Detection of polyhydroxyalkanoate-accumulating bacteria from domestic wastewater treatment plant using highly sensitive PCR primers.
Polyhydroxyalkanoate (PHA) is a class of biodegradable plastics that have great potential applications in the near future. In this study, the micro-biodiversity and productivity of PHA-accumulating bacteria in activated sludge from a domestic wastewater treatment plant were investigated. A previously reported primer set and a selfdesigned primer set (phaCF1BO/phaCR2BO) were both used to amplify the PHA synthase (phaC) gene of isolated colonies. The new primers demonstrated higher sensitivity for phaC, and combining the PCR results of the two primer sets was able to widen the range of detected genera and raise the sensitivity to nearly 90%. Results showed that 85.3% of the identified bacteria were Gram-negative, with Ralstonia as the dominant genus, and 14.7% were Gram-positive. In addition, Zoogloea and Rhizobium contained the highest amounts of intracellular PHA. It is apparent that glucose was a better carbon source than pentone or tryptone for promoting PHA production in Micrococcus. Two different classes, class I and class II, of phaC were detected from alphaproteobacteria, betaproteobacteria, and gammaproteobacteria, indicating the wide diversity of PHA-accumulating bacteria in this particular sampling site. Simultaneous wastewater treatment and PHA production is promising by adopting the high PHAaccumulating bacteria isolated from activated sludge.
Department of Bioenvironmental Engineering and R&D Center for Membrane Technology, Chung Yuan Christian University, Chung-Li 32023, Taiwan. yt_huang@cycu.edu.tw
Yu-Tzu
YT
Huang
Pi-Ling
PL
Chen
Galilee Uy
GU
Semblante
Sheng-Jie
SJ
You
D000217:Acyltransferases / Q000235:genetics; D001419:Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001431:Bacteriological Techniques / Q000379:methods*; D044822:Biodiversity; D017931:DNA Primers / Q000235:genetics; D054813:Polyhydroxyalkanoates / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012722:Sewage / Q000382:microbiology*
D017931:DNA Primers; D054813:Polyhydroxyalkanoates; D012722:Sewage; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase