一种无需裂解细胞的蛋白提取方法

By constructing the fusion enzyme CPP-T4L, protein extraction without cell lysis was achieved, solving the problems of cumbersome operation and high cost in traditional methods, and improving production efficiency and economic benefits.

CN112143748BActive Publication Date: 2026-07-17SHAANXI UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SHAANXI UNIV OF SCI & TECH
Filing Date
2020-10-13
Publication Date
2026-07-17

AI Technical Summary

Technical Problem

Traditional protein extraction methods require a cell lysis step, which is cumbersome and requires sophisticated equipment, resulting in low production efficiency and increased costs.

Method used

By constructing the fusion enzyme CPP-T4L, its membrane-penetrating and cell wall-degrading properties are utilized to induce the expression of the target protein after expression, enabling protein extraction without cell lysis and simplifying the operation process.

Benefits of technology

It eliminates the cell recovery and disruption steps, significantly reducing production costs, improving production efficiency, simplifying the operation process, and achieving high protein extraction efficiency.

✦ Generated by Eureka AI based on patent content.

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Abstract

本发明提供一种无需裂解细胞的蛋白提取方法，包括：将穿膜肽CPP基因与细菌裂解酶T4L基因通过柔性连接肽基因联合，形成融合酶CPP‑T4L基因，并连接到融合酶表达载体中，再将融合酶表达载体转入表达菌株中，得到重组宿主菌株；将目标蛋白表达基因克隆到目标蛋白表达载体上，得到目的蛋白表达载体，再将其转化至重组宿主菌株中，构建重组表达菌株；在重组表达菌株的表达体系中，先诱导目标蛋白的表达；目标蛋白表达完成后，添加IPTG启动融合酶的表达，释放目标蛋白；过滤除去细胞残渣后，收集细胞裂解上清液，回收目标蛋白。本发明省去了传统表达方法中细胞回收及破碎的步骤，极大简化了操作流程。
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