Recovery of laccase-producing gammaproteobacteria from wastewater

Wastewater environment is a rich source of microorganisms with the capability for the degradation of malicious aromatic pollutants. Although wastewater could be regarded as both a resource and a problem, we intended to elucidate its beneficial aspect in this study sourcing for laccase-producing proteobacteria. Different wastewater samples, from selected wastewater treatment plants (WWTPs), were selectively enriched with some model compounds (vanillin, lignin and potassium hydrogen phthalate) to screen out bacterial isolates that possess excellent degradation potentials. Thereafter, positive isolates were screened for the production of laccase and degradation on phenolic (guaiacol, α-naphthol and syringaldazine) and non-phenolic (ABTS; 2,2 azino-bis -(3-ethylbenzothiazoline 6 sulphonic acid) and PFC; potassium ferrocyanoferrate) substrates characteristic of laccase oxidation. Remarkable laccase producers were identified based on their 16 S rRNA sequences and their secreted enzymes were subjected to substrate specificity test, employing laccase substrates; ABTS, PFC, guaiacol, α-naphthol, 2,6-dimethoxyphenol and pyrogallol. Results showed that wastewater and selective enrichment, in tandem, produced the gammaproteobacteria Pseudomonas aeruginosa DEJ16, Pseudomonas mendocina AEN16 and Stenotrophomonas maltophila BIJ16, which preferably oxidized the non-phenolic substrates. Units of extracellular laccase activity ranging between cca. 490 and cca. 600 U/mL were recorded for ABTS whereas outputs recorded from PFC catalysis ranged from cca. 320 to cca. 430 U/mL. Stenotrophomonas maltophila BIJ16 presented an unparalleled high laccase activity and had a responsive substrate specificity to aromatic and inorganic substrates, thereby suggesting its employment for in situ biodegradation studies. In conclusion, wastewater serves as an ideal milieu for the isolation of laccase producing bacteria.