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Development of Klebsiella pneumoniae J2B as microbial cell factory for the production of 1,3-propanediol from glucose
Development of Klebsiella pneumoniae J2B as microbial cell factory for the production of 1,3-propanediol from glucose
- Source :
- Metabolic Engineering. 62:116-125
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- 1,3-Propanediol (1,3-PDO) is an important platform chemical which has a wide application in food, cosmetics, pharmaceutical and textile industries. Its biological production using recombinant Escherichia coli with glucose as carbon source has been commercialized by DuPont, but E. coli cannot synthesize coenzyme B12 which is an essential and expensive cofactor of glycerol dehydratase, a core enzyme in 1,3-PDO biosynthesis. This study aims to develop a more economical microbial cell factory using Klebsiella pneumoniae J2B which can naturally synthesize coenzyme B12. To this end, the heterologous pathway for the production of glycerol from dihydroxyacetone-3-phosphate (DHAP), a glycolytic intermediate, was introduced to J2B and, afterwards, the strain was extensively modified for carbon and energy metabolisms including: (i) removal of carbon catabolite repression, (ii) blockage of glycerol export across the cell membrane, (iii) improvement of NADH regeneration/availability, (iv) modification of TCA cycle and electron transport chain, (v) overexpression of 1,3-PDO module enzyme, and (vi) overexpression of glucose transporter. A total of 33 genes were modified and/or overexpressed, and one resulting strain could produce 814 mM (62 g/L) of 1,3-PDO with the yield of 1.27 mol/mol glucose in fed-batch bioreactor culture with a limited supplementation of coenzyme B12 at 4 μM, which is ~10 fold less than that employed by DuPont. This study highlights the importance of balanced use of glucose in the production of carbon backbone of the target chemical (1,3-PDO) and regeneration of reducing power (NADH). This study also suggests that K. pneumoniae J2B is a promising host for the production of 1,3-PDO from glucose.
- Subjects :
- Glycerol
0106 biological sciences
Glycerol dehydratase
Catabolite repression
NADH regeneration
Bioengineering
01 natural sciences
Applied Microbiology and Biotechnology
Cofactor
03 medical and health sciences
chemistry.chemical_compound
010608 biotechnology
Escherichia coli
1,3-Propanediol
030304 developmental biology
0303 health sciences
biology
Glucose transporter
Citric acid cycle
Klebsiella pneumoniae
Glucose
chemistry
Biochemistry
Propylene Glycols
biology.protein
Biotechnology
Subjects
Details
- ISSN :
- 10967176
- Volume :
- 62
- Database :
- OpenAIRE
- Journal :
- Metabolic Engineering
- Accession number :
- edsair.doi.dedup.....f0e216e285af068bf904be2764b4de8e
- Full Text :
- https://doi.org/10.1016/j.ymben.2020.09.001