Article ; Online: Extracellular β-d-fructofuranosidase from a novel Aspergillus sp. DHE1 with high potential for biotechnological applications: Purification and biochemical characterization
Biocatalysis and Agricultural Biotechnology. 2023 Feb. 06, p.102644-
2023 , Page(s) 102644–
Abstract: The current work focused on the purification and biochemical characterization of an extracellular β-d-fructofuranosidase from a novel fungus identified genetically using 18S rRNA molecular sequencing as Aspergillus sp. DHE1 cultured under solid state ... ...
| Abstract | The current work focused on the purification and biochemical characterization of an extracellular β-d-fructofuranosidase from a novel fungus identified genetically using 18S rRNA molecular sequencing as Aspergillus sp. DHE1 cultured under solid state fermentation using the Cotton (Gossypium herbaceum) seed waste as substrate. The enzyme was purified through fractionation by ammonium sulpate salt, ion-exchange chromatography on DEAE-cellulose, and Sephadex G-100 gel filtration, yielding a 13.3-purification fold, 22.6% recovery and 192.9 U/mg protein specific activity. The apparent molecular mass of Aspergillus sp. DHE1 β-d-fructofuranosidase was 78 kDa as estimated on sodium dodecyl sulphate polyacrylamide gel electrophoresis. Optimal pH and temperature were observed at 6.0 and 50 °C, respectively, and the enzyme was quite thermostable at neutral/alkalophilic conditions, retaining 72% of its original activity after 1 h of incubation at 60 °C. The activity of the purified Aspergillus sp. DHE1 β-d-fructofuranosidase was stimulated by 1 mM of KCl (138.5%), NaCl (125.9%), CoCl₂ (121.2%), and CaCl₂ (112.3%), whereas the same concentration of AgCl, FeCl₃, HgCl₂, and CuCl₂ inhibited the activity by 48.7%, 30.5%, 29.9%, and 26.2%, respectively. The kinetic studies using d-sucrose as the substrate were investigated using a Lineweaver-Burk plot, with Kₘ and Vₘₐₓ of 0.85 mM and 47.62 U/mL, respectively, indicating the high substrate affinity of β-d-fructofuranosidase. The ability of Aspergillus sp. DHE1 β-d-fructofuranosidase to tolerate ethanol suggests that it may have numerous uses in the food industry, including the production of alcoholic beverages. |
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| Keywords | Aspergillus ; Gossypium herbaceum ; agricultural biotechnology ; ammonium ; biocatalysis ; cotton ; enzymes ; ethanol ; food industry ; fractionation ; fungi ; gel chromatography ; ion exchange chromatography ; molecular weight ; pH ; polyacrylamide gel electrophoresis ; sodium dodecyl sulfate ; solid state fermentation ; temperature ; thermal stability ; wastes ; Aspergillus sp. DHE1 ; β-d-fructofuranosidase ; Cotton seed waste ; Purification ; Biochemical characterization ; Ethanol tolerance |
| Language | English |
| Dates of publication | 2023-0206 |
| Publishing place | Elsevier Ltd |
| Document type | Article ; Online |
| Note | Pre-press version |
| ZDB-ID | 2642052-1 |
| ISSN | 1878-8181 |
| ISSN | 1878-8181 |
| DOI | 10.1016/j.bcab.2023.102644 |
| Database | NAL-Catalogue (AGRICOLA) |
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