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Book synopsis

This groundbreaking work unveils a paradigm-shifting platform for in situ protein assembly and precision purification, merging split intein engineering with microbial consortia design. Pioneering a ""one-pot"" biosynthesis strategy, the book details how engineered Escherichia coli equipped with the ePop autolysis system synchronously releases intein-tagged protein fragments, while Pichia pastoris secretes complementary domains via optimized eukaryotic secretion pathways. By integrating quorum-sensing controls and split intein-mediated trans-splicing, this system achieves extracellular protein reconstitution directly in culture, bypassing costly in vitro purification steps . The platform's modularity enables logic-gated synthesis of multi-domain proteins, exemplified by its applications in antibiotic resistance engineering and context-responsive biomaterials.
Central to this innovation is the rational engineering of the Mycobacterium xenopi GyrA intein. Through targeted mutagenesis (Cys1Ala, Ser179Gly, His197Gln), we developed a pH/calcium-responsive self-cleaving tag that releases bioactive human basic fibroblast growth factor (bFGF) with >85% efficiency. The Ser179Gly mutant exhibits rapid C-terminal cleavage under alkaline conditions (50 mM Tris-HCl, pH 8.5), while the Cys1Ala/His197Gln double mutant boosts precursor yield 3.2-fold-a breakthrough for high-titer production. A novel 3-step purification cascade (cation exchange → heparin affinity → gel filtration) recovers 85% of bioactive bFGF from lysates, validated by LC-MS/MS and MTT assays. Parallel protocols for human epidermal growth factor (hEGF) and growth hormone (hGH) demonstrate platform versatility.

Book details

Book category Books in German Naturwissenschaften, Medizin, Informatik, Technik Technik Maschinenbau, Fertigungstechnik

• Full title: Intein
• Author: Hao Wang
• Language: English
• Binding: Paperback
• Number of pages: 174
• EAN: 9789999327091
• ISBN: 9999327095
• ID: 50101877
• Publisher: Eliva Press
• Weight: 262 g
• Dimensions: 229 × 152 × 10 mm
• Date of publishing: 07. November 2025

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