期刊
BIOPOLYMERS
卷 106, 期 6, 页码 818-824出版社
WILEY
DOI: 10.1002/bip.22875
关键词
sunflower trypsin inhibitor; Bowman-Birk inhibitor; backbone-cyclized peptides; protein splicing; split-intein
资金
- National Institutes of Health Research Grants [R01-GM090323, R01-GM113363, R01-GM085006]
We report for the first time the recombinant expression of bioactive wild-type sunflower trypsin inhibitor 1 (SFTI-1) inside E. coli cells by making use of intracellular protein trans-splicing in combination with a high efficient split-intein. SFTI-1 is a small backbone-cyclized polypeptide with a single disulfide bridge and potent trypsin inhibitory activity. Recombinantly produced SFTI-1 was fully characterized by NMR and was observed to actively inhibit trypsin. The in-cell expression of SFTI-1 was very efficient reaching intracellular concentration approximate to 40 mM. This study clearly demonstrates the possibility of generating genetically encoded SFTI-based peptide libraries in live E. coli cells, and is a critical first step for developing in-cell screening and directed evolution technologies using the cyclic peptide SFTI-1 as a molecular scaffold. (C) 2016 Wiley Periodicals, Inc.
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