Multi-Enzyme Assembly on T4 Phage Scaffold

Frontiers in Bioengineering and Biotechnology
Jinny L LiuEllen R Goldman

Abstract

Over the past two decades, various scaffolds have been designed and synthesized to organize enzyme cascades spatially for enhanced enzyme activity based on the concepts of substrate channeling and enhanced stability. The most bio-compatible synthetic scaffolds known for enzyme immobilization are protein and DNA nanostructures. Herein, we examined the utility of the T4 phage capsid to serve as a naturally occurring protein scaffold for the immobilization of a three-enzyme cascade: Amylase, Maltase, and Glucokinase. Covalent constructs between each of the enzymes and the outer capsid protein Hoc were prepared through SpyTag-SpyCatcher pairing and assembled onto phage capsids in vitro with an estimated average of 90 copies per capsid. The capsid-immobilized Maltase has a fourfold higher initial rate relative to Maltase free in solution. Kinetic analysis also revealed that the immobilized three-enzyme cascade has an 18-fold higher converted number of NAD+ to NADH relative to the mixtures in solution. Our results demonstrate that the T4 phage capsid can act as a naturally occurring scaffold with substantial potential to enhance enzyme activity by spatially organizing enzymes on the capsid Hoc.

References

Apr 23, 1999·The Journal of Biological Chemistry·E W MilesD R Davies
Apr 9, 2004·Proceedings of the National Academy of Sciences of the United States of America·Andrei FokineMichael G Rossmann
Aug 30, 2008·Current Opinion in Biotechnology·Robert J ConradoMatthew P DeLisa
Aug 4, 2009·Nature Biotechnology·John E DueberJay D Keasling
Mar 9, 2010·Journal of Biological Engineering·J Christopher AndersonJay D Keasling
Dec 7, 2010·Virology Journal·Venigalla B Rao, Lindsay W Black
Jun 16, 2011·Biotechnology Advances·Y-H Percival Zhang
Jan 31, 2012·Biochemical and Biophysical Research Communications·Kelly RobertsonJinny L Liu
Mar 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Bijan ZakeriMark Howarth
May 9, 2012·Trends in Biotechnology·Daniela CardinaleThierry Michon
Jun 16, 2012·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Kelly L Robertson, Jinny L Liu
Aug 28, 2014·Proceedings of the National Academy of Sciences of the United States of America·Jinny L LiuLindsay W Black
Sep 1, 2015·Current Opinion in Biotechnology·Ka-Hei SiuWilfred Chen
Oct 31, 2015·Current Opinion in Chemical Biology·Samuel C Reddington, Mark Howarth
May 6, 2016·Nature Nanotechnology·Andreas KüchlerPeter Walde
Jan 13, 2018·Heliyon·Ellen R GoldmanJinny L Liu
Dec 18, 2018·Frontiers in Bioengineering and Biotechnology·Mathilde KochOlivier Borkowski

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Methods Mentioned

BETA
atomic force microscopy
size exclusion chromatography

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