کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
6452773 1418339 2017 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Original Research ArticleOvercoming the membrane barrier: Recruitment of γ-glutamyl transferase for intracellular release of metabolic cargo from peptide vectors
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
Original Research ArticleOvercoming the membrane barrier: Recruitment of γ-glutamyl transferase for intracellular release of metabolic cargo from peptide vectors
چکیده انگلیسی


- A synthetic transport system to overcome the membrane barrier of E. coli.
- Intracellular release of cargo molecules using γ-glutamyl transferase (GGT).
- Expression of GGT in the cytoplasm of E. coli leads to toxic effects.
- Toxic effects can be mitigated through ribosome binding site engineering.

Semipermeable membranes of cells frequently pose an obstacle in metabolic engineering by limiting uptake of substrates, intermediates, or xenobiotics. Previous attempts to overcome this barrier relied on the promiscuous nature of peptide transport systems, but often suffered from low versatility or chemical instability. Here, we present an alternative strategy to transport cargo molecules across the inner membrane of Escherichia coli based on chemical synthesis of a stable cargo-peptide vector construct, transport through the peptide import system, and efficient intracellular release of the cargo by the promiscuous enzyme γ-glutamyl transferase (GGT). Retaining the otherwise periplasmic GGT in the cytoplasm was critical for the functionality of the system, as was fine-tuning its expression in order to minimize toxic effects associated to cytoplasmic GGT expression. Given the established protocols of peptide synthesis and the flexibility of peptide transport and GGT, the system is expected to be suitable for a broad range of cargoes.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Metabolic Engineering - Volume 39, January 2017, Pages 60-70
نویسندگان
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