Optimization of laccase production from Trametes versicolor by solid fermentation

2007 ◽  
Vol 53 (2) ◽  
pp. 245-251 ◽  
Author(s):  
Debing Jing ◽  
Peijun Li ◽  
Frank Stagnitti ◽  
Xianzhe Xiong
Keyword(s):  
Trametes Versicolor ◽  
Laccase Activity ◽  
Uniform Design ◽  
Tween 80 ◽  
Multivariate Regression Analysis ◽  
Dry Mass ◽  
Solid Substrate ◽  
Culture Conditions ◽  
Laccase Production ◽  
Solid Fermentation

The regulation of culture conditions, especially the optimization of substrate constituents, is crucial for laccase production by solid fermentation. To develop an inexpensive optimized substrate formulation to produce high-activity laccase, a uniform design formulation experiment was devised. The solid fermentation of Trametes versicolor was performed with natural aeration, natural substrate pH (about 6.5), environmental humidity of 60% and two different temperature stages (at 37 °C for 3 days, and then at 30 °C for the next 17 days). From the experiment, a regression equation for laccase activity, in the form of a second-degree polynomial model, was constructed using multivariate regression analysis and solved with unconstrained optimization programming. The optimized substrate formulation for laccase production was then calculated. Tween 80 was found to have a negative effect on laccase production in solid fermentation; the optimized solid substrate formulation was 10.8% glucose, 27.7% wheat bran, 9.0% (NH4)2SO4, and 52.5% water. In a scaled-up verification of solid fermentation at a 10 kg scale, laccase activity from T. versicolor in the optimized substrate formulation reached 110.9 IU/g of dry mass.

scholarly journals Comparative study on laccase activity of white rot fungi under submerged fermentation with different lignocellulosic wastes

BioResources ◽  
2020 ◽  
Vol 15 (4) ◽  
pp. 9166-9179
Author(s):  
Qi An ◽  
Jie Qiao ◽  
Lu-Sen Bian ◽  
Mei-Ling Han ◽  
Xun-You Yan ◽  
...  
Keyword(s):  
Submerged Fermentation ◽  
Laccase Activity ◽  
White Rot Fungi ◽  
Industrial Applications ◽  
Culture Conditions ◽  
White Rot ◽  
Laccase Production ◽  
Poplar Wood ◽  
Lignocellulosic Wastes ◽  
Cottonseed Hull

Different Pleurotus ostreatus and Flammulina velutipes species were compared relative to their ability to produce laccase in submerged fermentation of various lignocellulosic wastes. Fungi cultivation in identical culture conditions revealed wide differences among both species and strains of the same species. The laccase secretion ability of P. ostreatus strains was superior to F. velutipes strains. Maximum laccase production on cottonseed hull, corncob, and poplar wood was secreted by P. ostreatus CY 568, P. ostreatus CCEF 89, and P. ostreatus CY 568, respectively. The nature of lignocellulosic materials played an important role in determining the expression of laccase potential of fungi. The presence of cottonseed hull improved laccase activity and accelerated the rate of enzyme production. Maximum laccase production on cottonseed hull was nearly 1.29-fold and 1.53-fold higher than that on corncob and poplar wood, respectively. Laccase activity was detected in almost all tested strains on cottonseed hull on the first day, while only a few strains on poplar wood and corncob were detected on the first day. These findings will be helpful for selecting the appropriate strain in industrial applications and for optimization of integrated industrial laccase production.

OPTIMIZATION OF LACCASE PRODUCTION BY CURVULARIA LUNATA USING MAIZE COB AS SUBSTRATE

2021 ◽  
Vol 4 (4) ◽  
pp. 460-468
Author(s):  
A. Bello ◽  
D. A. Machido ◽  
A. I. Mohammed-Dabo ◽  
S. A. Ado
Keyword(s):  
Laccase Activity ◽  
Incubation Temperature ◽  
Agricultural Research ◽  
Culture Conditions ◽  
Inoculum Size ◽  
Laccase Production ◽  
Curvularia Lunata ◽  
Processing Unit ◽  
Crude Fibre ◽  
Maize Cob

Researchers are showing interest in laccase because it is able to degrade several lignocellulosic biomass. It is important to optimize laccase production to achieve higher yield at a lower cost using agricultural wastes. This study was aimed at optimizing the culture conditions for laccase production. Previously isolated and characterized laccase producing Curvularia lunata was collected from the laboratory of Microbiology department, ABU, Zaria. Maize cobs which were used as substrate were collected from Seed Processing Unit of Institute for Agricultural Research, Zaria and the proximate composition of the substrate was determined according to AOAC guideline. The optimum fermentation type for laccase production was determined after which the culture conditions for laccase production were optimized. Laccase activity was determination by guaiacol assay. The proximate compositions of the maize cob were found to be Moisture content (3.18%), Crude protein (2.50%), Crude fat (32.20%), Crude fibre (3.15), Ash content (1.88%) and Carbohydrate content (57.09%). Higher laccase activity was recorded under solid state fermentation compared to submerged fermentation. The optimum culture conditions were found to be inoculum size 3 x 5 mm, pH 5, incubation temperature 30 oC and incubation period of 6 days. The culture conditions for laccase production by Curvularia lunata using maize cob as substrate were optimized

scholarly journals The influence of Tween 80 on laccase production by Pleurotus sajor-caju and the efficiency of crude enzyme broth in the removal of bisphenol-A

2018 ◽  
Vol 85 (0) ◽  
Author(s):  
Thaís Silva Teodoro ◽  
Fernanda de Oliveira ◽  
Caroline Poffo ◽  
Laís Pacenko Braga ◽  
Auriciane Arbigaus ◽  
...  
Keyword(s):  
Bisphenol A ◽  
Culture Medium ◽  
Laccase Activity ◽  
C:N Ratio ◽  
Tween 80 ◽  
Crude Enzyme ◽  
Production Costs ◽  
Oxidative Enzymes ◽  
Laccase Production ◽  
Pleurotus Sajor Caju

ABSTRACT: Bisphenol-A is currently considered an environmental pollutant, capable of interfering in the endocrine system of organisms and causing alterations in its development and reproductive system. An alternative method to the chemical treatment of this pollutant has been the use of oxidative enzymes, especially laccases produced by fungi. In order to reduce production costs, agro-industrial waste can be used in the culture medium composition. Nonionic surfactants, which are only slightly toxic to biological membranes, can be applied, as well as Tween 80, to facilitate the excretion of these enzymes into the culture medium. The objectives of this work were: a) characterize the immersion water of banana straw used in the formulation of the culture medium; b) evaluate laccase production by Pleurotus sajor-caju in culture medium with and without addition of Tween 80, through shaken flasks; c) evaluate the efficiency of the crude enzyme broth in degrading bisphenol-A. The shaken flasks were incubated at 30°C for 12 days. The immersion water had a C:N ratio of 13.8, ash percentage of 28.6%, and pH close to neutrality. The addition of Tween 80 on the culture medium (7.5%, m/v) yielded laccase activity and productivity values equal to 3,016.47 U L-1 and 502.7 U L-1 day-1, respectively. These values were 50 and 33.5 times higher than those obtained in the culture medium without addition of Tween 80 for laccase activity and productivity, respectively. The crude enzyme broth degraded 100% of bisphenol-A after 48 hours, regardless of concentration (500, 750 and 1,000 mg L-1).

scholarly journals Chitosan multiple addition enhances laccase production from Trametes versicolor

2015 ◽  
Vol 38 (10) ◽  
pp. 1973-1981 ◽  
Author(s):  
Abiodun Emmanuel Adekunle ◽  
Feng Wang ◽  
Jianhua Hu ◽  
Anzhou Ma ◽  
Chen Guo ◽  
...  
Keyword(s):  
Trametes Versicolor ◽  
Laccase Production ◽  
Multiple Addition

scholarly journals Protein Engineering Approaches to Enhance Fungal Laccase Production in S. cerevisiae

2021 ◽  
Vol 22 (3) ◽  
pp. 1157
Author(s):  
Pablo Aza ◽  
Felipe de Salas ◽  
Gonzalo Molpeceres ◽  
David Rodríguez-Escribano ◽  
Iñigo de la Fuente ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Directed Evolution ◽  
Signal Peptide ◽  
Laccase Activity ◽  
Laccase Production ◽  
Wide Range ◽  
Mating Factor ◽  
Conserved Amino Acids ◽  
Basidiomycete Fungi

Laccases secreted by saprotrophic basidiomycete fungi are versatile biocatalysts able to oxidize a wide range of aromatic compounds using oxygen as the sole requirement. Saccharomyces cerevisiae is a preferred host for engineering fungal laccases. To assist the difficult secretion of active enzymes by yeast, the native signal peptide is usually replaced by the preproleader of S. cerevisiae alfa mating factor (MFα1). However, in most cases, only basal enzyme levels are obtained. During directed evolution in S. cerevisiae of laccases fused to the α-factor preproleader, we demonstrated that mutations accumulated in the signal peptide notably raised enzyme secretion. Here we describe different protein engineering approaches carried out to enhance the laccase activity detected in the liquid extracts of S. cerevisiae cultures. We demonstrate the improved secretion of native and engineered laccases by using the fittest mutated α-factor preproleader obtained through successive laccase evolution campaigns in our lab. Special attention is also paid to the role of protein N-glycosylation in laccase production and properties, and to the introduction of conserved amino acids through consensus design enabling the expression of certain laccases otherwise not produced by the yeast. Finally, we revise the contribution of mutations accumulated in laccase coding sequence (CDS) during previous directed evolution campaigns that facilitate enzyme production.

scholarly journals Production, Survival, and Evaluation of Solid-Substrate Inocula of Penicillium oxalicum, a Biocontrol Agent Against Fusarium Wilt of Tomato

2002 ◽  
Vol 92 (8) ◽  
pp. 863-869 ◽  
Author(s):  
I. Larena ◽  
P. Melgarejo ◽  
A. De Cal
Keyword(s):  
Fusarium Wilt ◽  
Biocontrol Agent ◽  
Solid Substrate ◽  
Penicillium Oxalicum ◽  
Conidial Suspension ◽  
Conidial Production ◽  
Plastic Bags ◽  
General Decline ◽  
Leguminous Seeds ◽  
Solid Fermentation

Production of conidia of Penicillium oxalicum (ATCC number pending), a biocontrol agent of Fusarium oxysporum f. sp. lycopersici, was tested in liquid and solid fermentation. P. oxalicum produced 250-fold more conidia in solid than in liquid fermentation at 30 days after inoculation of substrate. Solid fermentation was carried out in plastic bags (600 cm3) especially designed for solid fermentation (VALMIC) containing 50 g of peat/vermiculite (PV) (1:1, wt/wt) with 40% moisture, sealed, sterilized, and then inoculated with 1 ml of a conidial suspension of P. oxalicum (105 conidia g-1 dry substrate), sealed again, and incubated in darkness at 20 to 25°C for 30 days. Addition of amendments to PV in a proportion of 0.5 (wt/wt) significantly increased conidial production of P. oxalicum. The best production was obtained on PV plus meal of cereal grains (barley) or leguminous seeds (lentil) (100-fold higher). Conidial production obtained after 5 days of inoculation was similar to that obtained at 30 days. However, viability of conidia produced in PV plus lentil meal was 35% higher than that of conidia produced in PV plus barley meal. Changes in proportions (1:1:0.5, wt/wt/wt; 1:1:1, wt/wt/wt; 1:0.5:0.5, wt/wt/wt; 1:1:0.5, vol/vol/vol) of components of the substrate (peat/vermiculite/lentil meal) did not enhance production or viability of conidia. Optimal initial moisture in the substrate was 30 to 40%. At lower moistures, significant reductions of production of conidia were observed, particularly at 10%. There was a general decline in the number of conidia in bags with time of storage at -80, -20, 4, and 25°C, or at room temperature (range from 30 to 15°C), with the highest decline occurring from 60 to 180 days. Conidial viability also was reduced with time, except for conidia stored at -20°C. Fresh conidia produced in solid fermentation system or those conidia stored at -20°C for 180 days reduced Fusarium wilt of tomato by 49 and 61%, respectively.

scholarly journals Factors Influencing the Chlorine Susceptibility of Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum

2003 ◽  
Vol 69 (9) ◽  
pp. 5685-5689 ◽  
Author(s):  
Joseph O. Falkinham
Keyword(s):  
Growth Rate ◽  
Mycobacterium Avium ◽  
Growth Phase ◽  
Growth Rates ◽  
Growth Stage ◽  
Tween 80 ◽  
Culture Conditions ◽  
C Cells ◽  
Mycobacterium Intracellulare ◽  
Factors Influencing

ABSTRACT The susceptibility of representative strains of Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum (the MAIS group) to chlorine was studied to identify factors related to culture conditions and growth phase that influenced susceptibility. M. avium and M. intracellulare strains were more resistant to chlorine than were strains of M. scrofulaceum. Transparent and unpigmented colony variants were more resistant to chlorine than were their isogenic opaque and pigmented variants (respectively). Depending on growth stage and growth rate, MAIS strains differed in their chlorine susceptibilities. Cells from strains of all three species growing in early log phase at the highest growth rates were more susceptible than cells in log and stationary phase. Rapidly growing cells were more susceptible to chlorine than slowly growing cells. The chlorine susceptibility of M. avium cells grown at 30°C was increased when cells were exposed to chlorine at 40°C compared to susceptibility after exposure at 30°C. Cells of M. avium grown in 6% oxygen were significantly more chlorine susceptible than cells grown in air. Chlorine-resistant MAIS strains were more hydrophobic and resistant to Tween 80, para-nitrobenzoate, hydroxylamine, and nitrite than were the chlorine-sensitive strains.

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