eprintid: 24304
rev_number: 12
eprint_status: archive
userid: 1589
dir: disk0/00/02/43/04
datestamp: 2018-04-25 08:08:05
lastmod: 2024-03-11 04:03:09
status_changed: 2018-04-25 08:08:05
type: article
metadata_visibility: show
creators_name: Du, Wei
creators_name: Jongbloets, Joeri A.
creators_name: van Boxtel, Coco
creators_name: Hernández, Hugo Pineda
creators_name: Lips, David
creators_name: Oliver, Brett G.
creators_name: Hellingwerf, Klaas J.
creators_name: Branco dos Santos, Filipe
title: Alignment of microbial fitness with engineered product formation: obligatory coupling between acetate production and photoautotrophic growth
subjects: ddc-570
divisions: i-716000
abstract: Background: Microbial bioengineering has the potential to become a key contributor to the future development of human society by providing sustainable, novel, and cost-effective production pipelines. However, the sustained productivity of genetically engineered strains is often a challenge, as spontaneous non-producing mutants tend to grow faster and take over the population. Novel strategies to prevent this issue of strain instability are urgently needed.

Results: In this study, we propose a novel strategy applicable to all microbial production systems for which a genome-scale metabolic model is available that aligns the production of native metabolites to the formation of biomass. Based on well-established constraint-based analysis techniques such as OptKnock and FVA, we developed an in silico pipeline—FRUITS—that specifically ‘Finds Reactions Usable in Tapping Side-products’. It analyses a metabolic network to identify compounds produced in anabolism that are suitable to be coupled to growth by deletion of their re-utilization pathway(s), and computes their respective biomass and product formation rates. When applied to Synechocystis sp. PCC6803, a model cyanobacterium explored for sustainable bioproduction, a total of nine target metabolites were identified. We tested our approach for one of these compounds, acetate, which is used in a wide range of industrial applications. The model-guided engineered strain shows an obligatory coupling between acetate production and photoautotrophic growth as predicted. Furthermore, the stability of acetate productivity in this strain was confirmed by performing prolonged turbidostat cultivations.

Conclusions: This work demonstrates a novel approach to stabilize the production of target compounds in cyanobacteria that culminated in the first report of a photoautotrophic growth-coupled cell factory. The method developed is generic and can easily be extended to any other modeled microbial production system.
date: 2018
publisher: BioMed Central
id_scheme: DOI
ppn_swb: 165526267X
own_urn: urn:nbn:de:bsz:16-heidok-243043
language: eng
bibsort: DUWEIALIGNMENTO2018
full_text_status: public
publication: Biotechnology for biofuels
volume: 11
number: 38
place_of_pub: London
pagerange: 1-13
issn: 1754-6834
citation:   Du, Wei ; Jongbloets, Joeri A. ; van Boxtel, Coco ; Hernández, Hugo Pineda ; Lips, David ; Oliver, Brett G. ; Hellingwerf, Klaas J. ; Branco dos Santos, Filipe  (2018) Alignment of microbial fitness with engineered product formation: obligatory coupling between acetate production and photoautotrophic growth.  Biotechnology for biofuels, 11 (38).  pp. 1-13.  ISSN 1754-6834     
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/24304/1/13068_2018_Article_1037.pdf