阅读说明：本技术 一种磺化纳米丝素-纤维素混合膜及其制备方法与应用 (Sulfonated nano fibroin-cellulose mixed membrane and preparation method and application thereof ) 是由 易世雄 代方银 孙胜 童晓玲 邹雨珊 于 2020-07-03 设计创作，主要内容包括：本发明公开了一种磺化纳米丝素-纤维素混合膜及其制备方法。以丝素和醋酸纤维素为原材料,采用静电纺丝技术通过混纺制备纳米丝素-纤维素混合膜材料,采用3-羧基苯磺酸钠对纳米丝素-纤维素混合膜材料进行化学改性,制成磺化纳米丝素-纤维素混合膜。本发明首先以绿色生物质纤维丝素纤维和醋酸纤维素为原材料,使用静电纺丝技术进行混纺制备出纳米丝素-纤维素混合膜材料,接着采用3-羧基苯磺酸钠对其进行化学改性,制备出磺化纳米丝素-纤维素混合膜,其对假丝酵母脂肪酶具有较好的吸附效果,最后通过在溶液中加入中性盐将假丝酵母脂肪酶从混合膜上解吸下来进行回收,并且混合膜的重复使用性能优异。(The invention discloses a sulfonated nano fibroin-cellulose mixed membrane and a preparation method thereof. The method comprises the steps of taking fibroin and cellulose acetate as raw materials, preparing a nano fibroin-cellulose mixed membrane material by blending through an electrostatic spinning technology, and chemically modifying the nano fibroin-cellulose mixed membrane material by adopting 3-carboxyl sodium benzenesulfonate to prepare the sulfonated nano fibroin-cellulose mixed membrane. The method comprises the steps of firstly, taking green biomass fiber silk fibroin fiber and cellulose acetate as raw materials, blending by using an electrostatic spinning technology to prepare a nano silk fibroin-cellulose mixed membrane material, then, chemically modifying the nano silk fibroin-cellulose mixed membrane material by using 3-carboxyl sodium benzenesulfonate to prepare a sulfonated nano silk fibroin-cellulose mixed membrane, wherein the sulfonated nano silk fibroin-cellulose mixed membrane has a good adsorption effect on candida lipase, and finally, the candida lipase is desorbed from the mixed membrane by adding neutral salt into a solution for recycling, and the mixed membrane has excellent reusability.)

1. The sulfonated nano fibroin-cellulose mixed membrane is characterized by comprising the following components in parts by weight:

0.2-1 part of cellulose acetate;

1-3 parts of silk fibroin fibers.

2. The preparation method of the sulfonated nano silk fibroin-cellulose mixed membrane as claimed in claim 1, wherein the cellulose acetate is 0.6 part, and the silk fibroin is 1.4 parts.

3. The preparation method of the sulfonated nano silk fibroin-cellulose mixed membrane as claimed in claim 1 is characterized by comprising the following specific steps:

1) fibroin and cellulose acetate are used as raw materials, and a nano fibroin-cellulose mixed membrane material is prepared by blending by adopting an electrostatic spinning technology;

2) the nano fibroin-cellulose mixed membrane material is chemically modified by adopting 3-carboxyl sodium benzenesulfonate to prepare the sulfonated nano fibroin-cellulose mixed membrane.

4. The method for preparing the sulfonated nano silk fibroin-cellulose mixed membrane as claimed in claim 3, wherein the specific steps for preparing the nano silk fibroin-cellulose mixed membrane material in the step 1) are as follows:

1-1) carrying out degumming treatment on the silkworm cocoons at high temperature by using a sodium carbonate solution to obtain silk fibroin fibers; the concentration range of the sodium carbonate solution is 0.2-0.7 wt%, and the temperature range is 85-100 ℃;

1-2) dissolving the silk fibroin fibers in a lithium bromide solution, dialyzing and filtering, taking out the silk fibroin fibers, and freeze-drying to obtain regenerated silk fibroin; the concentration range of the lithium bromide solution is 7mol L^-1～9mol L^-1Silk fibroin fiber and bromineThe lithium dissolving solution comprises the following components in parts by weight: 1-3 parts of silk fibroin fiber and 700-900 parts of lithium bromide solution;

1-3) dissolving regenerated silk fibroin and cellulose acetate in a formic acid solution according to a certain proportion to form a spinning solution; the concentration range of the formic acid solution is 85 wt% -98 wt%, and the regenerated silk fibroin, the cellulose acetate and the formic acid solution respectively comprise the following components in parts by weight: 1-3 parts of regenerated fibroin, 0.2-1 part of cellulose acetate and 400-1000 parts of formic acid solution;

1-4) spinning the spinning solution as a raw material into a nano fibroin-cellulose mixed membrane material by adopting an electrostatic spinning technology, wherein the electrostatic spinning technology comprises the following parameters: the voltage range is 12kV to 20kV, the receiving range is 8cm to 12cm, and the perfusion speed range is 0.2mL L^-1～0.4mLL^-1The temperature range is 15-30 ℃, the humidity range is 50-70%, and the aperture range of the nozzle is 0.3-0.5 mm.

5. The method for preparing sulfonated nano silk element-cellulose mixed membrane as claimed in claim 4,

the concentration of the sodium carbonate solution in the step 1-1) is 0.5 wt%;

the concentration of the lithium bromide solution in the step 1-2) is 8mol L^-11.4 parts of silk fibroin fiber and 800 parts of lithium bromide solution;

in the step 1-3), the concentration of the formic acid solution is 98%, the weight parts of the regenerated fibroin are 2 parts, the weight parts of the cellulose acetate are 0.6 part, and the weight parts of the formic acid solution are 600 parts.

6. The method for preparing the sulfonated nano silk fibroin-cellulose mixed membrane as claimed in claim 3, wherein the specific method for preparing the sulfonated nano silk fibroin-cellulose mixed membrane in the step 2) is as follows:

2-1) preparing a 3-carboxyl sodium benzenesulfonate solution; the concentration range is (2g L)^-1～8g L^-1)；

2-2) concentration range of 2g L^-1～8g L^-1Adding the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide into the 3-sodium carboxybenzenesulfonate solution, fully stirring at room temperature for about 60-120 minutes to prepare a mixtureA solution; the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and the 3-sodium carboxybenzenesulfonate solution respectively comprise the following components in parts by weight: 1-6 parts of 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) and 2-8 parts of 3-sodium carboxybenzenesulfonate solution;

2-3) putting the nano fibroin-cellulose mixed membrane material into the mixed solution, fully stirring for about 80-120 minutes, taking out the nano fibroin-cellulose mixed membrane material after the reaction is finished, and fully washing and drying to obtain the sulfonated nano fibroin-cellulose mixed membrane; the nanometer silk fibroin-cellulose mixed membrane material and the mixed solution respectively comprise the following components in parts by weight: 1-6 parts of a nano fibroin-cellulose mixed membrane material and 600-1000 parts of a mixed solution.

7. The method for preparing sulfonated nano silk element-cellulose mixed membrane as claimed in claim 6,

the concentration of the sodium 3-carboxybenzenesulfonate solution in step 2-1) is 5g L^-1；

The concentration of 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) in step 2-2) was 5g L^-16 parts of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and 800 parts of 3-sodium carboxybenzenesulfonate solution;

in the step 2-3), the weight parts of the nano fibroin-cellulose mixed membrane material are 1-5 parts, and the weight parts of the mixed solution are 1200 parts.

8. Use of a sulfonated nano-silk fibroin-cellulose mixed membrane as defined in claim 1 or 2 for the recovery of candida lipase.

Technical Field

The invention relates to the technical field of candida lipase recovery, in particular to a sulfonated nano fibroin-cellulose mixed membrane and a preparation method and application thereof.

Background

Nearly 60 species of microorganisms are found to produce lipase, and nearly 20 species are produced industrially. The microbial lipase has wider action pH range, action temperature range and substrate specificity than animal and plant enzymes, and is convenient for industrial production and high-purity preparation obtaining. In recent years, microbial lipases have been rapidly developed in the fields of theoretical research and practical application of enzymes, and have been widely used in the industries of food, leather, grease chemical industry and the like, and in the fields of detergents, cosmetics, medicines and the like. The research on microbial lipase is carried out in China in the 60 th century of the 20 th century, and candida lipase preparations are developed and supplied to markets in 1969. Candida lipase is a semi-ascosporogenous, non-pathogenic yeast that is generally considered safe for humans and other life forms. The lipase colony is milk white, the surface is wrinkled, and the lipase colony has wide application in the fields of chemical industry, energy, medical treatment, food and the like, and is one of the most widely commercialized lipases at present. At present, the candida lipase purification technology mainly comprises methods such as a direct extraction method, an ion exchange method and an ultrafiltration method, but the methods all have corresponding defects, for example, the enzyme activity is lost, and the method cannot be repeatedly used.

The name is as follows: candida rugosa lipase immobilized Immobilization for geranyl Butyrate Synthesis, Candida rugosa lipase for the Synthesis of 6 amino butyl Butyrate [ journal paper ] royal celluloid, Von \39561WANGLu, FENG Biao- "food and Biotechnology article" 2019 journal paper of 11 th, macroporous resins of different specifications are adopted as carriers to solidify the lipase, and the defects are that: low repeated utilization rate of lipase, complex process, difficult recovery, large pollution of chemical reagents and the like.

Disclosure of Invention

The first purpose of the invention is to provide a sulfonated nano silk fibroin-cellulose mixed membrane.

The second purpose of the invention is to provide a preparation method of the sulfonated nano silk fibroin-cellulose mixed membrane.

The third purpose of the invention is to discuss the application of the sulfonated nano silk fibroin-cellulose mixed membrane in the recovery of candida lipase.

In order to achieve the purpose, the invention adopts the following technical scheme:

the sulfonated nano fibroin-cellulose mixed membrane comprises the following components in parts by weight:

0.2-1 part of cellulose acetate;

1-3 parts of silk fibroin fibers.

Further, 0.6 part of cellulose acetate and 1.4 parts of silk fibroin are adopted.

Further, the preparation method of the sulfonated nano fibroin-cellulose mixed membrane comprises the following specific steps:

1) fibroin and cellulose acetate are used as raw materials, and a nano fibroin-cellulose mixed membrane material is prepared by blending by adopting an electrostatic spinning technology;

2) the nano fibroin-cellulose mixed membrane material is chemically modified by adopting 3-carboxyl sodium benzenesulfonate to prepare the sulfonated nano fibroin-cellulose mixed membrane.

Further, the specific steps for preparing the nano fibroin-cellulose mixed membrane material in the step 1) are as follows:

1-1) carrying out degumming treatment on the silkworm cocoons at high temperature by using a sodium carbonate solution to obtain silk fibroin fibers; the concentration range of the sodium carbonate solution is 0.2-0.7 wt%, and the temperature range is 85-100 ℃;

1-2) dissolving the silk fibroin fibers in a lithium bromide solution, dialyzing and filtering, taking out the silk fibroin fibers, and freeze-drying to obtain regenerated silk fibroin; the concentration range of the lithium bromide solution is 7mol L^-1～9mol L^-1The silk fibroin fiber and the lithium bromide solution respectively comprise the following components in parts by weight: 1-3 parts of silk fibroin fiber and 700-900 parts of lithium bromide solution;

1-3) dissolving regenerated silk fibroin and cellulose acetate in a formic acid solution according to a certain proportion to form a spinning solution; the concentration range of the formic acid solution is 85 wt% -98 wt%, and the regenerated silk fibroin, the cellulose acetate and the formic acid solution respectively comprise the following components in parts by weight: 1-3 parts of regenerated fibroin, 0.2-1 part of cellulose acetate and 400-1000 parts of formic acid solution;

1-4) spinning the spinning solution as a raw material into a nano fibroin-cellulose mixed membrane material by adopting an electrostatic spinning technology, wherein the electrostatic spinning technology comprises the following parameters: the voltage range is 12kV to 20kV, the receiving range is 8cm to 12cm, and the perfusion speed range is 0.2mL L^-1～0.4mL L^-1The temperature range is 15-30 ℃, the humidity range is 50-70%, and the aperture range of the nozzle is 0.3-0.5 mm.

Further, the concentration of the sodium carbonate solution in the step 1-1) is 0.5 wt%;

the concentration of the lithium bromide solution in the step 1-2) is 8mol L^-11.4 parts of silk fibroin fiber and 800 parts of lithium bromide solution;

in the step 1-3), the concentration of the formic acid solution is 98%, the weight parts of the regenerated fibroin are 2 parts, the weight parts of the cellulose acetate are 0.6 part, and the weight parts of the formic acid solution are 600 parts.

Further, the specific method for preparing the sulfonated nano silk fibroin-cellulose mixed membrane in the step 2) is as follows:

2-1) preparing a 3-carboxyl sodium benzenesulfonate solution; the concentration range is (2g L)^-1～8g L^-1)；

2-2) concentration range of 2g L^-1～8g L^-1Adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (D) to a sodium 3-carboxybenzenesulfonate solution, charging the solution at room temperatureStirring for about 60-120 min to prepare a mixed solution; the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and the 3-sodium carboxybenzenesulfonate solution respectively comprise the following components in parts by weight: 1-6 parts of 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) and 2-8 parts of 3-sodium carboxybenzenesulfonate solution;

2-3) putting the nano fibroin-cellulose mixed membrane material into the mixed solution, fully stirring for about 80-120 minutes, taking out the nano fibroin-cellulose mixed membrane material after the reaction is finished, and fully washing and drying to obtain the sulfonated nano fibroin-cellulose mixed membrane; the nanometer silk fibroin-cellulose mixed membrane material and the mixed solution respectively comprise the following components in parts by weight: 1-6 parts of a nano fibroin-cellulose mixed membrane material and 600-1000 parts of a mixed solution.

Further, the concentration of the sodium 3-carboxybenzenesulfonate solution in step 2-1) is 5g L^-1；

The concentration of 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) in step 2-2) was 5g L ^-16 parts of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and 800 parts of 3-sodium carboxybenzenesulfonate solution;

in the step 2-3), the weight parts of the nano fibroin-cellulose mixed membrane material are 1-5 parts, and the weight parts of the mixed solution are 1200 parts.

Further, the sulfonated nano silk fibroin-cellulose mixed membrane is applied to recovery of candida lipase.

Due to the adoption of the technical scheme, the invention has the following advantages:

the invention applies the sulfonated nano fibroin-cellulose mixed membrane to the absorption and recovery of candida lipase. The invention discloses a sulfonated nano fibroin-cellulose mixed membrane, which is prepared by blending two fibers in an electrostatic spinning mode and chemically modifying the fibers by adopting 3-sodium carboxybenzenesulfonate, wherein negative ion groups on molecular chains of the sulfonated nano fibroin-cellulose mixed membrane have a strong adsorption effect on candida lipase with positive charges in an aqueous solution.

Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof.

Drawings

Detailed Description

The invention is further illustrated by the following figures and examples.