Abstract: Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.
Abstract: Approximately 10,000 different types of dyes and
pigments are being used in various industrial applications yearly,
which include the textile and printing industries. However, these dyes
are difficult to degrade naturally once they enter the aquatic system.
Their high persistency in natural environment poses a potential health
hazard to all form of life. Hence, there is a need for alternative dye
removal strategy in the environment via bioremediation. In this study,
fungi laccase is investigated via commercial agar dyes plates and
submerged fermentation to explore the application of fungi laccase in
textile dye wastewater treatment. Two locally isolated basidiomycetes
were screened for laccase activity using media added with commercial
dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid
(ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3
(1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS
plates with the appearance of greenish halo on around the isolates.
Submerged fermentation performed on Isolate TBB3 with the
productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate
EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed
higher laccase production, it was subjected to molecular
characterization by DNA isolation, PCR amplification and sequencing
of ITS region of nuclear ribosomal DNA. After being compared with
other sequences in National Center for Biotechnology Information
(NCBI database), isolate TBB3 is probably from species Trametes
hirsutei. Further research work can be performed on this isolate by
upscale the production of laccase in order to meet the demands of the
requirement for higher enzyme titer for the bioremediation of textile
dyes.
Abstract: Extraction of laccase produced by L. polychrous in an
aqueous two-phase system, composed of polyethylene glycol and
phosphate salt at pH 7.0 and 250C was investigated. The effect of
PEG molecular weight, PEG concentration and phosphate
concentration was determined. Laccase preferentially partitioned to
the top phase. Good extraction of laccase to the top phase was
observed with PEG 4000. The optimum system was found in the
system containing 12% w/w PEG 4000 and 16% w/w phosphate salt
with KE of 88.3, purification factor of 3.0-fold and 99.1% yield.
Some properties of the enzyme such as thermal stability, effect of
heavy metal ions and kinetic constants were also presented in this
work. The thermal stability decreased sharply with high temperature
above 60 0C. The enzyme was inhibited by Cd2+, Pb2+, Zn2+ and
Cu2+. The Vmax and Km values of the enzyme were 74.70
μmol/min/ml and 9.066 mM respectively.