Molecular cloning, nucleotide sequence and expression of the gene encoding prepro‐polygalacturonasell of Aspergillus niger

FEBS Letters

Kester, Harry C.M.¹  Visser, Jaap¹  Bussink, Hendrik J.D.¹ 

[1] Department of Genetics, Agricultural University, Dreyenlaan 2,6703 HA Wageningen, The Netherlands

关键词: Prepro-peptide processing;    Polygalacturonase;    Transformation;    Industrial enzyme;    Secretion;    Aspergillus niger;    PG;    polygalacturonase;    pgaII;    the PGII gene;    bp;    base pair;    DABITC;    dimethylaminoazobenzene isothiocyanate;   

DOI  :  10.1016/0014-5793(90)81066-W

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

PolygalacturonaseII of Aspergillus niger was fragmented using CNBr and the NH₂-terminal fragment and another fragment were partially sequenced. The polygalacturonaseII (pgaII) gene was then isolated by using an oligonucleotide mixture based on the internal amino acid sequence as a probe. The nucleotide sequence of the pga II structural gene was determined. It was found that polygalacturonaseII is synthesized as a precursor having an NH₂-terminal prepro-sequence of 27 amino acids. The cloned gene was used to construct polygalacturonaseII over-producing A. niger strains. PolygalacturonaseII was isolated from one such strain and was determined to be correctly processed and to be fully active.

【 授权许可】

Unknown