Construction of a chassis for hydrogen production: physiological and molecular characterization of a Synechocystis sp PCC 6803 mutant lacking a functional bidirectional hydrogenase

• Type: Article in International Scientific Journal
• Date: 2012

Authors

Filipe Pinto (Author) Other; Karin A van Elburg (Author) Other; Catarina C Pacheco (Author) Other; Miguel Lopo (Author) Other; Josselin Noirel (Author) Other; Arnau Montagud (Author) Other; Javier F Urchueguia (Author) Other; Phillip C Wright (Author) Other; Paula Tamagnini (Author) FEUP

Journal

Title: Microbiology

Vol. 158

Pages: 448-464

ISSN: 1350-0872

Publisher: Microbiological Society

Indexing

ISI Web of Knowledge - 21 Citations

Scopus - 24 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Other information

Authenticus ID: P-002-DCM

DOI: 10.1099/mic.0.052282-0

Abstract (EN): Cyanobacteria are photosynthetic prokaryotes that are promising 'low-cost' microbial cell factories due to their simple nutritional requirements and metabolic plasticity, and the availability of tools for their genetic manipulation. The unicellular non-nitrogen-fixing Synechocystis sp. PCC 6803 is the best studied cyanobacterial strain and its genome was the first to be sequenced. The vast amount of physiological and molecular data available, together with a relatively small genome, makes Synechocystis suitable for computational metabolic modelling and to be used as a photoautotrophic chassis in synthetic biology applications. To prepare it for the introduction of a synthetic hydrogen producing device, a Synechocystis sp. PCC 6803 deletion mutant lacking an active bidirectional hydrogenase (Delta hoxYH) was produced and characterized at different levels: physiological, proteomic and transcriptional. The results showed that, under conditions favouring hydrogenase activity, 17 of the 210 identified proteins had significant differential fold changes in comparisons of the mutant with the wild-type. Most of these proteins are related to the redox and energy state of the cell. Transcriptional studies revealed that only six genes encoding those proteins exhibited significant differences in transcript levels. Moreover, the mutant exhibits similar growth behaviour compared with the wild-type, reflecting Synechocystis plasticity and metabolic adaptability. Overall, this study reveals that the Synechocystis Delta hoxYH mutant is robust and can be used as a photoautotrophic chassis for the integration of synthetic constructs, i.e. molecular constructs assembled from well characterized biological and/or synthetic parts (e.g. promoters, regulators, coding regions, terminators) designed for a specific purpose.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 17