Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from gluconate and glucose by recombinant Aeromonas hydrophila and Pseudomonas putida.
Aeromonas hydrophila 4AK4 and Pseudomonas putida GPp104 were genetically engineered to synthesize poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) using gluconate and glucose rather than fatty acids. A truncated tesA gene, encoding cytosolic thioesterase I of Escherichia coli which catalyzes the conversion of acyl-ACP into free fatty acids, was introduced into A. hydrophila 4AK4. When grown in gluconate, the recombinant A. hydrophila 4AK4 synthesized 10% (w/w) PHBHHx containing 14% (mol/mol) 3-hydroxyhexanoate. If additional PHBHHx synthesis genes, phaPCJ, were over-expressed with the truncated tesA in A. hydrophila 4AK4, the PHBHHx content increased to 15% (w/w) and contained 19% (mol/mol) 3-hydroxyhexanoate. Recombinant P. putida GPp104 harboring phaC encoding PHBHHx synthase of A. hydrophila, phaB encoding acetoacetyl-CoA reductase of Wautersia eutropha and phaG encoding 3-hydroxyacyl-ACP-CoA transferase of P. putida, synthesized 19% (w/w) PHBHHx containing 5% (mol/mol) 3-hydroxyhexanoate from glucose. The results suggest that the engineered pathways were applicable to synthesize PHBHHx from unrelated carbon sources such as gluconate and glucose.
Qiu
Yuan-Zheng
YZ
Department of Biological Sciences and Biotechnology, Tsinghua University, 100084, Beijing, China.
Han
Jing
J
Guo
Jing-Jing
JJ
Chen
Guo-Qiang
GQ
D020155:3-Hydroxybutyric Acid / Q000378:metabolism*; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D016980:Aeromonas hydrophila / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D000429:Alcohol Oxidoreductases; D002208:Caproates / Q000378:metabolism*; D020118:Cupriavidus necator / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods; D005942:Gluconates / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D008245:Lysophospholipase / Q000235:genetics / Q000378:metabolism; D033903:Periplasmic Proteins / Q000235:genetics / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013869:Thiolester Hydrolases / Q000235:genetics / Q000378:metabolism; D000094122:Polyhydroxybutyrates
D020155:3-Hydroxybutyric Acid; D000217:Acyltransferases; D000429:Alcohol Oxidoreductases; D002208:Caproates; D029968:Escherichia coli Proteins; D005942:Gluconates; D005947:Glucose; D008245:Lysophospholipase; D033903:Periplasmic Proteins; D011994:Recombinant Proteins; D013869:Thiolester Hydrolases; C115940:poly(3-hydroxybutyrate-co-3-hydroxyhexanoate); D000094122:Polyhydroxybutyrates; C045051:3-hydroxyhexanoic acid; C023744:acetoacetyl-CoA reductase; C114682:3-hydroxyacyl-CoA-acyl carrier protein transferase; C077225:poly(3-hydroxyalkanoic acid) synthase; C400724:tesA protein, E coli; C030691:gluconic acid