Bio-based production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with modulated monomeric fraction in Escherichia coli

In this study, we applied metabolic engineering and bioprocessing strategies to enhance heterologous production of an important biodegradable copolymer, i.e., poly(3-hydroxybutyrate- co -3-hydroxyvalerate) (PHBV), with a modulated 3-hydroxyvalerate (3-HV) monomeric fraction from structurally unrelat...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2021-02, Vol.105 (4), p.1435-1446
Main Authors: Miscevic, Dragan, Mao, Ju-Yi, Mozell, Bradley, Srirangan, Kajan, Abedi, Daryoush, Moo-Young, Murray, Chou, C. Perry
Format: Article
Language:English
Subjects:
Biodegradability
Biodegradation
Biomedical and Life Sciences
Bioprocessing
Biosynthesis
Biotechnological Products and Process Engineering
Biotechnology
Carbon
Cell metabolism
Channeling
Copolymers
Cultivation
E coli
Escherichia coli
Escherichia coli - genetics
Genetic aspects
Glycerol
Hydroxybutyrates
Life Sciences
Mechanical properties
Metabolic engineering
Microbial Genetics and Genomics
Microbiology
Pentanoic Acids
Physiological aspects
Polyesters
Propanol
Succinyl-CoA
Tricarboxylic acid cycle
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Summary:In this study, we applied metabolic engineering and bioprocessing strategies to enhance heterologous production of an important biodegradable copolymer, i.e., poly(3-hydroxybutyrate- co -3-hydroxyvalerate) (PHBV), with a modulated 3-hydroxyvalerate (3-HV) monomeric fraction from structurally unrelated carbon of glycerol in engineered Escherichia coli under different oxygenic conditions. We used our previously derived propanologenic (i.e., 1-propanol-producing) E. coli strain with an activated genomic Sleeping beauty mutase (Sbm) operon as a host for heterologous expression of the phaCAB operon. The 3-HV monomeric fraction was modulated by regulating dissimilated carbon flux channeling from the tricarboxylic acid (TCA) cycle into the Sbm pathway for biosynthesis of propionyl-CoA, which is a key precursor to ( R )-3-hydroxyvaleryl-CoA (3-HV-CoA) monomer. The carbon flux channeling was regulated either by manipulating a selection of genes involved in the TCA cycle or varying oxygenic condition of the bacterial culture. With these consolidated strategies being implemented, we successfully achieved high-level PHBV biosynthesis with a wide range of 3-HV monomeric fraction from ~ 4 to 50 mol%, potentially enabling the fine-tuning of PHBV mechanical properties at the biosynthesis stage. We envision that similar strategies can be applied to enhance bio-based production of chemicals derived from succinyl-CoA. Key points • TCA cycle engineering was applied to enhance 3-HV monomeric fraction in E. coli. • Effects of oxygenic conditions on 3-HV incorporation into PHBV in E. coli were investigated. • Bacterial cultivation for high-level PHBV production in engineered E. coli was performed.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-021-11108-1