Record ID No. |
5572 |
Author(s) |
Adeoyo O. R.*, Pletschke B. I. and Dames J. F. , 2018 |
Affiliation |
*Department of Biochemistry and Microbiology, Rhodes University, P.O. Box 94, Grahamstown, 6140, South Africa. |
Title |
Purification and characterization of an amyloglucosidase from an ericoid mycorrhizal fungus (Leohumicola incrustata). |
Source. Vol.(no):Page |
AMB Express. 8(1): 154, doi: 10.1186/s13568-018-0685-1. |
Categories |
|
Subjects |
Methodology |
Sub-subjects |
Carbohydrate metabolism |
Host |
NA |
Organism |
Leohumicola incrustata |
Country |
South Africa, Nigeria. |
Abstracts |
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. |