Polyhydroyxalkanoate Synthase Fusions as a Strategy for Oriented Enzyme Immobilisation

Molecules

David O. Hooks¹  Mark Venning-Slater¹  Jinping Du¹ 

[1] Institute of Fundamental Sciences, Massey University, Private Bag 11222, Palmerston North 4442, New Zealand; E-Mails:

关键词: Ralstonia eutropha;    polyhydroxyalkanoate synthase;    enzyme immobilisation;   

DOI  :  10.3390/molecules19068629

来源: mdpi

【 摘 要 】

Polyhydroxyalkanoate (PHA) is a carbon storage polymer produced by certain bacteria in unbalanced nutrient conditions. The PHA forms spherical inclusions surrounded by granule associate proteins including the PHA synthase (PhaC). Recently, the intracellular formation of PHA granules with covalently attached synthase from Ralstonia eutropha has been exploited as a novel strategy for oriented enzyme immobilisation. Fusing the enzyme of interest to PHA synthase results in a bifunctional protein able to produce PHA granules and immobilise the active enzyme of choice to the granule surface. Functionalised PHA granules can be isolated from the bacterial hosts, such as Escherichia coli, and maintain enzymatic activity in a wide variety of assay conditions. This approach to oriented enzyme immobilisation has produced higher enzyme activities and product levels than non-oriented immobilisation techniques such as protein inclusion based particles. Here, enzyme immobilisation via PHA synthase fusion is reviewed in terms of the genetic designs, the choices of enzymes, the control of enzyme orientations, as well as their current and potential applications.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.