Abstract
BackgroundCellulose is the most prevalent biomass and renewable energy source in nature. The hydrolysis of cellulosic biomass to glucose units is essential for the economic exploitation of this natural resource. Cellulase enzyme, which is largely generated by bacteria and fungus, is commonly used to degrade cellulose. Cellulases are used in a variety of industries, including bioethanol manufacturing, textiles, detergents, drugs, food, and paper. As part of our quest to find an efficient biocatalyst for the hydrolysis of cellulosic biomass, we describe the amplification, cloning, and sequencing of cellulase (cel9z) from Bacillus licheniformis strain Z9, as well as the characterization of the resulting enzyme. ResultsCellulase was partially purified from B. licheniformis strain Z9 using (NH4)2SO4 precipitation and Sephadex G-100 gel column chromatography with 356.5 U/mg specific activity, 2.1-purification fold, and 3.07 % yield. The nucleotide sequence of the cellulase gene was deposited to the GenBank, B. licheniformis strain Z9 cellulase (cel9z) gene, under accession number MK814929. This corresponds to 1453 nucleotides gene and encodes for a protein composed of 484 amino acids. Comparison of deduced amino acids sequence to other related cellulases showed that the enzyme cel9z can be classified as a glycoside hydrolase family 9. SDS-PAGE analysis of the purified enzyme revealed that the molecular mass was 54.5 kDa. The optimal enzyme activity was observed at pH 7.4 and 30 °C. The enzyme was found to be strongly inhibited by Mg2+ and Na+, whereas strongly activated by Fe3+, Cu2+, and Ca2+. ConclusionsB. licheniformis strain Z9 and its cellulase gene can be further utilized for recombinant production of cellulases for industrial application.
Highlights
Cellulose is the most prevalent biomass and renewable energy source in nature
Molecular identification of cellulase gene A total volume of 50 μL of Polymerase chain reaction (PCR) reaction was used for amplification of cellulase cel9z from B. licheniformis strain Z9
The PCR products of strain Z9 were sequenced to obtain an open reading frame (ORF) of 1453-bp gene sequence, which was consistent with the result of electrophoresis
Summary
Cellulose is the most prevalent biomass and renewable energy source in nature. The hydrolysis of cel‐ lulosic biomass to glucose units is essential for the economic exploitation of this natural resource. Cellulase enzyme, which is largely generated by bacteria and fungus, is commonly used to degrade cellulose. As part of our quest to find an efficient biocatalyst for the hydrolysis of cellulosic biomass, we describe the amplification, cloning, and sequencing of cellulase (cel9z) from Bacillus licheniformis strain Z9, as well as the characterization of the resulting enzyme. Cellulose is the most prevalent polysaccharide in nature and the primary component of plant cell walls [1]. Due to the urgent need for green energy, cellulose has acquired economic interest in its hydrolysis bio-technique in recent decades. One of the most common methods for converting cellulose into reducing sugars, which can be turned into ethanol and other compounds, is microbial hydrolysis [3]
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