scholarly journals SCREENING, ISOLATION, AND PRODUCTION OF FUNGAL LACCASE FROM SAW MILL SOIL OF OSMANABAD.

2020 ◽  
pp. 1-4
Author(s):  
Sabina Shaikh ◽  
Prashant Dixit ◽  
Innus Shaikh

The rainforest and wood decaying habitats are the main sources for the several laccase producers, which include plant, bacteria, fungi and actinomycetes. Among these, several fungal species having laccase producing ability, which mainly includes wood rooting fungi that are often associated with lignin peroxidase or manganese dependent peroxidase or both. In view of this, we screened the natural habitats like saw mill soil and nearby places of Osmanabad district of Maharashtra for isolation of potential fungal species having laccase producing abilities. A total 45 fungal strains were isolated and screened for laccase production on solid media like Czapak dox agar medium containing guaiacol, tannic acid, bromophenol blue, and ABTS. Out of that 24 isolates showed positive results and among these S19 was found to be a potent laccase producing ability. The S19 fungus sp. subcultured on Czapak dox agar slant and screened for laccase enzyme production on solid state fermentation using wheat bran. In conclusion, we isolated the most potent laccase producing S19 fungus sp. from local natural habitats. A total 11.35 U/L of laccase enzyme obtained under optimized solid state fermentation. The enzyme may find potential applications in degradation of xenobiotics, decolourization of dyes, pulp and paper industries, depolymerisation of lignin, pigment degradation, and several other industrial processes.

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3524 ◽  
Author(s):  
Priscila Sutto-Ortiz ◽  
María de los Angeles Camacho-Ruiz ◽  
Manuel R. Kirchmayr ◽  
Rosa María Camacho-Ruiz ◽  
Juan Carlos Mateos-Díaz ◽  
...  

Novel microbial phospholipases A (PLAs) can be found in actinomycetes which have been poorly explored as producers of this activity. To investigate microbial PLA production, efficient methods are necessary such as high-throughput screening (HTS) assays for direct search of PLAs in microbial cultures and cultivation conditions to promote this activity. About 200 strains isolated with selected media for actinomycetes and mostly belonging toStreptomyces(73%) andMicromonospora(10%) genus were first screened on agar-plates containing the fluorophore rhodamine 6G and egg yolk phosphatidylcholine (PC) to detect strains producing phospholipase activity. Then, a colorimetric HTS assay for general PLA activity detection (cHTS-PLA) using enriched PC (≈60%) as substrate and cresol red as indicator was developed and applied; this cHTS-PLA assay was validated with known PLAs. For the first time, actinomycete strains were cultivated by solid-state fermentation (SSF) using PC as inductor and sugar-cane bagasse as support to produce high PLA activity (from 207 to 2,591 mU/g of support). Phospholipase activity of the enzymatic extracts from SSF was determined using the implemented cHTS-PLA assay and the PC hydrolysis products obtained, were analyzed by TLC showing the presence of lyso-PC. Three actinomycete strains of theStreptomycesgenus that stood out for high accumulation of lyso-PC, were selected and analyzed with the specific substrate 1,2-α-eleostearoyl-sn-glycero-3-phosphocholine (EEPC) in order to confirm the presence of PLA activity in their enzymatic extracts. Overall, the results obtained pave the way toward the HTS of PLA activity in crude microbial enzymatic extracts at a larger scale. The cHTS-PLA assay developed here can be also proposed as a routine assay for PLA activity determination during enzyme purification,directed evolution or mutagenesis approaches. In addition, the production of PLA activity by actinomycetes using SSF allow find and produce novel PLAs with potential applications in biotechnology.


2020 ◽  
Vol 42 ◽  
pp. e52699 ◽  
Author(s):  
Alex Graça Contato ◽  
Fabíola Dorneles In´ácio ◽  
Tatiane Brugnari ◽  
Caroline Aparecida Vaz de Araújo ◽  
Giselle Maria Maciel ◽  
...  

Laccases are oxidoreductase enzymes that have the ability to oxidize phenolic substrates. Its biotechnological potential has been greatly explored in many areas as biotechnology industry, bioremediation of dyes, food industry and environmental microbiology. The aim of this study was maximize the laccase production by Pleurotus pulmonarius (Fr.) Quélet in solid-state fermentation (SSF) using orange waste as substrate. After optimization the capability of the crude laccase to decolorize dyes was analyzed. The fermentation medium in the solid-state was optimized by applying a factorial design. After statistics optimization, laccase activity increased two times. The laccase activity appears to be correlated with the ability of crude extract to decolorize some industrial dyes. The optimized laccase was characterized with respect to optimum pH, influence of temperature and salts. Our results demonstrate that P. pulmonarius was an efficient producer of an important industrial enzyme, laccase, in a cheap solid-state system using orange waste as substrate.


2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Susan Grace Karp ◽  
Vincenza Faraco ◽  
Antonella Amore ◽  
Luiz Alberto Junior Letti ◽  
Vanete Thomaz Soccol ◽  
...  

Laccases are oxidative enzymes related to the degradation of phenolic compounds, including lignin units, with concomitant reduction of oxygen to water. Delignification is a necessary pretreatment step in the process of converting plant biomass into fermentable sugars. The objective of this work was to optimize the production of laccases and to evaluate the delignification of sugarcane bagasse byPleurotus ostreatusin solid-state fermentation. Among eight variables (pH, water activity, temperature, and concentrations of CuSO4, (NH4)2SO4, KH2PO4, asparagine, and yeast extract), copper sulfate and ammonium sulfate concentrations were demonstrated to significantly influence laccase production. The replacement of ammonium sulfate by yeast extract and the addition of ferulic acid as inducer provided increases of 5.7- and 2.0-fold, respectively, in laccase activity. Optimization of laccase production as a function of yeast extract, copper sulfate, and ferulic acid concentrations was performed by response surface methodology and optimal concentrations were 6.4 g/L, 172.6 μM, and 1.86 mM, respectively. Experimentally, the maximum laccase activity of 151.6 U/g was produced at the 5th day of solid-state fermentation. Lignin content in sugarcane bagasse was reduced from 31.89% to 26.36% after 5 days and to 20.79% after 15 days by the biological treatment of solid-state fermentation.


2014 ◽  
Vol 1010-1012 ◽  
pp. 42-47
Author(s):  
Jun Yao He ◽  
Xuan Yi Ye ◽  
Qing Zhi Ling ◽  
Li Hui Dong

The production of laccase by solid-state fermentation (SSF) usingArmillariella tabescenswas studied. Wheat bran was selected to be the most suitable solid substrate. Several operational variables including nitrogen source, moisture content, copper and aromatic inducers were investigated. The results showed that the complex nitrogen sources, NH4NO3coupled with peptone was shown to be the best nitrogen source. 75% of initial moisture content was proved to be appropriate. Copper significantly influenced the laccase production and the yield of laccase was improved by addition of 1.5 mM copper sulphate in the medium. Guaiacol efficiently induced the laccase production and the enzyme yield (24500U/g) was enhanced by 32% compared with he control without any aromatic inducers. Efficient production of laccase fromA. tabescenscan be achieved by solid-state fermentation.


Author(s):  
Shradhdha Sharma ◽  
Duggirala Srinivas Murty

Nowadays, a lot of interest has been given to the development of cost-effective and efficient enzyme production technologies. Laccase enzymes are widely used in biotechnological, environmental and industrial sectors. Due to the cost-effectiveness of the solid-state fermentation (SSF) process, it is widely used to produce a broad range of biological products. In this study, optimization of moisture content, temperature, pH, and inoculum size were studied to enhance laccase production ability of Pleurotus sajor-caju in SSF by using One Factor At Time (OFAT) and Response Surface Methodology (RSM). OFAT was used as a baseline study for deducing the experimental design of RSM. The highest production of laccase enzyme (1450 U/g) by Pleurotus sajor-caju on wheat straw was observed at 26°C, 6.0 pH, 72.5 % moisture content, 7.5% inoculum size, 1% fructose and 0.5 % peptone. Unlike the conventional inoculum preparation method, here the inoculum was generated by the spawning method for SSF. The molecular weight of partially purified laccase from Pleurotus sajor-caju was estimated to be around 62 K Da using SDS PAGE. The activity staining of laccase was observed as a zymogram on Native PAGE using ABTS as a substrate. Lignin degradation of wheat straw and its structural disruption due to laccase was observed by Scanning Electron Microscopy (SEM).


Author(s):  
Dalia Sukmawati ◽  
Siti Nurkhasanah ◽  
Zakiah Nur Afifah ◽  
Shabrina Nida Al Husna ◽  
Retno Widowati ◽  
...  

This study reported the application of a next generation sequencing (NGS) analysis of yeast diversity in native Indonesian fruit, Durio Kutejensis, collected from Borneo, Central Kalimantan. The analysis was designed to observe the microbial consortium associated with solid state fermentation (SSF) for amylase production. Together with the additional data from culture-dependent analysis, we observed the morphological features, molecular characteristics, and amylase concentration produced by each isolate. We performed Solid State Fermentation (SSF) for amylase production and the enzyme activity was then determined using UV-Vis spectrophotometer at 540 nm. Result obtained from metagenomic approach consist of 4 group that fungal species included in the Ascomycota identified as Botryosphaeria dothidea (1.35%), Lasiodiplodia crassispora (17.62%), Aureobasidium pullulans (55.02%), Paraphoma chrysanthemicola (11.38%), Preussia funiculate (1.90%), Sporormiella intermedia (0.82%), Myrothecium gramineum (1.35%), Fusarium oxysporum (6.24%), Fusarium proliferatum (3.25%) and Phialemoniopsis curvata (1.08%). The results of isolation using culturable medium in the form of YMA obtained 40 yeast isolates. A total of 40 representative isolates from durian fruit were screened, two positive amylase isolates based on clear zones formed were DU 4.2 (Candida sorboxylosa) and DU4.22 (Cyberlindnera fabianii) isolates with amylolytic index of DU 4.2 isolates at 0.24 and DU 4.22 at 0.72 with an incubation time of 48 h. The highest amylase enzyme activity was found in isolate DU 4.2 of 31.21 U / mL.


BioResources ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. 3017-3031
Author(s):  
Mei-Ling Han ◽  
Qi An ◽  
Kai-Yue Ma ◽  
Wen-Ning An ◽  
Wen-Yao Hao ◽  
...  

The laccase producing abilities of four Basidiomycete fungi species were compared using solid-state fermentation using four different lignocellulosic residues. The biosynthetic potential of the Basidiomycetes was highly dependent on the type of fungi. In general, the laccase secreting ability of Cerrena unicolor Han 849 was greater than Lenzites betulinus Han 851, Stropharia rugosoannulata Han 1321, and Auricularia heimuer Han 1333. The maximum laccase production of C. unicolor Han 849 was approximately 11.25, 122.26, and 15.27 times higher than L. betulinus Han 851, S. rugosoannulata Han 1321 and A. heimuer Han 1333, respectively. Different species of fungi had a preference in lignocellulosic residues. The presence of Firmiana platanifolia was conducive to secreting laccase via C. unicolor Han 849 during solid-state fermentation. A continuous and stable laccase production via C. unicolor Han 849 was an obvious advantage of solid-state fermentation with any of the four lignocellulosic residues used. The maximum laccase production of C. unicolor Han 849 using Firmiana platanifolia was approximately 2.12, 1.68, and 6.13 times higher than Populus beijingensis, Sorghum bicolor, and Oryza sativa, respectively. These findings will be helpful for developing new productivity strains in industrial applications and selecting suitable lignocellulosic residues for laccase production.


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