在黑曲霉中表达并分泌的鸡蛋清溶菌酶能被正确加工和折叠。

Hen egg white lysozyme expressed in, and secreted from, Aspergillus niger is correctly processed and folded.

作者信息

Archer D B, Jeenes D J, MacKenzie D A, Brightwell G, Lambert N, Lowe G, Radford S E, Dobson C M

机构信息

Agricultural and Food Research Council Institute of Food Research, Norwich Laboratory, U.K.

出版信息

Biotechnology (N Y). 1990 Aug;8(8):741-5. doi: 10.1038/nbt0890-741.

DOI:10.1038/nbt0890-741

PMID:1366900

Abstract

We transformed Aspergillus niger with the full length cDNA gene encoding hen egg-white lysozyme (HEWL) and its secretion signal sequence. Lysozyme levels up to 12 mg/l were secreted when expression was controlled by the A. awamori glucoamylase (GAM) promoter and 1 mg/l when controlled by the A. nidulans glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter. N-terminal sequence analysis of the recombinant protein indicated that the signal peptide was correctly processed by the A. niger secretory apparatus. The specific catalytic activity of the recombinant protein was identical to that of authentic hen lysozyme. The recombinant HEWL was examined by 2D 1H-NMR spectroscopy and shown to have a spectrum identical to that of authentic HEWL indicating that the protein was correctly folded.

摘要

我们用编码鸡蛋清溶菌酶（HEWL）及其分泌信号序列的全长cDNA基因转化了黑曲霉。当表达由泡盛曲霉糖化酶（GAM）启动子控制时，溶菌酶分泌水平高达12 mg/l；当由构巢曲霉甘油醛-3-磷酸脱氢酶（GPD）启动子控制时，分泌水平为1 mg/l。重组蛋白的N端序列分析表明信号肽被黑曲霉分泌装置正确加工。重组蛋白的比催化活性与天然鸡蛋清溶菌酶相同。通过二维1H-NMR光谱对重组HEWL进行检测，结果显示其光谱与天然HEWL的光谱相同，表明该蛋白折叠正确。