Recent advancements in biorefinery processes necessitate search for cost effective and thermostable cellulases. This study was designed to characterize the cellulase obtained from a thermophilic bacterium, Neobacillus sedimentimangrovi UE25. A combined pretreatment of NaOH and methyltrioctylammonium chloride was given to sugarcane bagasse (SB) for lignin removal and the pretreated SB was utilized as a carbon source for the cellulase production. The thermostable cellulase thus obtained was characterized by adopting central composite design which has not been reported earlier for this purpose. Cellulase showed its maximum activity at pH 7 and temperature 60 ℃ and it remained active in the presence of many salts and detergents. Endoglucanase (EG) was found to be stable for 30 min at 80 ℃. The purification of EG by using DEAE column yielded specific activity and purification fold of 365.866 IU mg and 4.264, respectively. The purified EG had a molecular weight of ∼45 kDa. End product tolerance of EG was also evident, as an activity of 228.57 IU mL was observed in the presence of 60 mM glucose which revealed that it does not lose its activity upon accumulation of end-product when the reaction is prolonged. The purified EG exhibited V and K of 294 U mL min and 36 µM, respectively, in the presence of 60 mM glucose. This novel thermostable cellulase can finds its applications in industrial sector.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.enzmictec.2022.110133 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
New-Generation Antibacterial Agent-Cellulose-Binding Thermostable TP84_Endolysin.
Int J Mol Sci
December 2024
Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, 80-309 Gdansk, Poland.
The increasing antibiotic resistance among bacteria challenges the biotech industry to search for new antibacterial molecules. Endolysin TP84_28 is a thermostable, lytic enzyme, encoded by the bacteriophage (phage) TP-84, and it effectively digests host bacteria cell wall. Biofilms, together with antibiotic resistance, are major problems in clinical medicine and industry.
View Article and Find Full Text PDFSimultaneous enhancement of activity and stability of Bacillus safensis-derived laccase and its application in lignocellulose saccharification.
Bioresour Technol
December 2024
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR China. Electronic address:
Effective hydrolysis of lignocelluloses for producing reducing sugar is impeded by the covalent binding of hemicellulose and cellulose through lignin, which could be eliminated by laccases. This study identified a novel thermostable laccase from Bacillus safensis TCCC 111022 and created an iterative mutant E231D/Y441H, exhibiting 1.59-fold greater specific activity and a 183 % greater half-life at 80°C than the wild-type enzyme.
View Article and Find Full Text PDFThermobifida fusca Cel6B moves bidirectionally while processively degrading cellulose.
Biotechnol Biofuels Bioprod
December 2024
Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, 37235, USA.
Background: Cellulose, an abundant biopolymer, has great potential to be utilized as a renewable fuel feedstock through its enzymatic degradation into soluble sugars followed by sugar fermentation into liquid biofuels. However, crystalline cellulose is highly resistant to hydrolysis, thus industrial-scale production of cellulosic biofuels has been cost-prohibitive to date. Mechanistic studies of enzymes that break down cellulose, called cellulases, are necessary to improve and adapt such biocatalysts for implementation in biofuel production processes.
View Article and Find Full Text PDFArtificial trimerization of β-glucosidase for enhanced thermostability and activity via computational redesign.
Int J Biol Macromol
January 2025
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China. Electronic address:
The improvement of enzyme thermostability is of great significance in the biotechnology industry. Herein, we modified the C-terminus of β-glucosidase via the computer-aided rational design and successfully obtained six variants that exhibit improved optimal temperatures and thermal inactivation half-lives at 65 °C. Among these variants, the thermal inactivation half-life and hydrolytic activity of TrCel1b-H13 at 65 °C were increased by 416 and 3223 folds, respectively, compared to those of the wild type.
View Article and Find Full Text PDFTwo-step computational redesign of Bacillus subtilis cellulase and β-glucanase for enhanced thermostability and activity.
Int J Biol Macromol
January 2025
AIM center, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address:
The growing demand for biocatalysts in biomass processing highlights the necessity of enhancing the thermostability of glycoside hydrolases. However, improving both thermostability and activity is often hindered by trade-offs between backbone rigidity and the flexibility of substrate-binding regions. In this study, Bacillus subtilis cellulase and β-glucanase were engineered using a two-step process incorporating the computational tools Pythia and ESM-2, which were found complementary in improving stability and activity.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!