A trimeric and thermostable lichenase from B. pumilus US570 strain: Biochemical and molecular characterization

Elgharbi, Fatma; Hlima, Hajer Ben; Ameri, Rihab; Béjar, Samír; Hmida-Sayari, Aïda

doi:10.1016/j.ijbiomac.2016.11.021

Cited by 12 publications

(9 citation statements)

References 29 publications

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“…β-1,3–1,4-glucanase purification was also achieved by other several chromatographic procedures including affinity chromatography (Apiraksakorn et al 2008 ; Kim et al 2013 ), HPLC using a gel filtration column (Elgharbi et al 2013 ) and Ni-IDA column (Zhang et al 2017 ). Although the non- Bacillus β-1,3–1,4-glucanases are usually larger than the Bacillus enzymes because of additional domains (Planas 2000 ), Halomonas meridiana ES021 β-1,3–1,4-glucanase enzyme has a molecular mass about 72 kDa which was similar to that purified from Bacillus pumilus US570 (75 kDa) (Elgharbi et al 2017 ). It was smaller than other non-Bacillus β-1,3–1,4-glucanases as reported by Ruminococcus flavofaciens (Flint et al 1993 ) and Pseudoalteromonas (Nakatani et al 2012 ) with molecular masses about 90 kDa.…”

Section: Discussionmentioning

confidence: 95%

Purification, characterization, immobilization and applications of an enzybiotic β-1,3–1,4-glucanase produced from halotolerant marine Halomonas meridiana ES021

Gadallah

El-Borai

El-Aassar

et al. 2023

World J Microbiol Biotechnol

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Extracellular β-1,3–1,4-glucanase-producing strain Halomonas meridiana ES021 was isolated from Gabal El-Zeit off shore, Red Sea, Egypt. The Extracellular enzyme was partially purified by precipitation with 75% acetone followed by anion exchange chromatography on DEAE-cellulose, where a single protein band was determined with molecular mass of approximately 72 kDa. The Km value was 0.62 mg β-1,3–1,4-glucan/mL and Vmax value was 7936 U/mg protein. The maximum activity for the purified enzyme was observed at 40 °C, pH 5.0, and after 10 min of the reaction. β-1,3–1,4-glucanase showed strong antibacterial effect against Bacillus subtilis, Streptococcus agalactiae and Vibrio damsela. It also showed antifungal effect against Penicillium sp. followed by Aspergillus niger. No toxicity was observed when tested on Artemia salina. Semi-purified β-1,3–1,4-glucanase was noticed to be effective in clarification of different juices at different pH values and different time intervals. The maximum clarification yields were 51.61% and 66.67% on mango juice at 40 °C and pH 5.3 for 2 and 4 h, respectively. To our knowledge, this is the first report of β-1,3–1,4-glucanase enzyme from halotolerant Halomonas species. Graphic Abstract

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Section: Discussionmentioning

confidence: 95%

Purification, characterization, immobilization and applications of an enzybiotic β-1,3–1,4-glucanase produced from halotolerant marine Halomonas meridiana ES021

Gadallah

El-Borai

El-Aassar

et al. 2023

World J Microbiol Biotechnol

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“…Of note, the CelA203 activity was maintained at over 80% after the purified CelA203 was stored at 4°C for over one year. We reported stronger thermostability of CelA203 than BGlc8H [ 38 ], Egl-257 [ 39 ], and lichenase EG1 [ 37 ], but lower than that compared to lichenase UEB-S [ 10 ] and the glucanase from Bacillus pumilus US570 [ 40 ]. Furthermore, 1 mM Zn 2+ , Cu 2+ , Fe 3+ , Mn 2+ , and 10 mM EDTA significantly inhibited the activity of CelA203, agreeing with the results reported of β-1,3-1,4-glucanase from Bacillus altitudinis YC-9 [ 12 ].…”

Section: Discussionmentioning

confidence: 99%

“…Elsewhere, Cu 2+ , Mn 2+ , and EDTA were found to reduce the activity of β-1,3-1,4-glucanase from B. subtilis MA139 [ 41 ]. The specific activity of CelA203 towards barley β-glucan (566.38 U/mg) and lichenan (171.61 U/mg) was higher than that of GluUS570 (161.87 U/mg and 70.38 U/mg) [ 8 ] , and PbBglu16A (425 U/mg and 212 U/mg) [ 14 ], but lower than that of TaGlu34 (13,527 U/mg and 9,225 U/mg) [ 17 ]. For barley β-glucan, the K m value of CelA203(3.98 mg/ml) was similar with that of Egl7A (4 mg/ml) [ 13 ] and AaBglu12A (3.51 mg/ml) [ 18 ], but the V max value (1,017.17 U/mg) was lower than that of both (17,951 U/mg and 12,068 U/mg respectively).…”

Section: Discussionmentioning

confidence: 99%

“…Microbial lichenases belong to glycoside hydrolase family 16 [ 7 ]. Previous investigations have isolated, cloned, and characterized a number of lichenases from Bacillus species, including B. pumilus [ 8 ], B. subtilis [ 9 - 11 ], B. altitudinis [ 12 ], and B. tequilensis [ 13 ]. In addition, several non- Bacillus species have been studied, including Paenibacillus barengoltzii [ 14 ], Paenibacillus barcinonensis BP-23 [ 15 ], Penicillium occitanis Pol6 [ 16 ], Thermoascus aurantiacus [ 17 ], Aspergillus awamori [ 18 ], and Aspergillus niger US368 [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

Huang

Wang

et al. 2022

J. Microbiol. Biotechnol.

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Lichenase is an enzyme mainly implicated in the degradation of polysaccharides in the cell walls of grains. Emerging evidence shows that a highly efficient expression of a thermostable recombinant lichenase holds considerable promise for application in the beer-brewing and animal feed industries. Herein, we cloned a lichenase gene (CelA203) from Bacillus subtilis B110 and expressed it in E. coli. This gene contains an ORF of 729 bp, encoding a protein with 242 amino acids and a calculated molecular mass of 27.3 kDa. According to the zymogram results, purified CelA203 existed in two forms, a monomer, and a tetramer, but only the tetramer had potent enzymatic activity. CelA203 remained stable over a broad pH and temperature range and retained 40% activity at 70°C for 1 h. The K m and V max of CelA203 towards barley β-glucan and lichenan were 3.98 mg/ml, 1017.17 U/mg, and 2.78 mg/ml, 198.24 U/mg, respectively. Furthermore, trisaccharide and tetrasaccharide were the main products obtained from CelA203-mediated hydrolysis of deactivated oat bran. These findings demonstrate a promising role for CelA203 in the production of oligosaccharides in animal feed and brewing industries.

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“…Recently, it was observed that mutations located not only inside but also outside the protein interfaces introduce conformational changes that can alter the oligomeric balance (the transition from monomers to oligomers) by creating new bonds, or through the indirect stabilization of protein dynamics, again suggesting a relation between higher-order oligomeric states and thermostability [9,10]. Furthermore, although mesophilic proteins show less thermostability than their thermophilic homologs, it has been observed that protein-protein interactions allow both the homo-oligomeric [11,12] and hetero-oligomeric [13] organization of a wide variety of mesophilic enzymes, which play an important role in many biological pathways and even in cell-cell adhesion processes. As an example, it was recently demonstrated that the reduction in the oligomeric state, through the substitution of conserved amino acid residues, served to abolish the high thermostability of enzymes from pathogenic microorganisms [14].…”

Section: Introductionmentioning

confidence: 99%

Contribution of the Oligomeric State to the Thermostability of Isoenzyme 3 from Candida rugosa

et al. 2018

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Thermophilic proteins have evolved different strategies to maintain structure and function at high temperatures; they have large, hydrophobic cores, and feature increased electrostatic interactions, with disulfide bonds, salt-bridging, and surface charges. Oligomerization is also recognized as a mechanism for protein stabilization to confer a thermophilic adaptation. Mesophilic proteins are less thermostable than their thermophilic homologs, but oligomerization plays an important role in biological processes on a wide variety of mesophilic enzymes, including thermostabilization. The mesophilic yeast Candida rugosa contains a complex family of highly related lipase isoenzymes. Lip3 has been purified and characterized in two oligomeric states, monomer (mLip3) and dimer (dLip3), and crystallized in a dimeric conformation, providing a perfect model for studying the effects of homodimerization on mesophilic enzymes. We studied kinetics and stability at different pHs and temperatures, using the response surface methodology to compare both forms. At the kinetic level, homodimerization expanded Lip3 specificity (serving as a better catalyst on soluble substrates). Indeed, dimerization increased its thermostability by more than 15 °C (maximum temperature for dLip3 was out of the experimental range; >50 °C), and increased the pH stability by nearly one pH unit, demonstrating that oligomerization is a viable strategy for the stabilization of mesophilic enzymes.

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A trimeric and thermostable lichenase from B. pumilus US570 strain: Biochemical and molecular characterization

Cited by 12 publications

References 29 publications

Purification, characterization, immobilization and applications of an enzybiotic β-1,3–1,4-glucanase produced from halotolerant marine Halomonas meridiana ES021

Purification, characterization, immobilization and applications of an enzybiotic β-1,3–1,4-glucanase produced from halotolerant marine Halomonas meridiana ES021

Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

Contribution of the Oligomeric State to the Thermostability of Isoenzyme 3 from Candida rugosa

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