A bacterial expression system revisited for the recombinant production of cystine-rich plant lipid transfer proteins
A bacterial expression system revisited for the recombinant production of cystine-rich plant lipid transfer proteins
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DOI：
10.1016/j.bbrc.2004.02.173
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发表时间：
2004-04-16
影响因子：
3.1
通讯作者：
Marion, D
中科院分区：
文献类型：
--
作者：
Elmorjani, K;Lurquin, V;Marion, D
非特异性脂质转移蛋白（nsLTPs）是一种丰富且普遍存在的富含半胱氨酸的蛋白质，在体外能够结合脂质和疏水分子。为了探究小麦nsLTP1的脂质结合特性，突变变体可能是一个强有力的工具。为此，设计了一个编码成熟小麦nsLTP1多肽的合成基因，以确保在大肠杆菌中高水平表达。优化了细菌表达宿主菌株、翻译融合策略以及方便的裂解和纯化程序，在标准发酵条件下生产了大量（最终产量为15 mg/L）可溶且正确折叠的重组nsLTP1。该表达系统具有高度的亲和性，有助于研究植物nsLTP的构效关系。(C) 2004爱思唯尔公司版权所有。
Non-specific lipid transfer proteins (nsLTPs) are abundant and ubiquitous cystine-rich proteins that are capable, in vitro, of binding lipids and hydrophobic molecules. In view to probe the lipid binding properties of the wheat nsLTP1, mutant variants may represent a powerful tool. To this end, a synthetic gene, encoding a mature wheat nsLTP1 polypeptide, was designed to ensure high level expression in Escherichia coli. The bacterial expression host strain, a translational fusion strategy, and convenient cleavage and purification procedures were optimized to produce in standard fermentation conditions, a significant amount (15 mg/L final yield), of a soluble and correctly folded recombinant nsLTP1. This highly amenable expression system is helpful in order to investigate structure-activity relationships of plant nsLTP. (C) 2004 Elsevier Inc. All rights reserved.