کد مقاله کد نشریه سال انتشار مقاله انگلیسی ترجمه فارسی نسخه تمام متن
6452317 1417010 2016 8 صفحه PDF سفارش دهید دانلود رایگان
عنوان انگلیسی مقاله ISI
Pichia pastoris mutants as host strains for efficient secretion of recombinant branched chain aminotransferase (BCAT)
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موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
Pichia pastoris mutants as host strains for efficient secretion of recombinant branched chain aminotransferase (BCAT)
چکیده انگلیسی


- Escherichia coli BCAT has been expressed as a secreted protein in the supernatant of Pichia pastoris for the first time.
- Novel Pichia pastoris strains (Δcwp1, Δscw10) have been generated as a strategy to improve protein production.
- Improved BCAT secretion by OCH1 knock-out in Pichia pastoris, providing a promising system for efficient chiral amine production.

Branched chain aminotransferase (BCAT) is one of the enzymatic tools of choice for the production of chiral amines or amino acids; especially, non-natural amino acids are of interest as building blocks for the pharmaceutical industry. The expression and subsequent secretion of BCAT counteracts limited cell permeability of target substrates and facilitates downstream processing. Since Pichia pastoris secretes a negligible amount of native proteins and was previously shown to efficiently secrete recombinant proteins, it was chosen as the expression host. We examined different promoters and glycosylation states and also engineered the host strain by disrupting genes encoding proteins related to cell wall assembly (Scw10, Cwp1) and glycosylation (Och1). Finally, we were able not only to increase the extracellular BCAT production, but also to achieve a more homogenous product in terms of glycosylation and identified a deletion strain, which counteracts typical cell clustering in the Δoch1 strain.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 235, 10 October 2016, Pages 84-91
نویسندگان
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