Intein: The Molecular Revolution of Protein Splicing: From Mechanistic Insights to Biotechnological Applications...

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Hao Wang

Intein

The Molecular Revolution of Protein Splicing: From Mechanistic Insights to Biotechnological...

Hao Wang

$137.95 $110.63

Paperback

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English

Eliva Press
07 November 2025

Technology: general issues

Summary
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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.

By: Hao Wang
Imprint: Eliva Press
Dimensions: Height: 229mm, Width: 152mm, Spine: 9mm
Weight: 242g
ISBN: 9789999327091
ISBN 10: 9999327095
Pages: 174
Publication Date: 07 November 2025
Audience: General/trade , ELT Advanced
Format: Paperback
Publisher's Status: Active

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Intein

The Molecular Revolution of Protein Splicing: From Mechanistic Insights to Biotechnological...

Hao Wang

$137.95 $110.63

Paperback

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