Copper induction and differential expression of laccase in Aspergillus flavus

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.

Constitutive and inducible pectinolytic enzymes from Aspergillus flavipes FP-500 and their modulation by pH and carbon source / Enzimas pectinolíticas constitutivas e indutíveis de Aspergillus flavipes FP-500 e sua modulação pelo pH e fonte de carbono

Growth and enzymes production by Aspergillus flavipes FP-500 were evaluated on pectin, polygalacturonic acid, galacturonic acid, arabinose, rhamnose, xylose, glycerol and glucose at different initial pH values. We found that the strain produced exopectinases, endopectinases and pectin lyases. Exopectinases and pectin lyase were found to be produced at basal levels as constitutive enzymes and their production was modulated by the available carbon source and pH of culture medium and stimulated by the presence of inducer in the culture medium. Endo-pectinase was basically inducible and was only produced when pectin was used as carbon source. Our results suggest that pectinases in A. flavipes FP-500 are produced in a concerted way. The first enzyme to be produced was exopectinase followed by Pectin Lyase and Endo-pectinase.

Avaliou-se o crescimento e a produção de enzimas por Aspergillus flavipes FP-500 em pectina, ácido poligalacturônico, ácido galacturônico, arabinose, ramnose, xilose, glicerol e glicose, em diferentes valores de pH inicial. Verificamos que a cepa produziu exopectinases, endopectinases e pectina liases. Exopectinases e pectina liases foram produzidas em níveis basais como enzimas constitutivas e sua produção foi modulada pela fonte de carbono disponível e pelo pH do meio de cultura e estimulada pela presença de indutores no meio de cultura. Endopectinase foi indutível e produzida somente quando pectina foi utilizada como fonte de carbono. Nossos resultados sugerem que as pectinases de A. flavipes FP-500 são produzidas de forma planejada. A primeira enzima a ser produzida foi expopectinase, seguida por pectina liase e endopectinase.

Enzymatic characteristics of quercetinases from some indigenous Aspergillus flavus strains.

Five indigenous Aspergillus flavus strains MTCC-2206, 1884, 1883, 1783 and 2456 were screened for the secretion of quercetinase. Fungal strains MTCC-2206, 1884, 1883, and 1783 were found to secrete the quercetinase in the range of 0.24-0.36 enzyme unit/mL of the culture medium, while MTCC-2456 secreted only 0.04 enzyme unit/mL. The enzymatic characteristics of quercetinase were determined. The Km values using quercetin as the substrate were 12.5 microM, 14.0 microM, 12.5 microM and 13.0 microM for the quercetinase produced by MTCC-2206, 1884, 1883 and 1783, respectively. The pH optima for the above enzymes were 6.5, 6.5, 6.0 and 6.0 and temperature optima were 45, 40, 45 and 50 degrees C, respectively. The partial purification from only one strain MTCC-2206 was achieved (nearly 3-fold purification).