Specific identification of (R)-3-hydroxyacyl-ACP: CoA transacylase gene from Pseudomonas and Burkholderia strains by polymerase chain reaction.
Polyhydroxyalkanoates (PHA) were biodegradable thermoplastics. Due to their broad applications, direct biosynthesis of PHA from inexpensive substrates, such as carbohydrates, is actively pursued. It has been recently revealed that (R)-3-hydroxyacyl-ACP: CoA transacylase (PhaG) played an important role in this pathway. In this study, a polymerase chain reaction (PCR) protocol was developed for the rapid and specific identification of phaG gene from various bacteria. Using the PCR strategy, the complete open reading frames of two phaG genes from Pseudomonas stutzeri 1317 and Pseudomonas nitroreducens 0802 were cloned from the genomic DNA and functionally expressed in Pseudomonas putida PHAGN-21. Furthermore, this strategy was successful applied in non-Pseudomonas strains, such as Burkholderia. These results suggest that PhaG-mediated pathway of medium-chain-length polyhydroxyalkanoates was widespread among bacteria.
Zheng
Zhong
Z
MOE Laortoy of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua Unieirsty, Beiing 100084, China.
Chen
Jin-Chun
JC
Tian
Hong-Lei
HL
Bei
Feng-Feng
FF
Chen
Guo-Qiang
GQ
D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D019117:Burkholderia / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D054813:Polyhydroxyalkanoates / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment
D054813:Polyhydroxyalkanoates; D000217:Acyltransferases; C114682:3-hydroxyacyl-CoA-acyl carrier protein transferase