Aims and Scope
Recent Articles
Functional and Structural Characterization of a Novel Isoamylase from Ostreococcus tauri and Role of the N-Terminal Domain
Nicolas Hedín, Julieta Barchiesi, Diego F. Gomez-Casati, María V. Busi
Background:
The debranching starch enzymes, isoamylase 1 and 2 are well-conserved enzymes present in almost all the photosynthetic organisms. These enzymes are involved in the crystallization process of starch and are key components which remove misplaced α-1,6 ramifications on the final molecule.
Aim:
In this work, we performed a functional and structural study of a novel isoamylase from Ostreococcus tauri.
Methods:
We identified conserved amino acid residues possibly involved in catalysis. We also identified a region at the N-terminal end that resembles a Carbohydrate Binding Domain (CBM), which is more related to the family CBM48, but has no spatial conservation of the residues involved in carbohydrate binding.
Results:
The cloning, expression and biochemical characterization of this N-terminal region confirmed that it binds to polysaccharides, showing greater capacity for binding to amylopectin rather than total starch or amylose.
Conclusion:
This module could be a variant of the CBM48 family or it could be classified within a new CBM family.
February 12, 2020
Articles
- February 13, 2020
Effect of Plant Spacing and Np Fertilizer Levels on Growth, Seed Yield and Quality of Onion (Allium cepa L.) at Shewa Robit, Northern Ethiopia
February 13, 2020
Upregulation of Antioxidant Gene Expressions and Enzyme Activity Against Acrylamide-Induced Neurotoxicity in Mice after Grape Seed Extract Treatment
March 29, 2020Citric Acid Production by the Solid-State Cultivation Consortium of Aspergillus Niger and Trichoderma Reesei from Sugarcane Bagasse
Editor's Choice
Use of Aqueous Two-Phase and Three-Phase Partitioning Systems for Purification of Lipase Obtained in Solid-State Fermentation by Rhizopus arrhizus
Valentina Dobreva, Boriana Zhekova, Georgi Dobrev
Background:
Purification of enzymes by conventional methods such as precipitation and chromatographic techniques is a costly and time-consuming procedure and may lead to low yields of enzyme activity. Alternative liquid-liquid extraction methods such as Aqueous Two-Phase Systems (ATPS) and Three Phase Partitioning (TPP) are characterized by the high enzyme yields and purification degree.
Objective:
The objective of this study was the application of partitioning systems ATPS and TPP for purification of lipase produced in solid-state fermentation by Rhizopus arrhizus.
Methods:
ATPS and TPP were used for purification of lipase, obtained by solid state cultivation of Rhizopus arrhizus.
Results:
Lipase was isolated with PEG4000/potassium sodium tartrate ATPS and the effect of the system composition, including PEG 4000 and potassium sodium tartrate concentrations on lipase partitioning was studied. When using 30% PEG4000/21% potassium sodium tartrate, lipase was distributed in the top phase, and the highest recovery yield of 217% and purification fold of 6.1 were achieved. It was found that at PEG4000 concentration of or higher than 15%, the enzyme was present in the top polymer-rich phase with a partitioning yield of over 90%. Upon application of TPP for lipase isolation, the effect of t-butanol concentration, ammonium sulfate concentration and pH on enzyme partitioning was investigated. The highest lipase recovery yield of 71% and 19.1-fold purification were achieved in the interfacial phase in the presence of 30% ammonium sulfate saturation with 1.0:0.5 crude extract/t-butanol ratio at pH 7 in a single step. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymographic analysis showed significant purification of lipase by TPP and the presence of two multiple forms of the enzyme.
Conclusion:
ATPS (PEG4000/ Potassium sodium tartrate) and TPP (1.0:0.5 crude extract/t-butanol ratio, 30% ammonium sulfate saturation, pH 7) proved to be rapid methods for the isolation and purification of lipase and they can be used in downstream processing for industrial preparation of the enzyme.
April 24, 2019
Other Post
- August 30, 2018
Immobilization of Cholesterol Oxidase: An Overview
April 28, 2017
Evaluation of Fixed-Bed Cultures with Immobilized Lactococcus Lactis ssp. Lactis on Different Scales
August 11, 2017
Effects of the Addition of Sodium Chloride to a Tetrameric Protein in Water Solution During Exposure to High Frequency Electromagnetic Field
September 21, 2017
Molecular Identification of Yeasts and Lactic Acid Bacteria Involved in the Production of Beninese Fermented Food Degue
