Production and Partial Purification of Cellulase from a New Isolate, Penicillium verruculosum BS3
Sreedharan Sajith, Sasidharan Sreedevi, Prakasan Priji, Kizhakkepowathial Nair Unni, Sailas Benjamin
Microbiology Research Journal International · pp. 1–12 · Published 6 Jun 2015
10.9734/BMRJ/2015/17865Abstract
Aims: This study describes the production of cellulases by a newly isolated fungus, Penicillium verruculosum BS3 on lignocellulosic biomass with special emphasis on endoglucanase. Study Design: Isolation, screening and characterization of cellulolytic fungus. Production of cellulase by submerged and solid state fermentation. Partial purification of cellulase by ammonium sulphate precipitation, dialysis and spin column. Characterization by Native PAGE and SDS-PAGE. Place and Duration of Study: Enzyme Technology Laboratory, School of Biological Sciences, University of Calicut, Kerala - 673635, India, between October 2013 and November 2014. Methodology: The lignocellulolytic fungus was characterized and identified by lacto-phenol cotton blue staining and 28S rRNA gene sequencing. Preliminary confirmation of cellulase production was done by iodine plate assay method. Production of cellulase was carried out on synthetic and natural raw substrates employing submerged and solid state fermentation strategies. The produced cellulase was purified by ammonium sulphate precipitation, dialysis and spin column followed by characterization using native-PAGE and SDS-PAGE. Results: Primary screening on carboxymethyl cellulose (CMC) agar plates showed cellulase production potential of P. verruculosum BS3. Production of cellulases was quantitatively estimated on basal salt medium (BSM) supplemented with 1% CMC by submerged fermentation (SmF). Among cellulases, endoglucanase showed the highest yield (930.9 U/ml), which was focused on the subsequent studies. The CMC in BSM was replaced by the natural flour (1%) of banana, tapioca, potato or banana peel, of which tapioca flour supported the maximum production of endoglucanase (598.3 U/ml). In solid state fermentation (SSF), P. verruculosum BS3 was grown on sawdust supplemented with 10% banana flour as a solid support, moistened with BSM and the maximum production of endoglucanase (3214.1 U/gds) was observed on day 9 of incubation at 28ºC. The partially purified (9.1 folds purification with 2.1% yield) cellulase was then characterized by native PAGE and SDS-PAGE. Upon staining with Congo-red (1%), on the CMC (0.25%) impregnated native gel, the zymogram showed a major cellulase fraction as yellow opaque zone. From the SDS-PAGE profile, the approximate MW of cellulase was estimated as 17 kDa. Conclusion: This study signifies the industrial importance of the cellulase produced by P. verruculosum BS3 on cheap agro-products, and that the low MW cellulase has added importance as it offers potentials for protein engineering.
References (27)
- 1 PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT [DOI]
- 2 Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 [DOI]
- 3 Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar [DOI]
- 4 Solid state fermentation for the production of industrial enzymes
- 5 A Rapid and Easy Method for the Detection of Microbial Cellulases on Agar Plates Using Gram’s Iodine [DOI]
- 6 Production of cellulases and hemicellulases by Aspergillus niger KK2 from lignocellulosic biomass [DOI]
- 7 Two Cellobiohydrolase-Encoding Genes from Aspergillus niger Require d -Xylose and the Xylanolytic Transcriptional Activator XlnR for Their Expression [DOI]
- 8 Cellulase production by solid state fermentation on lignocellulosic waste from the xylose industry [DOI]
- 9 Production and characterization of cellulolytic enzymes from the thermophilic fungus Thermoascus aurantiacus under solid state cultivation of agricultural wastes [DOI]
- 10 Production of cellulolytic and xylanolytic enzymes by Fusarium oxysporum grown on corn stover in solid state fermentation [DOI]
- 11 Novel cellulases from an extremophilic filamentous fungi Penicillium citrinum: production and characterization [DOI]
- 12 Identification and purification of the main components of cellulases from a mutant strain of Trichoderma viride T 100-14 [DOI]
- 13 Solid-state fermentation of lignocellulosic substrates for cellulase production by Trichoderma reesei NRRL 11460
- 14 SOME PROPERTIES OF CRUDE CARBOXYMTHYL CELLULASE OF ASPERGILLUS NIGERZ10 WILD-TYPE STRAIN
- 15 Improved catalytic efficiency of Endo-β-1,4-glucanase from Bacillus subtilis BME-15 by directed evolution [DOI]
- 16 Cellulases from two Penicillium sp. strains isolated from subtropical forest soil: production and characterization [DOI]
- 17 Differential cellulolytic activity of native-form and C-terminal tagged-form cellulase derived from Coptotermes formosanus and expressed in E. coli [DOI]
- 18 Cellulase production by continuous culture of Trichoderma reesei Rut C30 using acid hydrolysate prepared to retain more oligosaccharides for induction [DOI]
- 19 A novel strain of Trichoderma viride shows complete lignocellulolytic activities [DOI]
- 20 Production and partial purification of cellulase from a novel fungus, Aspergillus flavus BS1 [DOI]
- 21 Cellulase Producing Bacteria from the Wood-Yards on Kallai River Bank [DOI]
- 22 Production of cellulases by Penicillium sp. in a solid-state fermentation of oil palm empty fruit bunch [DOI]
- 23 Lignocellulolytic activities of a novel strain of Trichoderma harzianum [DOI]
- 24 Candida tropicalis BPU1, a novel isolate from the rumen of the Malabari goat, is a dual producer of biosurfactant and polyhydroxybutyrate [DOI]
- 25 Purification and Characterization of Two Low Molecular Weight Endoglucanases Produced by Penicillium occitanis Mutant Pol 6 [DOI]
- 26 Evaluate of pumpkin oil cake as substrate for the cellulase production by Penicillium roqueforti in solid state fermentation
- 27 Illustrated Genera of Imperfect Fungi [DOI]
Cited by 10
Pamella Suely Santa Rosa Pimentel, J. D. De Oliveira, S. Astolfi-Filho · Applied Biochemistry and Biotechnology · 2021
N. Zeb, M. Rashid, B. Aleem · 2019
Hongliang Guo, Xiao-Dong Wang, Duu-Jong Lee · Bioresource Technology · 2018
Ravunni Maya, S. Benjamin · 2016
Hongzhi Bai, Hui Zi, Yuqian Huang · Catalysis Letters · 2017
", ABDULHADI YAKUBU, Department of Microbiology, School of Bioscience and Bioengineering, Lovely Professional University, Phagwara, 144111 Punjab, India" · Cellulose Chemistry and Technology · 2023
Samir Pachpute, Chetan Pawar, Sanjana T. Philip · Techno-Societal 2022 · 2023
Showing 7 of 10 known citations — external sources report more than can currently be individually listed.
Related research
- Biochemical Characteristics of Immobilized Chitinase from Alternaria infectoria — shares topic coverage
- Production, Purification and Characterisation of a Purified Low Molecular Weight and Thermo-alkaline Tolerance Xylanase by Aspergillus brasiliensis — shares topic coverage
- Purification, Characterization and Applications of Proteases Produced by Bacillus amyloliquefaciens 35s Isolated from Soil of the Nile Delta of Egypt — shares topic coverage
- Production, Purification and Characterization of Levan Polymer from Bacillus lentus V8 Strain — shares topic coverage
- Purification, Biochemical Characterization and Applications of Pleurotus ostreatus ARC280 Laccase — shares topic coverage
Article metrics
Real usage data collected on this platform.
80
Page views
0
PDF downloads
60
Outbound clicks
10
Citations
Views over time
Views by country
Approximate, from request IP at view time — not citizenship or institution. Countries with fewer than 5 views are grouped as "Other".
Traffic sources
Referring site, by host.
Views and downloads exclude known bots/crawlers. Citations combines this platform's own DOI-resolved index with each external source's own reported total — see Cited by above for individually listed citing works. Last refreshed 0 seconds ago.