Partial Purification and Characterization of Endoxylanase from a fungus, Leohumicola incrustata
- Authors: Adeoyo, Olusegun R , Pletschke, Brett I , Dames, Joanna F
- Date: 2021
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/440393 , vital:73779 , 10.4314/br.v19i1.2
- Description: Xylanases are glycoside hydrolases (GH) that degrade β-1, 4-xylan, a linear polysaccharide found as hemicellulose in cell wall of plants. Endoxylanase (Endo-1, 4-β-xylanase, EC 3.2. 1.8) randomly catalyses xylan to produce varying short xylooligosaccharides (XOS). This study aimed to determine the characteristics of a partially purified endoxylanase from Leohumicola incrustata. Enzyme production was carried out using beechwood (BW) xylan, after which the cell-free crude filtrate was concentrated using the ammonium sulphate precipitation method. The hydrolysed products were analysed by thin-layer chromatography (TLC) and zymography. The result showed that the enzyme produced varying smaller-sized linear xylooligosaccharides with R f values corresponding to those of xylobiose, xylotriose, xylotetraose, xylopentaose, xylohexaose and other higher oligomers. The endoxylanase had a molecular mass of 72 kDa. The enzyme is stable in the presence of K+, Na+, Ca 2+, Fe 2+, Mg 2+, Zn 2+, Co 2+, pH of 5.0 and temperature of 37 o C. However, the activity gradually decreased after 60 min at 50 o C and retained over 69% activity after 120 min, while at 60 and 70 o C, the enzyme activity sharply decreased (pre-incubation periods). Endoxylanase from L. incrustata is comparable to those of other microorganisms and should be considered an attractive candidate for future industrial applications.
- Full Text:
- Date Issued: 2021
Molecular identification and antibacterial properties of an ericoid associated mycorrhizal fungus
- Authors: Adeoyo, Olusegun R , Pletschke, Brett I , Dames, Joanna F
- Date: 2019
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/440378 , vital:73778 , https://doi.org/10.1186/s12866-019-1555-y
- Description: The quest for novel sources of antibacterial compounds have necessitated the inclusion of ericoid mycorrhizal fungi (ERM) commonly found within the root of ericaceous plants. Agar-well diffusion method was used to detect antibacterial activity and was followed by the microbroth diffusion method [minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)].
- Full Text:
- Date Issued: 2019
Purification and characterization of an amyloglucosidase from an ericoid mycorrhizal fungus (Leohumicola incrustata)
- Authors: Adeoyo, Olusegun R , Pletschke, Brett I , Dames, Joanna F
- Date: 2018
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/440405 , vital:73780 , https://doi.org/10.1186/s13568-018-0685-1
- Description: This study aimed to purify and characterize amyloglucosidase (AMG) from Leohumicola incrustata. AMG was purified to homogeneity from cell-free culture filtrate of an ERM fungus grown in a modified Melin–Norkrans liquid medium. The molecular mass of the AMG was estimated to be 101 kDa by combining the results of Sephadex G-100 gel filtration, sodium dodecyl sulphate–polyacrylamide gel electrophoresis, and zymography. The Km and kcat values were 0.38 mg mL−1 and 70 s−1, respectively, using soluble starch as a substrate. The enzyme was stable at 45 °C (pH 5.0), retaining over 65% activity after a pre-incubation period of 24 h. The metal inhibition profile of the AMG showed that Mn2+ and Ca2+ enhanced activity, while it was stable to metals ions, except a few (Al3+, Co2+, Hg2+ and Cd2+) that were inhibitory at a concentration higher than 5 mM. Thin layer chromatography revealed that only glucose was produced as the product of starch hydrolysis. The amylase from L. incrustata is a glucoamylase with promising characteristics such as temperature stability over an extended period, high substrate affinity and stability to a range of chemicals. Also, this study reports for the first time the possibility of using some culturable ERM fungi to produce enzymes for the bio-economy.
- Full Text:
- Date Issued: 2018
Improved endoglucanase production and mycelial biomass of some ericoid fungi
- Authors: Adeoyo, Olusegun R , Pletschke, Brett I , Dames, Joanna F
- Date: 2017
- Language: English
- Type: article , text
- Identifier: http://hdl.handle.net/10962/61435 , vital:28026 , https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5209310/
- Description: Fungal species associated with ericaceous plant roots produce a number of enzymes and other bio-active metabolites in order to enhance survival of their host plants in natural environments. This study focussed on endoglucanase production from root associated ericoid mycorrhizal and dark septate endophytic fungal isolates. Out of the five fungal isolates screened, Leohumicola sp. (ChemRU330/PPRI 13195) had the highest relative enzyme activity and was tested along with isolates belonging to Hyloscyphaceae (EdRU083/PPRI 17284) and Leotiomycetes (EdRU002/PPRI 17261) for endoglucanase production under different pH and nutritional conditions that included: carbon sources, nitrogen sources and metal ions, at an optimum temperature of 28 °C. An optimal of pH 5.0 produced enzyme activity of 3.99, 2.18 and 4.31 (U/mg protein) for isolates EdRU083, EdRU002 and Leohumicola sp. respectively. Increased enzyme activities and improved mycelial biomass production were obtained in the presence of supplements such as potassium, sodium, glucose, maltose, cellobiose, tryptone and peptone. While NaFe-EDTA and Co2+ inhibited enzyme activity. The potential role of these fungi as a source of novel enzymes is an ongoing objective of this study.
- Full Text:
- Date Issued: 2017
Improved endoglucanase production and mycelial biomass of some ericoid fungi
- Authors: Adeoyo, Olusegun R , Pletschke, Brett I , Dames, Joanna F
- Date: 2017
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/440352 , vital:73776 , https://doi.org/10.1186/s13568-016-0312-y
- Description: Fungal species associated with ericaceous plant roots produce a number of enzymes and other bio-active metabolites in order to enhance survival of their host plants in natural environments. This study focussed on endoglucanase production from root associated ericoid mycorrhizal and dark septate endophytic fungal isolates. Out of the five fungal isolates screened, Leohumicola sp. (ChemRU330/PPRI 13195) had the highest relative enzyme activity and was tested along with isolates belonging to Hyloscyphaceae (EdRU083/PPRI 17284) and Leotiomycetes (EdRU002/PPRI 17261) for endoglucanase production under different pH and nutritional conditions that included: carbon sources, nitrogen sources and metal ions, at an optimum temperature of 28 °C. An optimal of pH 5.0 produced enzyme activity of 3.99, 2.18 and 4.31 (U/mg protein) for isolates EdRU083, EdRU002 and Leohumicola sp. respectively. Increased enzyme activities and improved mycelial biomass production were obtained in the presence of supplements such as potassium, sodium, glucose, maltose, cellobiose, tryptone and peptone. While NaFe-EDTA and Co2+ inhibited enzyme activity. The potential role of these fungi as a source of novel enzymes is an ongoing objective of this study.
- Full Text:
- Date Issued: 2017