阅读说明：本技术 一种基于大豆深加工废液以asp干粉精制sbti工艺 (Process for refining SBTI from soybean deep processing waste liquid by ASP dry powder ) 是由 梁晓晖 杨俊朋 柏冠军 曹居祥 方长林 于 2019-08-27 设计创作，主要内容包括：本发明公开了一种基于大豆深加工废液以ASP干粉精制SBTI工艺,本发明具体包括以下步骤：S1、原料的选取和称量,S2、原料的溶解,S3、溶液的脱盐处理,S4、溶液的层析和纯化处理,S5、超滤膜浓缩和混合处理,S6、SBTI成品的制备,本发明涉及生物工程技术领域。该基于大豆深加工废液以ASP干粉精制SBTI工艺,可实现通过从大豆清废水中提取的ASP干粉,来直接进行SBTI精制处理,很好的达到了通过大豆深加工废液的回收利用,来快速提取SBTI的目的,实现了节省SBTI的精制成本和提高SBTI的生产效率,同时实现了通过直接对ASP干粉进行精制SBTI,来提高SBTI精制的纯度,大大提高了生产效率,且节省了生产成本,从而对SBTI的精制十分有益。(The invention discloses a process for refining SBTI from soybean deep processing waste liquid by ASP dry powder, which specifically comprises the following steps: s1, selecting and weighing raw materials, S2, dissolving the raw materials, S3, desalting the solution, S4, carrying out chromatography and purification treatment on the solution, S5, concentrating and mixing an ultrafiltration membrane, and S6, and preparing an SBTI finished product, and relates to the technical field of bioengineering. This refined SBTI technology with ASP dry powder based on soybean deep-processing waste liquid can realize through the ASP dry powder that draws from the clear waste water of soybean, come direct SBTI refining treatment, fine reaching through the recycle of soybean deep-processing waste liquid, come the purpose of quick extraction SBTI, realized saving SBTI's refining cost and improving SBTI's production efficiency, realized simultaneously through directly refining SBTI to the ASP dry powder, improve SBTI refined purity, greatly improved production efficiency, and saved manufacturing cost, thereby it is very useful to SBTI's refining.)

1. A process for refining SBTI by ASP dry powder based on soybean deep processing waste liquid is characterized in that: the method specifically comprises the following steps:

s1, selecting and weighing raw materials: firstly, weighing ASP dry powder with required weight by weighing equipment, and transferring the weighed ASP dry powder into a storage tank for storage for use;

s2, dissolving raw materials: pouring the ASP dry powder weighed in the step S1 into a dissolving bottle, then adding water with a corresponding volume into the dissolving bottle, and stirring for 30-40min by a stirring rod to completely dissolve the ASP dry powder;

s3, desalting solution: transferring the solution dissolved in the step S2 into desalting equipment, filtering macromolecular dissolved salt through an RO reverse osmosis desalination membrane by using mechanical pressure, and allowing the filtered solution to flow into a collection tank for centralized collection treatment so as to complete desalting treatment;

s4, and carrying out chromatography and purification treatment on the solution: transferring the desalted solution obtained in the step S3 to a purification device, performing chromatography on the solution by DEAE ion exchange chromatography, and then purifying and collecting BBI protein and KTI protein;

s5, ultrafiltration membrane concentration and mixing treatment: adjusting the pH value of the solution purified in the step S4 to 5-6, concentrating by using an ultrafiltration membrane, and adding the macromolecular substances subjected to the ultrafiltration membrane and the BBI protein and the KTI protein purified in the step S4 into a mixing device for mixing treatment;

s6, preparation of SBTI finished products: and S5, transferring the mixed solution into a filtering device after the mixing is finished, filtering the mixed solution through a filter membrane, drying the filtered substance at the drying temperature of 45-55 ℃, and drying to obtain the SBTI finished product.

2. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: the weight of the ASP dry powder measured in the step S1 is 3.4-6.7 g.

3. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: in the step S2, the volume of water added into the dissolution bottle is 35-55 mL.

4. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: in the step S3, the mechanical pressure is controlled to be 0.4-0.6 Mpa.

5. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: the ion exchanger used in the DEAE ion exchange chromatography in the step S4 is one or a combination of two of carboxymethyl cellulose or diethylaminoethyl cellulose.

6. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: the molecular weight of the ultrafiltration membrane in the step S5 is 3 kDa.

7. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: when the mixing is performed in the mixing device in the step S5, the rotation speed of the stirring blade in the mixing device is 200-300r/min, and the stirring is performed for 30-40 min.

8. The process of claim 1, wherein the SBTI is refined from soybean deep processing waste liquid by ASP dry powder, and the process comprises the following steps: the drying mode in the step S6 is one of freeze drying or low-temperature spray drying.

Technical Field

The invention relates to the technical field of bioengineering, in particular to a process for refining SBTI by ASP dry powder based on soybean deep processing waste liquid.

Background

Soybean Trypsin inhibitor (SBTI) is protein pharmaceutic adjuvant obtained by naturally extracting and refining soybeans. The SBTI mainly comprises two proteins, namely a BBI (Bowman-Birk inhibitor with the molecular weight of about 8kDa) and a KTI (Kunitz trypsin inhibitor with the molecular weight of about 21kDa), wherein the BBI can inhibit the activity of trypsin, the KTI can inhibit the activities of trypsin and chymotrypsin, ASP dry powder is crude SBTI protein, the BBI protein and the KTI protein are formed according to a certain proportion in an SBTI product, in the development of a new soft capsule medicine for oral administration of polypeptides or proteins, the SBTI is used as a protective agent of active medicine components, the active components are prevented from being degraded by various digestive enzymes in a human body when passing through a gastrointestinal tract, and the new oral administration medicine of the polypeptides or proteins has very wide market prospect, so the SBTI has very important commercial application value.

The soybean clear water waste liquid is commonly used in non-transgenic soybean deep processing food enterprises by adopting an alkali dissolution and acid precipitation process to produce soybean protein isolate and edible soybean fiber by taking the soybean meal without oil as a raw material, in the acid precipitation step (pH is about 4.5), after the soybean protein isolate is precipitated, a horizontal decanter centrifuge is used for separation to obtain the soybean protein isolate and 'clear bean water', the clear bean water is generally used as waste liquid to be directly treated by a decontamination water treatment system, the clear bean water not only has large volume, but also has high BOD and COD, the wastewater treatment cost is higher, the soybean clear water usually contains 0.5 to 1 percent of soybean protein, 1 to 1.5 percent of soybean oligosaccharide, 0.5 percent of inorganic salt and the like, meanwhile, the soy albumin also contains a small amount of pigment, phytic acid, jelly and the like, and the soy albumin contains a soybean trypsin inhibitor and can be extracted by a proper process, so that waste is changed into valuable.

The prior SBTI refining process mostly adopts extraction raw materials to carry out full-flow extraction and refining, however, the SBTI obtained by the refining method has lower purity, low production efficiency and higher cost, ASP dry powder extracted from soybean clear wastewater cannot be used for directly carrying out SBTI refining treatment, the aim of rapidly extracting the SBTI by recycling soybean deep processing waste liquid cannot be achieved, the refining cost of the SBTI cannot be saved, the production efficiency of the SBTI cannot be improved, the SBTI cannot be directly refined by the ASP dry powder, the purity of the SBTI is improved, and great inconvenience is brought to the refining of the SBTI.

Disclosure of Invention

Technical problem to be solved

Aiming at the defects of the prior art, the invention provides a process for refining SBTI by ASP dry powder based on soybean deep processing waste liquid, which solves the problems that the SBTI obtained by the existing refining method has low purity, low production efficiency and high cost, the SBTI refining treatment can not be directly carried out by the ASP dry powder extracted from soybean clear wastewater, the SBTI can not be rapidly extracted by recycling the soybean deep processing waste liquid, the SBTI refining cost can not be saved and the SBTI production efficiency can not be improved, and the SBTI refining purity can not be improved by directly refining the ASP dry powder.

(II) technical scheme

In order to achieve the purpose, the invention is realized by the following technical scheme: a process for refining SBTI by ASP dry powder based on soybean deep processing waste liquid comprises the following steps:

s1, selecting and weighing raw materials: firstly, weighing ASP dry powder with required weight by weighing equipment, and transferring the weighed ASP dry powder into a storage tank for storage for use;

s2, dissolving raw materials: pouring the ASP dry powder weighed in the step S1 into a dissolving bottle, then adding water with a corresponding volume into the dissolving bottle, and stirring for 30-40min by a stirring rod to completely dissolve the ASP dry powder;

s3, desalting solution: transferring the solution dissolved in the step S2 into desalting equipment, filtering macromolecular dissolved salt through an RO reverse osmosis desalination membrane by using mechanical pressure, and allowing the filtered solution to flow into a collection tank for centralized collection treatment so as to complete desalting treatment;

s4, and carrying out chromatography and purification treatment on the solution: transferring the desalted solution obtained in the step S3 to a purification device, performing chromatography on the solution by DEAE ion exchange chromatography, and then purifying and collecting BBI protein and KTI protein;

s5, ultrafiltration membrane concentration and mixing treatment: adjusting the pH value of the solution purified in the step S4 to 5-6, concentrating by using an ultrafiltration membrane, and adding the macromolecular substances subjected to the ultrafiltration membrane and the BBI protein and the KTI protein purified in the step S4 into a mixing device for mixing treatment;

s6, preparation of SBTI finished products: and S5, transferring the mixed solution into a filtering device after the mixing is finished, filtering the mixed solution through a filter membrane, drying the filtered substance at the drying temperature of 45-55 ℃, and drying to obtain the SBTI finished product.

Preferably, the weight of the ASP dry powder measured in the step S1 is 3.4 to 6.7 g.

Preferably, the volume of water added to the dissolution flask in the step S2 is 35 to 55 mL.

Preferably, the mechanical pressure in step S3 is controlled at 0.4-0.6 MPa.

Preferably, the ion exchanger used in DEAE ion exchange chromatography in step S4 is one or a combination of two of carboxymethyl cellulose and diethylaminoethyl cellulose.

Preferably, the molecular weight filtered by the ultrafiltration membrane in the step S5 is 3 kDa.

Preferably, when the mixing is performed in the mixing device in the step S5, the rotation speed of the stirring blade in the mixing device is 200-300r/min, and the stirring is performed for 30-40 min.

Preferably, the drying manner in step S6 is one of freeze drying or low-temperature spray drying.

(III) advantageous effects

The invention provides a process for refining SBTI from soybean deep processing waste liquid by ASP dry powder. Compared with the prior art, the method has the following beneficial effects: the process for refining the SBTI by the ASP dry powder based on the soybean deep processing waste liquid specifically comprises the following steps: s1, selecting and weighing raw materials: firstly, weighing ASP dry powder with required weight by a weighing device, transferring the weighed ASP dry powder into a storage tank for storage, and preparing for use, wherein S2, dissolving raw materials: pouring the ASP dry powder weighed in the step S1 into a dissolving bottle, then adding water with a corresponding volume into the dissolving bottle, stirring for 30-40min by a stirring rod to completely dissolve the ASP dry powder, and S3, desalting the solution: transferring the solution dissolved in the step S2 to a desalting apparatus, filtering the dissolved macromolecular salt through an RO reverse osmosis desalination membrane by using mechanical pressure, and allowing the filtered solution to flow into a collection tank for concentrated collection treatment, thereby completing desalting treatment, S4, chromatography and purification treatment of the solution: transferring the desalted solution obtained in the step S3 to a purification device, performing chromatography on the solution by using DEAE ion exchange chromatography, then purifying and collecting BBI protein and KTI protein, S5, concentrating and mixing with an ultrafiltration membrane: adjusting the pH of the solution purified in the step S4 to 5-6, concentrating by using an ultrafiltration membrane, adding the macromolecular substances subjected to the ultrafiltration membrane and the BBI protein and the KTI protein collected after purification in the step S4 into a mixing device for mixing treatment, and S6, preparing an SBTI finished product: the mixed liquid is transferred into a filtering device after being mixed in the step S5, the mixed liquid is filtered by a filter membrane, the filtered substance is dried at the temperature of 45-55 ℃, an SBTI finished product is obtained after drying, the ASP dry powder extracted from soybean clear wastewater can be used for directly carrying out SBTI refining treatment, the aim of rapidly extracting SBTI is well fulfilled by recycling soybean deep processing waste liquid, the SBTI refining cost is saved, the SBTI production efficiency is improved, the SBTI refining purity is improved by directly refining the ASP dry powder, the SBTI refining efficiency is greatly improved, the production cost is saved, and the SBTI refining is very beneficial to the SBTI refining.

Drawings

FIG. 1 is a flow chart of the present invention.

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, the embodiment of the present invention provides four technical solutions: a process for refining SBTI by ASP dry powder based on soybean deep processing waste liquid specifically comprises the following embodiments: