阅读说明：本技术 一种基于硅矿化微囊固定化多酶生产肌醇的方法 (Method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme ) 是由 游淳 韩平平 于 2019-10-25 设计创作，主要内容包括：本发明公开了一种基于硅矿化微囊固定化多酶生产肌醇的方法,包括：硅矿化微囊固定化多酶以及制备得到的固定化多酶用于肌醇的生产。本发明提出的固定化多酶制备肌醇的方法过程简便,条件温和,其优点在于：微囊的空心内核不仅能够包埋大量的酶分子,而且能够赋予包埋至其中的酶分子良好的物理化学微环境,增强酶分子的稳定性；微囊的囊壁具有半渗透性,能够实现底物/产物的传递交换,有利于酶促反应的进行。特别地,固定化多酶制备肌醇,能够实现酶的回收利用。相比于纯酶的催化转化,固定化多酶的多次循环使用,大大降低多次肌醇制备所需投入的酶用量,降低生产成本。(The invention discloses a method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme, which comprises the following steps: the silicon mineralized microcapsule immobilized multienzyme and the prepared immobilized multienzyme are used for producing inositol. The method for preparing inositol by immobilized multienzyme provided by the invention has the advantages of simple process, mild conditions and the following advantages: the hollow core of the microcapsule can embed a large number of enzyme molecules, can endow the embedded enzyme molecules with good physical and chemical microenvironments, and enhances the stability of the enzyme molecules; the microcapsule wall is semi-permeable, and can realize substrate/product transfer exchange, which is favorable for enzymatic reaction. In particular, the immobilized multienzyme is used for preparing inositol, and the recycling of the enzyme can be realized. Compared with the catalytic conversion of pure enzyme, the immobilized multi-enzyme can be recycled for multiple times, so that the enzyme dosage required by multiple inositol preparation is greatly reduced, and the production cost is reduced.)

1. A method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme is characterized in that the immobilized multienzyme is obtained from the multienzyme produced by silicon mineralized microcapsule immobilized inositol, and the immobilized multienzyme is applied to inositol production.

The silicon mineralized microcapsule immobilized multienzyme is prepared by adopting a method comprising the following steps:

(1) four major enzyme molecules involved in inositol production are: pre-mixing glucan phosphorylase, glucose phosphate mutase, inositol-3-phosphate synthase and inositol monophosphate, adding the enzyme solution into a calcium chloride solution, then pouring a sodium carbonate solution with equal molar concentration and equal volume into the solution, stirring, carrying out solid-liquid separation, and collecting a solid product, namely the calcium carbonate microspheres containing the multi-enzyme.

(2) And mixing the calcium carbonate microspheres containing the multienzyme with a polyethyleneimine solution, and collecting a solid product, namely polyethyleneimine-calcium carbonate microspheres, after solid-liquid separation.

(3) Mixing the polyethyleneimine-calcium carbonate microspheres with a silicate solution, carrying out solid-liquid separation, and collecting a solid product, namely the silica mineralized-calcium carbonate microspheres.

(4) Mixing the silicon mineralized-calcium carbonate microspheres with ethylenediamine tetraacetic acid for reaction to remove calcium carbonate, and collecting a solid product, namely the silicon mineralized microcapsule immobilized multienzyme after solid-liquid separation.

2. The immobilized multienzyme according to claim 1, wherein the multienzyme mixture is a multienzyme mixture used in the process of producing inositol by an amylase method;

preferably, the multi-enzyme mixture comprises: glucanotransferase, glucose phosphorylase, inositol 3-phosphate synthase and inositol monophosphatase;

more preferably, the glucan phosphorylase is used in an amount of 0.5mg/ml, the phosphoglucomutase is used in an amount of 0.5mg/ml, the inositol-3-phosphate synthase is used in an amount of 3.0mg/ml, and the inositol monophosphatase is used in an amount of 0.4 mg/ml.

3. The immobilized multienzyme according to claim 1, wherein the immobilized multienzyme is obtained by entrapping a multienzyme mixture for inositol production in silica-mineralized microcapsules.

4. The immobilized multienzyme according to claim 1, wherein the concentration of polyethyleneimine is 0.1-1.0g/L, and the molecular weight of polyethyleneimine is 600-70000.

5. The immobilized multienzyme according to claim 1, wherein the concentration of the silicate is 2-10g/L, more preferably the silicate is sodium silicate.

6. The immobilized multienzyme according to claim 1, wherein the number of mineralized layers of the silicon mineralized microcapsules is 1-3.

7. The immobilized multienzyme according to any one of claims 1 to 5, wherein the ratio of the mass of the polyethyleneimine to the mass of the calcium carbonate microspheres comprising the multienzyme is 20-50: 1.

8. The immobilized multienzyme according to any one of claims 1 to 5, wherein the ratio of the mass of the silicate to the mass of the calcium carbonate microspheres comprising the multienzyme is 20 to 50: 1.

9. The immobilized multienzyme according to claim 1, wherein the silicon mineralized microcapsule immobilized multienzyme is prepared by a method comprising the following steps:

(1) the four major enzyme molecules involved in inositol production: the glucan phosphorylase, the phosphoglucomutase, the inositol-3-phosphate synthase and the inositol monophosphatase are premixed according to a specific enzyme dosage, wherein the dosage of the glucan phosphorylase is 0.1-1mg/ml, the dosage of the phosphoglucomutase is 0.1-1mg/ml, the dosage of the inositol-3-phosphate synthase is 0.1-5mg/ml, and the dosage of the inositol monophosphatase is 0.1-1 mg/ml. Adding the enzyme solution into a calcium chloride solution with the concentration of 0.2-0.4M, pouring a sodium carbonate solution with equal molar concentration and equal volume into the calcium chloride solution at the rotating speed of 600-1500r/min, reacting for 20-30s, performing centrifugal separation at the rotating speed of 3000r/min, washing the supernatant by deionized water after removing the supernatant until the supernatant does not contain sodium ions and chloride ions, and obtaining the calcium carbonate microspheres containing the enzyme.

(2) Preparing silicate solution with the concentration of 2-10g/L and preparing polyethyleneimine solution with the molecular weight of 600-70000, wherein the concentration of the polyethyleneimine solution is 0.1-1.0 g/L. According to the mass ratio of 20-50: 1, uniformly mixing a polyethyleneimine solution with calcium carbonate microspheres containing enzyme for 10-20 min; centrifuging at 3000r/min, removing supernatant, and washing with deionized water until the supernatant contains no polyethyleneimine; adding a silicate solution into the microspheres according to the mass ratio of 20-50: 1, and uniformly mixing for 10-20 min; centrifuging at 3000r/min, removing supernatant, and washing with deionized water until the supernatant contains no silicate ions.

(3) Preparing 0.03-0.05M EDTA solution, and adjusting pH to 5.0-6.0; according to the mass ratio of 20-50: 1, uniformly mixing an ethylene diamine tetraacetic acid solution with the obtained microspheres, shaking for 10-20min, carrying out centrifugal separation at the rotating speed of 3000r/min, and removing supernatant; repeatedly washing with EDTA for 3-4 times; washing with deionized water until the supernatant does not contain EDTA to obtain the silicon mineralized microcapsule immobilized multienzyme.

10. The process for producing inositol by using the immobilized multienzyme according to any one of claims 1 to 8, wherein the immobilized multienzyme is used to produce inositol by an enzymatic conversion method using starch or a derivative thereof as a starting material.

Preferably, the method specifically comprises: taking 10-100g/L starch or starch derivative, 80-120mM HEPES buffer solution with the pH value of 7-7.5, 10-50mM inorganic phosphate radical, 3-7mM divalent magnesium ion, 0.3-0.7mM zinc ion or manganese ion, 0.8-1.2U/ml isoamylase and 1-10mg/ml immobilized multi-enzyme reaction solution, and carrying out enzyme catalytic conversion reaction at 65-75 ℃.

11. The method for producing inositol according to claim 9, wherein after completion of the reaction, solid-liquid separation is performed, and the immobilized multienzyme is collected and recycled for the production of inositol; preferably 1-6 times, more preferably 1-4 times.

Technical Field

The invention relates to the field of inositol production, in particular to a method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme.

Background

Inositol, also known as inositol, is one of the water-soluble B-group of vitamins. Inositol is an essential substance for growth of human beings, animals and microorganisms, and is widely applied to industries such as medicines, foods, feeds and the like. At present, the global demand of inositol is about 10,000 tons every year, and the yield in China and even all over the world is far from meeting the demand. Patent CN106148425A provides a method for preparing inositol through enzyme-catalyzed transformation, which produces inositol from starch and starch derivatives by in vitro multi-enzyme catalysis. The method has the advantages of high conversion rate of raw materials, high yield of inositol, simple steps, small influence on the environment and realization of large-scale production of the inositol. However, in the method, a plurality of enzyme molecules can be obtained only by complicated extraction and purification, and the cost is high; and the water-soluble enzyme molecules are difficult to recover after the catalysis is finished, thereby causing waste. Immobilizing enzymes and using the immobilized enzymes for catalytic reactions have many advantages: the separation and extraction of the product are simplified, and the product quality is improved; the stability of the enzyme is improved, the enzyme can be used for a long time and continuously reacts, the enzyme is recycled for a plurality of times, and the production cost is greatly reduced.

In recent years, microcapsules have attracted more and more attention as a new hollow material, and are widely applied to the fields of catalysis, sensing, drug release and the like. In particular, since the microcapsule has a structure similar to a cell, it is widely used in the field of immobilized enzymes. Enzyme molecules can be embedded in the hollow core of the microcapsule, and the semipermeable capsule wall can realize the exchange transmission of substrates/products. By regulating and controlling the composition and structure of the capsule wall, a suitable microenvironment can be provided for enzyme molecules, and the stability of the enzyme is improved.

Disclosure of Invention

The invention aims to provide a method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme, which adopts the silicon mineralized microcapsule as an immobilized enzyme carrier, immobilizes the multienzyme catalytically converted by an inositol enzyme method to obtain the immobilized multienzyme, and produces the inositol by utilizing the immobilized multienzyme catalysis, thereby realizing the recycling of the enzyme. The immobilized multienzyme can be recycled for many times, so that the enzyme dosage required by inositol preparation is greatly reduced, and the production cost is reduced.

Specifically, the invention provides a method for producing inositol based on silicon mineralized microcapsule immobilized multienzyme, which is formed by silicon mineralized microcapsule immobilized multienzyme.

The invention adopts silicon mineralized microcapsule to prepare inositol with four main enzyme molecules: glucan phosphorylase (aGP), Phosphoglucomutase (PGM), inositol-3-phosphate synthase (IPS) and Inositol Monophosphatase (IMP) were co-immobilized. The hollow core of the microcapsule can endow the enzyme molecules embedded therein with good physical and chemical microenvironments, and enhance the stability of the enzyme molecules; the microcapsule wall is semi-permeable, and can realize substrate/product transfer exchange, which is favorable for enzymatic reaction.

The silicon mineralized microcapsule immobilized multienzyme can be prepared by adopting a method comprising the following steps:

(1) four major enzyme molecules involved in inositol production are: pre-mixing glucan phosphorylase, glucose phosphate mutase, inositol-3-phosphate synthase and inositol monophosphate, adding the enzyme solution into a calcium chloride solution, then pouring a sodium carbonate solution with equal molar concentration and equal volume into the solution, stirring, carrying out solid-liquid separation, and collecting a solid product, namely the calcium carbonate microspheres containing the multi-enzyme.

(2) And mixing the calcium carbonate microspheres containing the multienzyme with a polyethyleneimine solution, and collecting a solid product, namely polyethyleneimine-calcium carbonate microspheres, after solid-liquid separation.

(3) Mixing the polyethyleneimine-calcium carbonate microspheres with a silicate solution, carrying out solid-liquid separation, and collecting a solid product, namely the silica mineralized-calcium carbonate microspheres.

(4) Mixing the silicon mineralized-calcium carbonate microspheres with ethylenediamine tetraacetic acid for reaction to remove calcium carbonate, and collecting a solid product, namely the silicon mineralized microcapsule immobilized multienzyme after solid-liquid separation.

The invention optimizes the method for preparing the silicon mineralized microcapsule immobilized multienzyme, thereby improving the structure of the silicon mineralized microcapsule and further improving the catalytic and recycling performances of the immobilized multienzyme.

In the step (2) of preparing the silica mineralized microcapsule immobilized multienzyme, the molecular weight and the concentration of polyethyleneimine determine the capsule wall thickness of the silica mineralized microcapsule. The molecular weight of the polyethyleneimine is preferably 600-70000Da, and the concentration is 0.1-1.0 g/L.

Silicate is introduced in the step (3) of preparing the silicon mineralized microcapsule immobilized multienzyme, so that inorganic mineral substances can be provided for mineralization, and the complete progress of silicon mineralization is promoted. The practice of the invention shows that when the concentration of silicate is 2-10g/L, and the silicate is preferably sodium silicate, the prepared silicon mineralized microcapsule has good catalytic activity, and particularly can improve the catalytic effect of an immobilized multi-enzyme system.

As a preferable scheme of the invention, the silicon mineralized microcapsule immobilized multienzyme is prepared by adopting a method comprising the following steps:

(1) the four major enzyme molecules involved in inositol production: mixing glucan phosphorylase, phosphoglucomutase, inositol-3-phosphate synthase and inositol monophosphatase in advance according to specific enzyme dosage, wherein the glucan phosphorylase dosage is 0.1-1mg/ml, and the phosphoglucomutase dosage is 0.1-1mg/ml

mg/ml, inositol-3-phosphate synthetase dosage is 0.1-5mg/ml, and inositol-monophosphatase dosage is 0.1-1 mg/ml. Adding the enzyme solution into a calcium chloride solution with the concentration of 0.2-0.4M, pouring a sodium carbonate solution with equal molar concentration and equal volume into the calcium chloride solution at the rotating speed of 600-1500r/min, reacting for 20-30s, performing centrifugal separation at the rotating speed of 3000r/min, washing the supernatant by deionized water after removing the supernatant until the supernatant does not contain sodium ions and chloride ions, and obtaining the calcium carbonate microspheres containing the enzyme.

(2) Preparing silicate solution with the concentration of 2-10g/L and preparing polyethyleneimine solution with the molecular weight of 600-70000, wherein the concentration of the polyethyleneimine solution is 0.1-1.0 g/L. According to the mass ratio of 20-50: 1, uniformly mixing a polyethyleneimine solution with calcium carbonate microspheres containing enzyme for 10-20 min; centrifuging at 3000r/min, removing supernatant, and washing with deionized water until the supernatant contains no polyethyleneimine; adding a silicate solution into the microspheres according to the mass ratio of 20-50: 1, and uniformly mixing for 10-20 min; centrifuging at 3000r/min, removing supernatant, and washing with deionized water until the supernatant contains no silicate ions.

(3) Preparing 0.03-0.05M EDTA solution, and adjusting pH to 5.0-6.0; according to the mass ratio of 20-50: 1, uniformly mixing an ethylene diamine tetraacetic acid solution with the obtained microspheres, shaking for 10-20min, carrying out centrifugal separation at the rotating speed of 3000r/min, and removing supernatant; repeatedly washing with EDTA for 3-4 times; washing with deionized water until the supernatant does not contain EDTA to obtain the silicon mineralized microcapsule immobilized multienzyme.

The multienzyme for inositol production according to the present invention includes glucan phosphorylase, phosphoglucomutase, inositol-3-phosphate synthase and inositol monophosphatase. When the four enzymes are jointly immobilized in the silicon mineralized microcapsule, the enzymes can be repeatedly utilized on the basis of ensuring the catalytic efficiency of the enzymes, so that the production cost is obviously reduced. The optimized enzyme proportion of the invention is that the dosage of the glucan phosphorylase is 0.1-1mg/ml, the dosage of the glucose phosphoglucomutase is 0.1-1mg/ml, the dosage of the inositol-3-phosphate synthase is 0.1-5mg/ml and the dosage of the inositol monophosphatase is 0.1-1mg/ml, thereby ensuring that when the inositol is actually produced, the immobilized multienzyme can realize good catalytic efficiency and can be recycled for a plurality of times under the condition of reasonable dosage.

The invention further protects a method for producing inositol by using the immobilized multienzyme, which takes starch or derivatives thereof as raw materials, and prepares the inositol by an enzyme catalytic conversion method by using the immobilized multienzyme.

As a preferred scheme of the present invention, the method specifically comprises: taking 10-100g/L starch or starch derivative, 80-120mM HEPES buffer solution with the pH value of 7-7.5, 10-50mM inorganic phosphate radical, 3-7mM divalent magnesium ion, 0.3-0.7mM zinc ion or manganese ion, 0.8-1.2U/ml isoamylase and 1-10mg/ml immobilized multi-enzyme reaction solution, and carrying out enzyme catalytic conversion reaction at 65-75 ℃.

After the reaction is completed, solid-liquid separation is carried out, and the immobilized multienzyme is collected and can be recycled for the preparation of inositol. The immobilized multienzyme can keep higher catalytic activity when being recycled for 1-6 times, preferably for 1-4 times.

Compared with the prior art, the method for producing inositol by silicon mineralized microcapsule immobilized multienzyme has the advantages of simple process and mild conditions. In particular, the recovery and utilization of the enzyme can be realized by preparing inositol by immobilizing a multienzyme. The enzyme can be recycled for many times, so that the enzyme dosage required by many times of inositol preparation is greatly reduced, and the production cost is reduced.

Drawings

FIG. 1 shows the transformation curve and TEM image of the immobilized multi-enzyme with 1 silicon mineralized layer for preparing inositol, provided in example 1.

FIG. 2 shows the transformation curve and TEM image of the immobilized multi-enzyme inositol prepared with 2 Si mineralization layers provided in example 2.

FIG. 3 shows the transformation curve and TEM image of the immobilized multi-enzyme with 2 silicon mineralized layers for preparing inositol, provided in example 3.

FIG. 4 shows the conversion curve for the preparation of myo-inositol from immobilized multi-enzyme with polyethyleneimine molecular weight of 600, with the number of silica mineralized layers provided in example 6 being 2.

FIG. 5 is a bar graph showing the cycling of immobilized polyases with polyethyleneimine molecular weight of 600 to produce inositol, with the number of silicon mineralized layers being 2 as provided in example 7.

Detailed Description

The invention discloses a method for producing inositol by immobilized multienzyme, which can be realized by appropriately improving process parameters by one skilled in the art with reference to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods of the present invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention.

Example 1

The embodiment provides a silicon mineralized microcapsule immobilized multienzyme, which is prepared by the following method:

(1) the four major enzyme molecules involved in the preparation of inositol in patent CN 106148425A: glucan phosphorylase, phosphoglucomutase, inositol-3-phosphate synthase and inositol monophosphate are premixed in a specific enzyme amount, wherein the glucan phosphorylase amount is 0.5mg/ml, the phosphoglucomutase amount is 0.5mg/ml, the inositol-3-phosphate synthase amount is 3.0mg/ml, and the inositol monophosphatase amount is 0.4 mg/ml. Adding the enzyme solution into a calcium chloride solution with the concentration of 0.33M, pouring a sodium carbonate solution with the equal molar concentration and the same volume into the calcium chloride solution at the rotating speed of 700r/min, reacting for 20-30s, carrying out centrifugal separation at the rotating speed of 3000r/min, removing supernatant, washing with deionized water until the supernatant does not contain sodium ions and chloride ions, and obtaining the calcium carbonate microspheres containing the enzyme.

(2) Sodium silicate solution with the concentration of 8.5g/L and polyethyleneimine solution with the molecular weight of 1800 are prepared, and the concentration of the polyethyleneimine solution is 0.5 g/L. According to the mass ratio of 30:1, uniformly mixing a polyethyleneimine solution with calcium carbonate microspheres containing enzyme for 10-20 min; centrifuging at 3000r/min, removing supernatant, and washing with deionized water until the supernatant contains no polyethyleneimine; adding a sodium silicate solution into the microspheres according to the mass ratio of 30:1, and uniformly mixing for 10-20 min; centrifuging at 3000r/min, removing supernatant, and washing with deionized water until the supernatant contains no silicate ions.

(3) Preparing an ethylenediamine tetraacetic acid solution with the concentration of 0.05M, and adjusting the pH value to 5.8; according to the mass ratio of 30:1, uniformly mixing an ethylene diamine tetraacetic acid solution with the obtained microspheres, shaking for 10-20min, carrying out centrifugal separation at the rotating speed of 3000r/min, and removing supernatant; repeatedly washing with EDTA for 3-4 times; washing with deionized water until the supernatant does not contain EDTA to obtain the silicon mineralized microcapsule immobilized multienzyme.

Example 2

This example provides a silica mineralized microcapsule immobilized multienzyme, which differs from example 1 only in that: the number of the capsule wall layers of the silicon mineralized microcapsule is changed from 1 layer to 2 layers.

Example 3

This example provides a silica mineralized microcapsule immobilized multienzyme, which differs from example 1 only in that: the number of the capsule wall layers of the silicon mineralized microcapsule is changed from 1 layer to 3 layers.

Example 4

This example provides a silica mineralized microcapsule immobilized multienzyme, which differs from example 1 only in that: the molecular weight of the polyethyleneimine is changed from 1800 to 600.

Example 5

This example provides a silica mineralized microcapsule immobilized multienzyme, which differs from example 1 only in that: the molecular weight of the polyethyleneimine was changed from 1800 to 70000.

Example 6

This example provides a silica mineralized microcapsule immobilized multienzyme, which differs from example 1 only in that: the molecular weight of the polyethyleneimine is changed from 1800 to 600; the number of the capsule wall layers of the silicon mineralized microcapsule is changed from 1 layer to 2 layers.

Comparative example

This comparative example provides an unimmobilized multienzyme mixture for the production of myo-inositol, which has the same composition as the multienzyme mixture in example 1.

Example 7

Inositol was prepared by using the immobilized multi-enzymes provided in examples 1 to 6, or the multi-enzyme mixture provided in comparative example, respectively, as follows:

taking 50g/L starch or starch derivative, 100mM HEPES buffer solution with the pH value of 7.2, 20mM inorganic phosphate radical, 5mM divalent magnesium ion, 0.5mM zinc ion or manganese ion, 1U/ml isoamylase and 5mg/ml immobilized multi-enzyme (or multi-enzyme mixture) provided by the above examples or comparative examples, carrying out enzyme catalytic conversion reaction at 70 ℃, and detecting the concentration of inositol by high performance liquid chromatography.

The immobilized multi-enzyme provided in example 1 was used for preparation, and the reaction was carried out for 37 hours to approach the reaction equilibrium, and the inositol generation concentration was 18.5g/L and the conversion was 37%.

The immobilized multi-enzyme provided in example 2 was used for preparation, the reaction was near the equilibrium in 37 hours, the inositol generation concentration was 28g/L, and the conversion was 54%.

The immobilized multienzyme preparation provided in example 3 was carried out, and after 37 hours of reaction, the equilibrium of the reaction was approached, the inositol generation concentration was 3g/L, and the conversion was 6%.

The immobilized multienzyme preparation provided in example 4 was carried out at a inositol generation concentration of 35g/L and a conversion of 70% after 21 hours of reaction and approaching the reaction equilibrium.

The immobilized multienzyme preparation provided in example 5 was carried out, and after 21 hours of reaction, the equilibrium of the reaction was approached, the inositol generation concentration was 17.2g/L, and the conversion was 34.4%.

The immobilized multi-enzyme preparation provided in example 6 was carried out, and after 20 hours of reaction, the reaction equilibrium was approached, the inositol generation concentration was 37g/L, and the conversion rate was 74%.

The preparation was carried out using the multienzyme mixture provided in the comparative example, and after 10 hours of reaction, the equilibrium of the reaction was approached, the inositol generation concentration was 38g/L, and the conversion was 76%.

Example 8

Inositol was produced by the method provided in example 7, followed by solid-liquid separation, and the immobilized multi-enzyme was collected and recycled for the production of inositol. The concentration of inositol produced during each cycle was measured by high performance liquid chromatography, and the results were expressed as relative inositol production concentration, with the concentration of inositol produced in the first cycle reaction set at 100%.

The immobilized multi-enzyme prepared by the method of example 6 maintained a relative inositol production concentration of 34% after 4 cycles of use, and the results are shown in FIG. 5.

Example 9

After inositol is prepared by the method provided by the comparative example, ultrafiltration is carried out, and the multienzyme is recovered and recycled for preparing inositol. After 2 cycles, the ultrafiltration recovery multienzyme maintained a relative inositol production concentration of 20%.

By comparing the recycling effects of example 8 and example 9, it was found that inositol can be produced from immobilized multi-enzymes and that recycling of the enzymes can be achieved. Compared with the catalytic conversion of pure enzyme, the immobilized multi-enzyme can be recycled for multiple times, so that the enzyme dosage required by multiple inositol preparation is greatly reduced, and the production cost is reduced.

Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.