title: A novel nucleic lateral flow assay for screening of PHA-producing haloarchaea.
abstract: Polyhydroxyalkanoates (PHAs) are important for biodegradable plastic production, and prokaryotes play a very important role in PHA production. PHA synthase is a key enzyme for the polymerization of PHAs. There are four classes of PHA synthase. The phaC gene is necessary for the production of all classes of PHA synthase, whereas the phaE gene is necessary for the production of class III PHA synthase. This gene is a biomarker for microorganisms that contain class III PHA synthase, such as haloarchaea. Standard techniques for screening of PHA-producing haloarchaea require time for culturing and have poor specificity and sensitivity. Thus, the phaE biosensor was developed to overcome these issues. PCR and DNA lateral flow biosensor techniques were combined for construction of the phaE biosensor. The phaE biosensor has a high specificity for PHA-producing haloarchaea. The lowest amount of genomic DNA of Haloquadratum walsbyi DSM 16854 that the phaE gene could be detected by the biosensor was approximately 250 fg. The phaE biosensor can be applied for screening of PHA-producing haloarchaea from environmental samples. The phaE biosensor is easy to handle and dispose. For screening PHA-producing haloarchaea, the phaE biosensor requires less time and costs less than the standard methods.
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Rd., Rajthevi, Bangkok 10400, Thailand.
authors.forename: Wannaporn
authors.initials: W
authors.lastname: Muangsuwan
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Rd., Rajthevi, Bangkok 10400, Thailand.
authors.forename: Pattarawan
authors.initials: P
authors.lastname: Ruangsuj
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Rd., Rajthevi, Bangkok 10400, Thailand.
authors.forename: Pichai
authors.initials: P
authors.lastname: Chaichanachaicharn
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Rd., Rajthevi, Bangkok 10400, Thailand. Electronic address: montri.yas@mahidol.ac.th.
authors.forename: Montri
authors.initials: M
authors.lastname: Yasawong
mesh_terms: D000217:Acyltransferases / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D015374:Biosensing Techniques / Q000191:economics / Q000379:methods*; D003001:Cloning, Molecular; D019605:Euryarchaeota / Q000201:enzymology* / Q000235:genetics*; D005798:Genes, Bacterial; D054813:Polyhydroxyalkanoates / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D054813:Polyhydroxyalkanoates; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase
keywords: Biosensor; DNA lateral flow; Halophiles; PHA synthase class III; PHAs; phaE