Biosynthesis of polyhydroxyalkanoates from sucrose by metabolically engineered Escherichia coli strains.

Sucrose utilization has been established in Escherichia coli strains by expression of Mannheimia succiniciproducens β-fructofuranosidase (SacC), which hydrolyzes sucrose into glucose and fructose. Recombinant E. coli strains that can utilize sucrose were examined for their abilities to produce poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] from sucrose. When recombinant E. coli strains expressing Ralstonia eutropha PhaCAB and SacC were cultured in MR medium containing 20 g/L of sucrose, all recombinant E. coli strains could produce P(3HB) from sucrose. Also, recombinant E. coli strains expressing Pseudomonas sp. MBEL 6-19 PhaC1437, Clostridium propionicum Pct540, R. eutropha PhaAB enzymes along with SacC could produce P(3HB-co-LA) from sucrose. Among the examined E. coli strains, recombinant E. coli XL1-Blue produced the highest contents of P(3HB) (53.60 ± 2.55 wt%) and P(3HB-co-LA) (29.44 ± 0.39 wt%). In the batch fermentations, recombinant E. coli XL1-Blue strains completely consumed 20 g/L of sucrose as the sole carbon source and supported the production of 3.76 g/L of P(3HB) and 1.82 g/L of P(3HB-co-LA) with 38.21 wt% P(3HB) and 20.88 wt% P(3HB-co-LA) contents, respectively. Recombinant E. coli strains developed in this study can be used to establish a cost-efficient biorefinery for the production of polyhydroxyalkanoates (PHAs) from sucrose, which is an abundant and inexpensive carbon source.

[1]  Guoqiang Chen,et al.  Engineering microorganisms for improving polyhydroxyalkanoate biosynthesis. , 2018, Current opinion in biotechnology.

[2]  Hyun Uk Kim,et al.  Enhanced production of poly‑3‑hydroxybutyrate (PHB) by expression of response regulator DR1558 in recombinant Escherichia coli. , 2019, International journal of biological macromolecules.

[3]  S. Lee,et al.  Metabolic engineering of Escherichia coli for the production of l-valine based on transcriptome analysis and in silico gene knockout simulation , 2007, Proceedings of the National Academy of Sciences.

[4]  S. Lee,et al.  Metabolic engineering of Escherichia coli for the production of polylactic acid and its copolymers , 2010, Biotechnology and bioengineering.

[5]  M. K. Gouda,et al.  Production of PHB by a Bacillus megaterium strain using sugarcane molasses and corn steep liquor as sole carbon and nitrogen sources. , 2001, Microbiological research.

[6]  S. Hong,et al.  Recent advances in development of biomass pretreatment technologies used in biorefinery for the production of bio-based fuels, chemicals and polymers , 2015, Korean Journal of Chemical Engineering.

[7]  이승환,et al.  재조합 대장균에서 수크로즈로부터의 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구 , 2014 .

[8]  Dae-Hee Lee,et al.  Biological Valorization of Poly(ethylene terephthalate) Monomers for Upcycling Waste PET , 2019, ACS Sustainable Chemistry & Engineering.

[9]  Sang Yup Lee,et al.  Metabolic engineering of Ralstonia eutropha for the biosynthesis of 2-hydroxyacid-containing polyhydroxyalkanoates. , 2013, Metabolic engineering.

[10]  Choul‐Gyun Lee,et al.  Lipid Extraction from Tetraselmis sp. Microalgae for Biodiesel Production Using Hexane-based Solvent Mixtures , 2018, Biotechnology and Bioprocess Engineering.

[11]  H. Zhang,et al.  Production of polyhydroxyalkanoates in sucrose-utilizing recombinant Escherichia coli and Klebsiella strains , 1994, Applied and environmental microbiology.

[12]  L. Nielsen,et al.  Molecular Control of Sucrose Utilization in Escherichia coli W, an Efficient Sucrose-Utilizing Strain , 2012, Applied and Environmental Microbiology.

[13]  Hee Taek Kim,et al.  Recent advances in metabolic engineering of Corynebacterium glutamicum as a potential platform microorganism for biorefinery , 2018, Biofuels, Bioproducts and Biorefining.

[14]  H. Park,et al.  Production of cis-Vaccenic Acid-oriented Unsaturated Fatty Acid in Escherichia coli , 2018, Biotechnology and Bioprocess Engineering.

[15]  J. Yang,et al.  Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose. , 2015, Biotechnology and bioengineering.

[16]  S. Lee,et al.  Advanced bacterial polyhydroxyalkanoates: towards a versatile and sustainable platform for unnatural tailor-made polyesters. , 2012, Biotechnology advances.

[17]  Anthony L Andrady,et al.  Microplastics in the marine environment. , 2011, Marine pollution bulletin.

[18]  S. Lee,et al.  Systems Metabolic Engineering Strategies for Non‐Natural Microbial Polyester Production , 2019, Biotechnology journal.

[19]  Chi-Wei Lo,et al.  Isolation and purification of bacterial poly(3-hydroxyalkanoates) , 2008 .

[20]  Sang Yup Lee,et al.  Metabolic engineering for the synthesis of polyesters: A 100-year journey from polyhydroxyalkanoates to non-natural microbial polyesters. , 2020, Metabolic engineering.

[21]  Hee Taek Kim,et al.  Recent Advances in the Metabolic Engineering of Klebsiella pneumoniae: A Potential Platform Microorganism for Biorefineries , 2019, Biotechnology and Bioprocess Engineering.

[22]  Hee Taek Kim,et al.  Metabolic engineering of Corynebacterium glutamicum for fermentative production of chemicals in biorefinery , 2018, Applied Microbiology and Biotechnology.

[23]  Jin Yin,et al.  Engineering NADH/NAD+ ratio in Halomonas bluephagenesis for enhanced production of polyhydroxyalkanoates (PHA). , 2018, Metabolic engineering.

[24]  Tong Un Chae,et al.  Metabolic engineering of Escherichia coli for enhanced biosynthesis of poly(3-hydroxybutyrate) based on proteome analysis , 2013, Biotechnology Letters.

[25]  A. Steinbüchel,et al.  Ralstonia eutropha Strain H16 as Model Organism for PHA Metabolism and for Biotechnological Production of Technically Interesting Biopolymers , 2008, Journal of Molecular Microbiology and Biotechnology.

[26]  M. A. Prieto,et al.  The role of GlpR repressor in Pseudomonas putida KT2440 growth and PHA production from glycerol. , 2013, Environmental microbiology.

[27]  Jeong Wook Lee,et al.  Development of sucrose-utilizing Escherichia coli K-12 strain by cloning β-fructofuranosidases and its application for l-threonine production , 2010, Applied Microbiology and Biotechnology.

[28]  Sang Yup Lee,et al.  Systems metabolic engineering for chemicals and materials. , 2011, Trends in biotechnology.

[29]  Mee-Jung Han,et al.  Proteome Analysis of Metabolically EngineeredEscherichia coli Producing Poly(3-Hydroxybutyrate) , 2000, Journal of bacteriology.

[30]  Guoqiang Chen,et al.  Engineering biosynthesis of polyhydroxyalkanoates (PHA) for diversity and cost reduction. , 2020, Metabolic engineering.

[31]  Jay D. Keasling,et al.  Technical Advances to Accelerate Modular Type I Polyketide Synthase Engineering towards a Retro-biosynthetic Platform , 2019, Biotechnology and Bioprocess Engineering.

[32]  S. Lee,et al.  Biosynthesis of polylactic acid and its copolymers using evolved propionate CoA transferase and PHA synthase , 2010, Biotechnology and bioengineering.

[33]  S. Lee,et al.  Comparison of recombinant Escherichia coli strains for synthesis and accumulation of poly‐(3‐hydroxybutyric acid) and morphological changes , 1994, Biotechnology and bioengineering.

[34]  R. M. Lafferty,et al.  A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass , 1978, European journal of applied microbiology and biotechnology.

[35]  Jae Hyeon Lee,et al.  Production of 1,3-Propanediol from Glucose by Recombinant Escherichia coli BL21(DE3) , 2018, Biotechnology and Bioprocess Engineering.

[36]  Hee Taek Kim,et al.  Development of Metabolically Engineered Corynebacterium glutamicum for Enhanced Production of Cadaverine and Its Use for the Synthesis of Bio-Polyamide 510 , 2020, ACS Sustainable Chemistry & Engineering.