Browsing by Author "Patrick, Ferdinandi"

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    Optimization of Laccase and Manganese Peroxidase Production in Submerged Culture of Pleurotus Sajor-caju
    (2013) Patrick, Ferdinandi; Mtui, Godliving; Mligo, Anthony M.; Kivaisi, Amelia K.
    A white-rot fungus, Pleurotus sajor-caju, was isolated from coastal Tanzania and screened for crude lignolytic enzymes production using rhemazol brilliant blue R (RBBR) dye, 2,2-azino-bis (3- ethylbenzthiazoline)-6-sulfonate (ABTS) and guaiacol in a semi-solid medium. Laccase (Lac) and manganese peroxidase (MnP) were detected by α-napthol and pyrogallol solutions, respectively, on the guaiacol supplemented semi-solid media. The effect of temperature, pH, carbon, nitrogen, Cu2+, 2,5- xylidine, ferulic acid, Mn2+ and immobilization using Luffa cylindrica sponges in submerged culture fermentations were investigated for maximum enzymes production. After 7 days of incubation, 83 to 100% oxidation of RBBR, ABTS and guaiacol was observed. With optimized culture conditions, the fungal filtrate had maximum Lac and MnP activities of 80 and 0.94 U/ml, respectively compared to 0.62 and 0.0003 U/ml obtained with non-optimized ones; amounting to 129 and 3133 times increase in Lac and MnP activities, respectively. The improved crude enzymes activities, RBBR decolourization, ABTS and guaiacol oxidation capabilities of P. sajor-caju show its potential as a source of industrial enzymes for biotechnological applications.
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    Optimized Production of Lignin Peroxidase, Manganese Peroxidase and Laccase in Submerged Cultures of Trametes Trogii Using Various Growth Media Compositions
    (2010) Patrick, Ferdinandi; Mtui, G.; Mshandete, Anthony M.; Kivaisi, Amelia K.
    A white-rot fungus, Trametes trogii, was isolated from coastal Tanzania and screened for crude lignolytic enzymes production using Rhemazol Brilliant blue R (RBBR) dye, 2,2-azino-bis (3- ethylbenzthiazoline)-6-sulfonate (ABTS) and guaiacol in a semi-solid medium. Lignin peroxidase (LiP), manganese peroxidase (MnP) and Laccase (Lac) were detected by pyrogallol and !- napthol solutions, respectively on the guaiacol supplemented solid media. The effect of temperature, pH, carbon, nitrogen, Cu2+, 2,5-xylidine, ferulic acid, varatryl alcohol and Mn2+ in submerged culture fermentations were investigated for maximum enzymes production. After 7 days of incubation, 72-100% oxidation of RBBR, ABTS and guaiacol was observed. With optimized culture conditions, the fungal filtrate had maximum LiP, MnP and Lac activities of 0.18, 4.44 and 593 U/ml, respectively compared to 0.0011, 0.0054 and 2.3 U/ml obtained with non-optimized ones, amounting to 16,264%, 82,122% and 25,683% increase in LiP, MnP and Lac activities, respectively. The enhanced crude enzymes activities, RBBR decolorization and ABTS guaiacol oxidation capabilities of T. trogii show its potential as a source of industrial enzymes for biotechnological applications.
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    Purification and Characterization of a Laccase from the Basidiomycete Funalia Trogii (Berk.) Isolated in Tanzania
    (2009) Patrick, Ferdinandi; Mtui, Godliving; Mshandete, Anthony M.; Johansson, Gunnar; Kivaisi, Amelia K.
    A lignolytic basidiomycete fungus, Funalia trogii (Berk.), was isolated from decayed wood in coastal Tanzania and cultivated in submerged culture. Initially screened crude enzyme filtrate showed complete rhemazol brilliant blue - R (RBBR) decolorization 2,2’-azino-bis (3-ethylbenzthiazoline)-6-sulfonate and guaiacol oxidation after 7 days of incubation. The fungal filtrate had maximum laccase activity of 593 U/ml after 15 days of incubation. A laccase was purified by anion exchange and size exclusion chromatography to good purity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF). The isolated main component had a molecular weight of ca 58 kDa as determined by MS and an isoelectric point (pI) of 3.8. The optimal pH and temperature range for the purified laccase were 4.0 - 5.0 and 50 - 70 ºC, respectively, using 2, 6- dimethoxyphenol as a substrate.