Growth-dependent recombinant product formation kinetics can be reproduced through engineering of glucose transport and is prone to phenotypic heterogeneity

Autor: Frank Delvigne, Alfredo Martinez, Thai Minh Nguyen, Wenzheng Liu, Hosni Sassi, Georgina Hernández-Chávez, Jonathan Baert, Laurence Van Melderen, Juan Carlos Fragoso-Jiménez, Guillermo Gosset, Frédéric Goormaghtigh, Paul Gaytán
Jazyk: angličtina
Rok vydání: 2019
Předmět:
0106 biological sciences
Green Fluorescent Proteins
Mutant
Biotechnologie
Glucose Transport Proteins
Facilitative
lcsh:QR1-502
Bioengineering
Acetates
medicine.disease_cause
01 natural sciences
Applied Microbiology and Biotechnology
lcsh:Microbiology
Green fluorescent protein
03 medical and health sciences
chemistry.chemical_compound
Bioreactors
010608 biotechnology
Escherichia coli
medicine
Single cell
Flow cytometry
030304 developmental biology
2. Zero hunger
0303 health sciences
Expression vector
Escherichia coli Proteins
Research
Glucose transporter
Wild type
Biological Transport
PEP group translocation
Microfluidic Analytical Techniques
3. Good health
Kinetics
Glucose
chemistry
Biochemistry
Filamentation
Galactose
Instrumentation médicale
Microbiologie et protistologie [bacteriol.virolog.mycolog.]
Microfluidic imaging
Metabolic engineering
Biotechnology
Zdroj: Microbial Cell Factories, Vol 18, Iss 1, Pp 1-16 (2019)
Microbial Cell Factories
Microbial cell factories, 18 (1
ISSN: 1475-2859
DOI: 10.1186/s12934-019-1073-5
Popis: Background: Escherichia coli W3110 and a group of six isogenic derivatives, each displaying distinct specific rates of glucose consumption were characterized to determine levels of GFP production and population heterogeneity. These strains have single or combinatory deletions in genes encoding phosphoenolpyruvate:sugar phosphotransferase system (PTS) permeases as PtsG and ManX, as well as common components EI, Hpr protein and EIIA, also the non-PTS Mgl galactose/glucose ABC transporter. They have been transformed for expressing GFP based on a lac-based expression vector, which is subject to bistability. Results: These strains displayed specific glucose consumption and growth rates ranging from 1.75 to 0.45 g/g h and 0.54 to 0.16 h-1, respectively. The rate of acetate production was strongly reduced in all mutant strains when compared with W3110/pV21. In bioreactor cultures, wild type W3110/pV21 produced 50.51 mg/L GFP, whereas strains WG/pV21 with inactive PTS IICBGlc and WGM/pV21 with the additional inactivation of PTS IIABMan showed the highest titers of GFP, corresponding to 342 and 438 mg/L, respectively. Moreover, we showed experimentally that bistable expression systems, as lac-based ones, induce strong phenotypic segregation among microbial populations. Conclusions: We have demonstrated that reduction on glucose consumption rate in E. coli leads to an improvement of GFP production. Furthermore, from the perspective of phenotypic heterogeneity, we observed in this case that heterogeneous systems are also the ones leading to the highest performance. This observation suggests reconsidering the generally accepted proposition stating that phenotypic heterogeneity is generally unwanted in bioprocess applications.[Figure not available: see fulltext.]
SCOPUS: ar.j
info:eu-repo/semantics/published
Databáze: OpenAIRE
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