کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10843784 | 1069293 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Cloning and expression of earthworm fibrinolytic enzyme PM246 in Pichia pastoris
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
We have cloned, expressed, and purified a novel earthworm fibrinolytic enzyme (EFE) of Lumbricus rubellus in Pichia pastoris. Its cDNA sequence (GenBank Accession No. AY178854) revealed a 747Â bp region containing an intact ORF that encodes a protein of 246 amino acid residues, designated as EFE PM246. While EFE PM246 is distinct, its cDNA shows a high degree of sequence homologies with four other EFE cDNAs registered in GenBank. The recombinant EFE PM246 was active, showing a fibrinolytic activity of 7.5Â ÃÂ 106Â U/L in basal salts medium, a higher fibrinolytic activity than those produced in other expression systems. The recombinant EFE PM246 expressed in basal salts medium was purified by a three-step purification procedure with a recovery rate of about 20%. This is the first report detailing the successful purification of a genetically engineered earthworm fibrinolytic enzyme. The main physiochemical features of the EFE PM246, including temperature stability, pH resistance, and sensitivity to some protein inhibitors, were also characterized.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 43, Issue 1, September 2005, Pages 18-25
Journal: Protein Expression and Purification - Volume 43, Issue 1, September 2005, Pages 18-25
نویسندگان
Yan Hu, Xiao-Lin Meng, Jin-Ping Xu, Wei Lu, Jian Wang,