Liquid Marbles as an Easy‐to‐Handle Compartment for Cell‐Free Synthesis and In Situ Immobilization of Recombinant Proteins. Issue 12 (10th September 2018)

• Record Type: Journal Article
• Authors: Kamiya, Noriho; Ohama, Yuki; Minamihata, Kosuke; Wakabayashi, Rie; Goto, Masahiro
• Is Part Of: Biotechnology journal. Volume 13:Issue 12(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-09-10
• Subjects: cell‐free protein synthesis -- horseradish peroxidase -- hydrogels -- liquid marble -- protein immobilization; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201800085
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract : Liquid marble (LM), a self‐standing micro‐scale aqueous droplet, emerges as a micro‐bioreactor in biological applications. Herein, the potential of LM as media for cell‐free synthesis and simultaneous immobilization of recombinant proteins is explored. Initially, formation of hydrogel marble (HM) by using an enzymatic disulfide‐based hydrogelation technique is confirmed by incorporating three components, horseradish peroxidase (HRP), a tetra‐thiolated poly(ethylene glycol) derivative, and glycyl‐L‐tyrosine, in LM. The compatibility of the enzymatic hydrogelation with cell‐free protein synthesis in LM is then validated. Although the hydrogelation reduces the level of protein synthesis in LM when compared with that in a test tube, the biosynthesis of enhanced green fluorescent protein (EGFP) is achieved. Interestingly, EGFP synthesized in LM is entrapped in the HM, and the introduction of a cysteine residue to EGFP by genetic engineering further increases the amount of protein immobilization in the hydrogel matrices. These results suggest that the cell‐free synthesis and HRP‐catalyzed hydrogelation can be conducted in parallel in LM, and the eventual entrapment of the key components in HM is possible. Facile recovery of macromolecular products immobilized in HM by degrading the hydrogel network under reducing conditions should lead to the design of an easy‐to‐handle system to screen protein functions. Abstract : The potential of liquid marble (LM) as a micro‐sized self‐standing compartment for cell‐free protein synthesis (CFPS) and in situ immobilization of biosynthesized proteins in hydrogel marble (HM) are validated. Ease of recovery of macromolecular products entrapped in HM by degrading the hydrogel network under reducing conditions leads to an easy‐to‐handle screening system of protein functions. This article is part of an AFOB (Asian Federation of Biotechnology) Special issue. To learn more about the AFOB visitwww.afob.org .