Optimisation and physicochemical characterisation of a thermo-alkali stable laccase produced by wastewater associated Bacillus sp. NU2.

Laccase is a multicopper enzyme that plays a unique role in bioremediation of environmental pollutants. Bacteria were isolated from hospital wastewater and screened for laccase production. The laccase production process condition was optimised, and the laccase obtained was characterised. The 16S rRNA molecular analysis conducted on the best laccase producer revealed a Bacillus sp. NU2 identified. The process conditions: pH5, 45°C, 100 rpm, 5% inoculum, and growth constituents viz: tangerine peel and wheat bran agro-wastes, beef extract, ammonium persulfate, glucose, galactose, xylose, sorbitol, fructose carbon sources; and 4-aminophenol inducer optimally stimulated laccase production. The Bacillus sp. NU2 laccase was optimal at pH and temperature conditions of 8.0°C and 60°C, with a noteworthy pH and thermal stability observed. Furthermore, NU2 laccase showed a moderate/high tolerance and relative activity effect on various chemical inhibitors, halides and surfactant of triton x-100 (105 ± 0.92%), PMSF (107 ± 0.81%), and NaCl (94 ± 0.81%) at 1, 3, and 6 (mM) concentration. Additionally, NU2 laccase maintained a relative activity of 101%, 104%, and 102% for Mg2+, Zn2+, and Fe3+ at 1, 3, and 6 mM respectively. Acetone and propanol significantly upregulated laccase activity at 114 ± 0.0008% and 118.24 ± 0.35 and also at 30 and 20 (%) concentrations. Conclusively, the tolerant effect of Bacillus sp. NU2 laccase in pH, temperature, inhibitors and organic solvents suggests its potential for biotechnological application and promotion of a greener environment.

Marine sediment derived bacteria Enterobacter asburiae ES1 and Enterobacter sp. Kamsi produce laccase with high dephenolisation potentials.

Purified laccases from bacterial species isolated from marine sediment were applied to degrade Bisphenol A (BPA). The Bacterial species were isolated from marine water sediments sampled from Cove Rock and Bonza Bay beach of the Eastern Cape Province, South Africa was tested for laccase activity on varied phenolic plates. The two most promising strains, Enterobacter asburiae ES1 and Enterobacter sp. Kamsi was subjected to extracellular laccase production and were identified using molecular methods. Both extracted bacterial laccases showed an affinity for ABTS and PFC substrates and were purified to homogeneity by ammonium sulfate precipitation, anion exchange, and size exclusion chromatography. A specific laccase activity of 231.67 and 218.15 U/mg of protein and a molecular weight of 50 and 55 kDa was obtained from the purified ES1 and Kamsi laccases. Laccase activity was optimum at pH8 and 5 and at 80 °C and 60 °C for ES1 and Kamsi laccases, and they manifested 71.7% and 65.8% BPA decolorizing effects. The optimized treatment condition applied showed maximum BPA removal effects of 85% and 86% at pH7 and 6, while 78% and 79% was degraded at 70 °C and 80 °C while at 250 µL enzyme volume, BPA was actively degraded to 85%, and 75% removal effect showed by ES1 and Kamsi laccases. The molecular identification of the pure colonies using 16S rRNA showed the isolate belonged to the class of gammaproteobacterial. Their nucleotide sequence has been deposited in NCBI with the accession number MN686602 and MN686603. Conclusively, marine habitat serves as a reservoir for active bacterial laccase producers suitable for bioprocess application.