Optimization of laccase production from Aspergillus flavus by design of experiment technique: Partial purification and characterization

Abstract

The present study describes isolation of laccase producing fungal strain and optimization of the process parameters by design of experiment (DOE) technique to achieve the maximum production of extracellular laccases by Aspergillus flavus obtained from natural habitat. Bromophenol blue dye and ABTS (2,2 0 -azinobis 3-ethyl-benzothiazoline-6-sulfonate) were used as substrates for the screening of laccase activity. Design expert 8.0.7.1 software was used to optimize culture conditions such as carbon source, nitrogen source, temperature and pH. Subsequently, optimization for inoculums size was also carried out. The optimization studies revealed that the laccase yield was highest when operated at the following conditions: carbon source -cellulose (8%), nitrogen source -peptone (2%), temperature -35°C, pH -7 and inoculum of size 1.5 cm.

Isolation, Screening and Optimization for Laccase production by Scytalidium lignicola pesante under submerged fermentation

“…Both the isolates showed characteristic blue color zone surrounding the colonies due to the oxidation of ABTS. ABTS assay is considered to be more efficient assay for screening of laccase producer as far as cost, time and method is concerned (Kumar et al, 2016). None of the two isolates showed zone on the Azure B containing plates.…”

“…None of the two isolates showed zone on the Azure B containing plates. According to Kumar et al, (2016) several fungi which secrete lignin peroxidase and manganese peroxidase have ability to decolorize or degrade Azure B dye.…”

“…To this two discs (8mm in diameter) of a 5 days old culture were inoculated and incubated at ± 25ᵒC for 10 days. The culture harvested every day from day 2 to day 10 and was used to determine enzyme activity (Kumar et al, 2016).…”

Enzim Laccase dari Edible Mushroom untuk Pemutihan Pati Sagu Ramah Lingkungan

“…Kultur fermentasi cair jamur edible A. bisporus dan P. ostratus menggunakan media Potato Dextrose Broth (PDB) mengacu pada metode (10,21) yang dimodifikasi. Media PDB yang diperkaya dengan mikromineral dilarutkan dalam 100 mL air destilasi pada 250 mL flask Erlenmeyer.…”

“…Media PDB yang diperkaya dengan mikromineral dilarutkan dalam 100 mL air destilasi pada 250 mL flask Erlenmeyer. A.bisporus, dan R.oligosporus menggunakan bahan media fermentasi cair mengacu pada metode (10,21) yang dimodifikasi dengan substitusi sumber karbon dan sumber protein. Sampling dilakukan pada hari ke-0, 3, 6, 9, 12, 15, 18, dan 21.…”