Barnacle glue protein dialysis process and method for acquiring two different types of proteins

A gelatin and barnacle technology, which is applied to the dialysis process of biological materials and the field of obtaining protein, can solve the problems of low colloidal protein yield and long dialysis time, and achieves the advantages of shortening dialysis time, increasing application scope and high gelation rate. Effect

Active Publication Date: 2019-04-16
NAT UNIV OF DEFENSE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the output of colloidal protein in the above-mentioned patent is less, and the dialysis time is longer

Method used

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  • Barnacle glue protein dialysis process and method for acquiring two different types of proteins
  • Barnacle glue protein dialysis process and method for acquiring two different types of proteins
  • Barnacle glue protein dialysis process and method for acquiring two different types of proteins

Examples

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Effect test

Embodiment 1

[0034] A Trx-Balcp19k protein dialysis process, comprising the following steps: putting the Trx-Balcp19k protein solution with a concentration of 2.11 mg / mL and a volume of 14 mL obtained by affinity chromatography (see patent CN105031717A) into a dialysis bag , under magnetic stirring, placed in an environment of 4°C for dialysis with deionized water as the dialysate, during the dialysis process, the dialysate was replaced every 2 hours, and the dialysate was continued for 8 hours (the ionic strength in the dialysate at this time was 173 μs / cm ). Such as figure 1As shown, as the dialysis time prolongs, viscous jelly will appear on the inner wall of the dialysis bag, and it is mainly concentrated in the middle and lower part of the dialysis bag. It should be noted that dialysis itself will lose a part of protein, and according to past experience, the loss rate is about 30%.

[0035] In this embodiment, after the dialysate was replaced for the first time, the protein solution...

Embodiment 2

[0043] A Trx-Balcp19k protein dialysis process, comprising the following steps: putting the Trx-Balcp19k protein solution with a concentration of 2.11 mg / mL and a volume of 14 mL obtained by affinity chromatography (see patent CN105031717A) into a dialysis bag , under magnetic stirring, place in an environment of 4°C and use deionized water as the dialysate for dialysis, and change the dialysate every 2 hours during the dialysis process (the time for changing the dialysate needs to meet the requirements of the gelation rate at the same time, the same below) , and after 4 hours of dialysis, the viscous jelly was collected from the inner wall of the dialysis bag with the tip of a pipette gun, and the dialysis was continued for 8 hours (at this time, the ionic strength in the dialysate was 149.3 μs / cm). Such as image 3 As shown, as the dialysis time prolongs, viscous jelly will appear on the inner wall of the dialysis bag, and it is mainly concentrated in the middle and lower pa...

Embodiment 3

[0050] A Trx-Balcp19k protein dialysis process, comprising the following steps: putting the Trx-Balcp19k protein solution with a concentration of 2.37 mg / mL and a volume of 15 mL obtained by affinity chromatography (see patent CN105031717A) into a dialysis bag , under magnetic stirring, placed in an environment of 4°C for dialysis with deionized water as the dialysate, during the dialysis process the dialysate was replaced every 2 hours, and after 4 hours of dialysis, use the tip of a pipette to collect the viscous glue from the inner wall of the dialysis bag Shaped matter, continuous dialysis for 8h.

[0051] In this embodiment, after the dialysate was replaced for the first time, the protein solution concentration was about 1.73mg / mL, the volume was 16.3mL, and the gelation rate was about 7.8%; after the dialysate was replaced for the second time, the protein solution concentration was about 1.32 mg / mL, the volume is 17.1mL, the gelation rate is about 19.5%; after the third ...

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Abstract

The invention discloses a barnacle glue protein dialysis process. The barnacle glue protein dialysis process comprises the following steps that a barnacle glue protein solution is added in a dialysisbag and dialysed with deionized water as a dialysate; during dialysis, the dialyste is magnetically stirred in the whole process; the dialysate is replaced multiple times during dialysis; during dialysis, colloidal proteins on the inner wall of the dialysis bag are continually collected until dialysis is completed. The invention further correspondingly provides a method for simultaneously collecting two differently types of proteins from a protein solution containing Trx-Balcp19k. According to the barnacle glue protein dialysis process, the replacement frequency and occasion of the dialysate are controlled, the colloidal proteins on the inner wall of the dialysis bag are collected in time so that a larger protein adsorption interface can be provided and more viscous colloidal proteins canbe obtained, and thus the gelatinize rate is higher. In addition, by the adoption of the barnacle glue protein dialysis process, the dialysis time is obviously shortened and can be shortened from original 12 h to 8-10 h under the condition of the same gelatinize rate.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and in particular relates to a dialysis process of biological materials and a method for obtaining protein. Background technique [0002] Strong fouling biological barnacles in the ocean adhere and solidify on the surface of underwater objects by secreting proteinaceous biological glue (barnacle glue), achieving permanent attachment and fouling. Barnacle glue is a multi-protein complex, which relies on the interaction of multiple proteins to exert a strong adhesion ability in complex water environments. This adhesion model provides new inspiration for human beings to study underwater strong adhesives, and has important application value in the fields of new antifouling technology, underwater repair and biomedicine. [0003] In patent CN105031717A, the recombinant barnacle glue protein cp19k obtained by Escherichia coli heterologous expression can obtain a kind of jelly with good viscosity thr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/34
CPCC07K14/43509
Inventor 胡碧茹曾玲景承宇叶宗煌刘志明吴文健解江冰
Owner NAT UNIV OF DEFENSE TECH
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