Agrobacterium-mediated delivery of marker genes to Phanerochaete chrysosporium mycelial pellets: a model transformation system for white-rot fungi

2006 ◽  
Vol 43 (3) ◽  
pp. 181 ◽  
Author(s):  
Shefali Gupta ◽  
Krishna Kant Sharma ◽  
Ramesh Chander Kuhad

Genetics provides an approach to the analysis of the complex function of lignin biodegradation, through the isolation of mutants and the creation of gene libraries for the identification of genes and their products. However, white-rot fungi (for example, Phanerochaete chrysosporium ) have not so far been analysed from this point of view, and there is the challenge of establishing such genetics. P. chrysosporium is convenient experimentally because relatively few genes are switched on at the onset of ligninolytic activity. We describe the isolation of clones carrying genes expressed specifically in the ligninolytic phase, the development of a general strategy for mapping such clones, and the elucidation of the mating system of this organism. Another objective is the development of methods for transforming DNA into P. chrysosporium . This would allow the use of site-directed mutagenesis to analyse the functioning of ligninases, and the control of expression of the corresponding genes. The use of genetic crosses for strain improvement and the identification of components of the system are also discussed.


2020 ◽  
Vol 21 (2) ◽  
pp. 416
Author(s):  
Angel De La Cruz Pech-Canul ◽  
Javier Carrillo-Campos ◽  
María de Lourdes Ballinas-Casarrubias ◽  
Rosa Lidia Solis-Oviedo ◽  
Selena Karina Hernández-Rascón ◽  
...  

Manganese peroxidases (MnP) from the white-rot fungi Phanerochaete chrysosporium catalyse the oxidation of Mn2+ to Mn3+, a strong oxidizer able to oxidize a wide variety of organic compounds. Different approaches have been used to unravel the enzymatic properties and potential applications of MnP. However, these efforts have been hampered by the limited production of native MnP by fungi. Heterologous expression of MnP has been achieved in both eukaryotic and prokaryotic expression systems, although with limited production and many disadvantages in the process. Here we described a novel molecular approach for the expression and purification of manganese peroxidase isoform 1 (MnP1) from P. chrysosporium using an E. coli-expression system. The proposed strategy involved the codon optimization and chemical synthesis of the MnP1 gene for optimised expression in the E. coli T7 shuffle host. Recombinant MnP1 (rMnP1) was expressed as a fusion protein, which was recovered from solubilised inclusion bodies. rMnP1 was purified from the fusion protein using intein-based protein purification techniques and a one-step affinity chromatography. The designated strategy allowed production of an active enzyme able to oxidize guaiacol or Mn2+.


2016 ◽  
Vol 75 (1) ◽  
Author(s):  
Laksmita Prima SANTI ◽  
Lisdar Idwan SUDIRMAN ◽  
Didiek Hadjar GOENADI

SummaryFungal treatment by using white-rot fungito reduce a wide variety of herbicide com-pounds is a specialized bioremediation pro-cess. A laboratory experiment was conductedto determine the ability of Phanerochaetechrysosporium, Ceriporiopsis subvermispora,and Pleurocybella porrigens and seven white-rot fungi isolated from a native of tropicalenvironment to grow on yeast malt extractglucose (YMG) agar containing highconcentration of (I) 2,4-dichlorophenoxy aceticacid, (R) glyphosate, and (G) paraquat. Thedata indicated that P. chrysosporium couldgrow on YMG media containing 5000 ppm of(I) 2,4-D, whereas BPBPI 02/04 isolate onYMG 250 ppm of (R) glyphosate or (G)paraquat. Relative values of growth inhibitionof these fungi are 81.1; 27.8; and 50.0%respectively. Biodegradation capability ofherbicides by candidate inoculants in soil-sandmedia was also determined in greenhouseexperiment by using peanut, sorghum, corn,and Borreria alata as bio-indicators. Peanutand B. alata were found to be the bestresponsive seedlings as bio-indicator on thepresence of (I) 2,4-D herbicide in soil-sandmedia.RingkasanTeknologi bioremediasi dengan fungipelapuk putih (FPP) digunakan untuk me-reduksi sejumlah senyawa herbisida. Kegiatanpenelitian yang dilakukan di laboratoriumbertujuan untuk mengetahui kemampuan tum-buh Phanerochaete chrysosporium, Ceripo-riopsis subvermispora, dan Pleurocybellaporrigens serta tujuh isolat FPP yang diperolehdari lingkungan tropik secara in vitro padamedium agar yeast malt extract glucose(YMG) yang mengandung (I) 2,4-dikloro-fenoksi asam asetat, (R) glifosat, dan (G)parakuat konsentrasi tinggi. Dari data yangdiperoleh, diketahui bahwa Ph. chrysosporiummemiliki kemampuan tumbuh dalam mediumpadat YMG yang mengandung 5000 ppm (I)2,4-D dan isolat BPBPI 02/04 pada 250 ppm(R) glifosat dan (G) parakuat dengan nilaihambatan pertumbuhan relatif terhadap kontrol(HPR) masing-masing 81,1; 27,8; dan 50,0%.Pengujian isolat terpilih terhadap kemampuanmendegradasi herbisida di dalam mediumtanah dan pasir juga dilakukan di rumah kacadengan menggunakan kacang tanah, sorgum,jagung, dan Boreria alata sebagai bioindikator.Kacang tanah dan B. alata memberikan responterbaik terhadap keberadaan herbisida (I) 2,4-Ddi dalam medium tanah dan pasir .


PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0247452
Author(s):  
Micol Bellucci ◽  
Francesca Marazzi ◽  
Alida Musatti ◽  
Riccardo Fornaroli ◽  
Andrea Turolla ◽  
...  

The treatability of seven wastewater samples generated by a textile digital printing industry was evaluated by employing 1) anammox-based processes for nitrogen removal 2) microalgae (Chlorella vulgaris) for nutrient uptake and biomass production 3) white-rot fungi (Pleurotus ostreatus and Phanerochaete chrysosporium) for decolorization and laccase activity. The biodegradative potential of each type of organism was determined in batch tests and correlated with the main characteristics of the textile wastewaters through statistical analyses. The maximum specific anammox activity ranged between 0.1 and 0.2 g N g VSS-1 d-1 depending on the sample of wastewater; the photosynthetic efficiency of the microalgae decreased up to 50% during the first 24 hours of contact with the textile wastewaters, but it improved from then on; Pleurotus ostreatus synthetized laccases and removed between 20–62% of the colour after 14 days, while the enzymatic activity of Phanerochaete chrysosporium was inhibited. Overall, the findings suggest that all microbes have great potential for the treatment and valorisation of textile wastewater after tailored adaptation phases. Yet, the depurative efficiency can be probably enhanced by combining the different processes in sequence.


2021 ◽  
Vol 11 (21) ◽  
pp. 9930
Author(s):  
Ana Isabel Díaz ◽  
Marta Ibañez ◽  
Adriana Laca ◽  
Mario Díaz

The liquid fraction from the two-phase extraction process in the olive industry (alperujo), is a waste that contains lignocellulosic organic matter and phenolic compounds, difficult to treat by conventional biological methods. Lignocellulosic enzymes from white-rot fungi can be an interesting solution to break down these recalcitrant compounds and advance the treatment of that waste. In the present work the ability of Phanerochaete chrysosporium to degrade the abovementioned liquid waste (AL) was studied. Experiments were carried out at 26 °C within the optimal pH range 4–6 for 10 days and with and without the addition of glucose, measuring the evolution of COD, BOD5, biodegradability index, reducing sugars, total phenolic compounds, and colour. The results obtained in this study revealed the interest of Phanerochaete chrysosporium for an economical and eco-friendly treatment of alperujo, achieving COD and colour removals around 60%, and 32% of total phenolic compounds degradation, regardless of glucose addition.


1983 ◽  
Vol 29 (4) ◽  
pp. 457-463 ◽  
Author(s):  
A. E. Reade ◽  
R. E. McQueen

Five isolates of wood-rotting fungi were compared for their ability to increase the digestibility of poplar shavings. Homogenates of the fungi were inoculated into mixtures of 25 g of shavings and 100 mL of supplementary nutrient solution and incubated at 25 °C. The in vitro rumen digestibility of the products increased from 30% at the start of the fermentation to maxima of 72% with Polyporus anceps after 8 weeks, 64% with Ganoderma applanatum and 62% with Phanerochaete chrysosporium after 4 weeks, 61% with Polyporus versicolor after 3 weeks, and 42% with Fomitopsis ulmarius after 4 weeks. Fibre analysis showed a decrease in lignin as digestibility increased. Loss of carbohydrates occurred in all fermentations and continued after lignin loss ceased. Dry matter yield at the time of maximum digestibility ranged from 80 to 94% of the original dry matter. The highest digestibilities attained were similar to those of conventional roughage feeds for ruminants. This shows that this fermentation could form the basis of a practical system for converting poplar into a feedstuff.


Sign in / Sign up

Export Citation Format

Share Document