1,3-Propanediol production by Escherichia coli using genes from Citrobacter freundii atcc 8090.
Compared with chemical synthesis, fermentation has the advantage of mass production at low cost, and has been used in the production of various industrial chemicals. As a valuable organic compound, 1,3-propanediol (1,3-PDO) has numerous applications in the production of polymers, lubricants, cosmetics and medicines. Here, conversion of glycerol (a renewable substrate and waste from biodiesel production) to 1,3-PDO by E. coli bacterial strain carrying altered glycerol metabolic pathway was investigated. Two gene constructs containing the 1,3-PDO operon from Citrobacter freundii (pCF1 and pCF2) were used to transform the bacteria. The pCF1 gene expression construct contained dhaBCE genes encoding the three subunits of glycerol dehydratase, dhaF encoding the large subunit of the glycerol dehydratase reactivation factor and dhaG encoding the small subunit of the glycerol dehydratase reactivating factor. The pCF2 gene expression construct contained the dhaT gene encoding the 1,3-propanediol dehydrogenase. Expression of the genes cloned in the above constructs was under regulation of the T7lac promoter. RT-PCR, SDS-PAGE analyses and functional tests confirmed that 1,3-PDO synthesis pathway genes were expressed at the RNA and protein levels, and worked flawlessly in the heterologous host. In a batch flask culture, in a short time applied just to identify the 1,3-PDO in a preliminary study, the recombinant E. coli bacteria produced 1.53 g/L of 1,3-PDO, using 21.2 g/L of glycerol in 72 h. In the Sartorius Biostat B Plus reactor, they produced 11.7 g/L of 1,3-PDO using 24.2 g/L of glycerol, attaining an efficiency of 0.58 [mol1,3-PDO/molglycerol].
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland.
Hanna
H
Przystałowska
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland.
Joanna
J
Zeyland
Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, Poznań, Poland.
Alicja
A
Kośmider
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland; Polish Academy of Sciences, Institute of Human Genetics, Poznań, Poland.
Marlena
M
Szalata
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland; Polish Academy of Sciences, Institute of Human Genetics, Poznań, Poland.
Ryszard
R
Słomski
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland; Polish Academy of Sciences, Institute of Human Genetics, Poznań, Poland.
Daniel
D
Lipiński
D000251:Adenosine Triphosphatases / Q000235:genetics; D000426:Alcohol Dehydrogenase / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D019149:Bioreactors; D002851:Chromatography, High Pressure Liquid; D016971:Citrobacter freundii / Q000235:genetics* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005285:Fermentation; D005990:Glycerol / Q000737:chemistry; D006836:Hydro-Lyases / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D011401:Promoter Regions, Genetic; D011409:Propylene Glycols / Q000737:chemistry*; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature
D001426:Bacterial Proteins; D011108:Polymers; D011409:Propylene Glycols; D011994:Recombinant Proteins; C041787:1,3-propanediol; D012313:RNA; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase; D000251:Adenosine Triphosphatases; C426653:DhaG protein, Citrobacter freundii; D006836:Hydro-Lyases; C020671:glycerol dehydratase; D005990:Glycerol