Complex waste stream valorisation through combined enzymatic hydrolysis and catabolic assimilation byPseudomonas putida

Abstract

Linear consumption models in combination with the poor waste management results in negative impact upon the climate and environment. Here we show an advanced biorefinery concept to produce biopolymers and recombinant therapeutic proteins from municipal solid waste. Careful selection and metabolic engineering of Pseudomonas putida was employed to permit efficient co-utilisation of complex, heterogenous substrate compositions derived from the most abundant municipal solid waste fractions. Enzymatic pre-treatment of the feedstock was performed, which afforded sufficient carbon for growth and bio-production of value-added products. Unlike current waste processing methods and supply of feedstocks for bio-based manufacturing, this biorefinery concept re-directs the carbon away from direct emissions and creates a land-use negative feedstock ideal for advanced biomanufacturing. We envisage this demonstration will lead to the creation of circular consumption solutions for waste and bio-production of a range of chemicals and materials in future resource finite scenarios.

Tuning the performance of a TphR-based terephthalate biosensor with a design of experiments approach

“…Esterases from Clostridium thermocellum and Cellvibrio japonicus were purchased from Prozomix Limited, lipases from Pseudomonas fluorescens , Aspergillus niger , Candida rigosa , Mucor miehei , Burkholderia cepacia , Rhizopus niveus , Candida antarctica (CALB), and porcine liver were purchased from Merck. The LCC-ICCG cutinase was produced and purified as previously described 40 .…”

“…TPA is an aroma;c building block of polyethylene terephthalate (PET) plas;c, and is released aYer enzyma;c or chemical deconstruc;on of PET [32][33][34] . Therefore, op;mal TPA detec;on methods find immediate applica;on in the discovery and engineering of novel PET hydrolases, or the dynamic regula;on of biosynthe;c pathways towards PET valorisa;on [35][36][37][38][39][40] . Here, we iden;fied and characterized novel regulatory components to build a fully modularized TPA biosensor design, which we then used to build an efficient TPA-responsive promoter library that covers a wide range of sensing strengths, dynamic ranges, and sensi;vi;es.…”