Cell-free freeze-drying preparation for in-vitro protein synthesis, and preparation method and application of cell-free freeze-drying preparation

Abstract

The invention provides a cell-free freeze-drying preparation for in-vitro protein synthesis, and a preparation method and application of the cell-free freeze-drying preparation. Specifically, throughthe cell-free freeze-drying preparation provided by the invention, the synthetic activity of exogenous protein can be significantly improved under the situation that a protective agent does not need to be added.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a cell-free freeze-dried preparation for protein synthesis in vitro, its preparation method and application. Background technique [0002] Vacuum freeze-drying technology is to freeze the wet material or solution into a solid state at a lower temperature (-10 ° C ~ -50 ° C), and then sublimate the water in it directly into a gaseous state without going through a liquid state under vacuum (1.3 ~ 13 Pa). , the drying technology that finally dehydrates the material. The whole process of vacuum freeze-drying is divided into pre-freezing, sublimation drying in the first stage and re-drying in the second stage. [0003] At present, the freeze-drying method is an effective and commonly used method to maintain protein activity, which is beneficial to storage and transportation, but there are some potential damages in the freeze-drying process[1]. During the process of pre-cooling, primary ...

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Problems solved by technology

[0005] One disadvantage of the cell-free expression system is the preservation of cell extracts. The liquid must be stored at -80°C and cannot be frozen and thawed repeatedly, otherwise it will seriously affect the ability of cell extracts to synthesize proteins. Keep the liquid cell extract solution at room temperature. Its protein synthesis capacity will decrease over time until it becomes inactive[4]

[0006] However, there are still no successful cases of freeze-drying other eukaryotic cells, such as yeast, cell-free expression systems

[0007] At present, the freeze-drying process of the cell extract of the yeast in vitro protein synthesis system has not been reported. In addition to the cell extract, the in vitro protein synthesis system also contains amino acids, NTP, buffer, RNA polymerase and other necessary components for protein synthesis. Some of the components need to be stored at low or extremely low temperatures, which greatly increases the difficulty and cost of storage and transportation, and requires repeated freezing and thawing during use, which affects the activity of protein synthesis. It is inconvenient to operate and not suitable for large-scale Industrial production and use

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