, with glucose being the energy substance of choice for most cells in the body. Then, the direct release of glucose at the defect is the best option to address the lack of nutrients at the bone defect, especially at the early stage. Biological enzymes are organic substances produced by living cells and having a catalytic function, and can significantly reduce the activation energy of chemical reactions, thereby accelerating the rate of chemical reactions. Depending on the type of substrate and catalytic reaction, biological enzymes can be further subdivided into various types, if gums, lipases, amylases, cellulases, etc. The biological enzyme has the advantages of high catalytic activity, fast catalytic efficiency, low consumption, mild reaction condition and the like, and has wide application in the fields of pharmacy, cosmetics, paper pulp, biological fuel, textile, food, water treatment, petroleum, brewing, biological medical treatment and the like. Glucose, which is required by the human body, is usually present in the food in the form of carbohydrates, such as starch. Carbohydrate-rich foods, such as rice, pasta, bread, fruits and vegetables, are important sources of glucose. When ingested by humans, the digestive system breaks down these foods into glucose monosaccharides by means of various biological enzymes (salivary amylase, pancreatic amylase and intestinal starch) and then absorbs them into human tissues to provide energy. Therefore, the in-situ catalytic slow release of glucose can be realized by utilizing biological enzymes to solve the problem of lack of energy substances at the bone defect. It was found that amyloglucosidase can effect catalytic production of glucose monosaccharides from starch. However, the application of the enzyme has the problem that the activity of the biological enzyme is easily interfered by various factors, such as temperature, various salts in the solution, concentration and pH value changes, and the like, and the factors can cause the structure of the biological enzyme to be changed, so that the catalytic activity is lost. In addition, it is also susceptible to deactivation and loss of catalytic activity over time due to its structural instability. Therefore, how to improve the stability of biological enzymes is important. Disclosure of Invention Aiming at the technical problem of poor stability of amyloglucosidase, the invention aims to provide a fixing method for improving the stability of amyloglucosidase. In a first aspect, the present invention provides a bioenzyme for catalyzing and releasing a bioelectric energy substance, the bioenzyme for catalyzing and releasing a bioelectric energy substance comprising a ceramic matrix, dopamine supported on the ceramic matrix, and amyloglucosidase supported on the dopamine; the mass ratio of the ceramic matrix is 80-90%, the mass ratio of the dopamine is 5-10% and the mass ratio of the amyloglucosidase is 1-10% based on 100% of the total mass of the biological enzyme for catalyzing and slowly releasing the biological energy substance. Preferably, the ceramic matrix is at least one of hydroxyapatite, beta-TCP and dicalcium silicate, and preferably beta-TCP. In a second aspect, the present invention provides a method for immobilizing a biological enzyme for catalytic slow release of a biological energy substance as described above, the immobilization method comprising the steps of: (1) Mixing ceramic powder with dopamine hydrochloride to obtain raw material mixed powder, adding deionized water into the mixed powder, and stirring to obtain dopamine-loaded ceramic powder; (2) Mixing the ceramic powder loaded with dopamine wit