A novel ionic liquid-stable halophilic endoglucanase from Stachybotrys microspora

Hmad, Ines Ben; Boudabbous, Manel; Belghith, Hafedh; Gargouri, Ali

doi:10.1016/j.procbio.2017.01.007

Cited by 31 publications

(15 citation statements)

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“…Although ILs are also known to denature enzymes, there are some reports of endoglucanases which are stable towards ILs. The endoglucanases from Stachybotrys microspora are 50% active in the presence of 20% (v/v) 1-butyl-3-methylimidazolium chloride [ 59 ]. Gladden et al reported an endoglucanase from GH12 with the highest activity in the presence of 15% (v/v)…”

Section: Discussionmentioning

confidence: 99%

Genome-wide characterization of cellulases from the hemi-biotrophic plant pathogen, Bipolaris sorokiniana, reveals the presence of a highly stable GH7 endoglucanase

Aich

Singh

Kundu

et al. 2017

Biotechnol Biofuels

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Background Bipolaris sorokiniana is a filamentous fungus that causes spot blotch disease in cereals like wheat and has severe economic consequences. However, information on the identities and role of the cell wall-degrading enzymes (CWDE) in B. sorokiniana is very limited. Several fungi produce CWDE like glycosyl hydrolases (GHs) that help in host cell invasion. To understand the role of these CWDE in B. sorokiniana, the first step is to identify and annotate all possible genes of the GH families like GH3, GH6, GH7, GH45 and AA9 and then characterize them biochemically.ResultsWe confirmed and annotated the homologs of GH3, GH6, GH7, GH45 and AA9 enzymes in the B. sorokiniana genome using the sequence and domain features of these families. Quantitative real-time PCR analyses of these homologs revealed that the transcripts of the BsGH7-3 (3rd homolog of the GH 7 family in B. sorokiniana) were most abundant. BsGH7-3, the gene of BsGH7-3, was thus cloned into pPICZαC Pichia pastoris vector and expressed in X33 P. pastoris host to be characterized. BsGH7-3 enzyme showed a temperature optimum of 60 °C and a pHopt of 8.1. BsGH7-3 was identified to be an endoglucanase based on its broad substrate specificity and structural comparisons with other such endoglucanases. BsGH7-3 has a very long half-life and retains 100% activity even in the presence of 4 M NaCl, 4 M KCl and 20% (v/v) ionic liquids. The enzyme activity is stimulated up to fivefold in the presence of Mn+2 and Fe+2 without any deleterious effects on enzyme thermostability.ConclusionsHere we reanalysed the B. sorokiniana genome and selected one GH7 enzyme for further characterization. The present work demonstrates that BsGH7-3 is an endoglucanase with a long half-life and no loss in activity in the presence of denaturants like salt and ionic liquids, and lays the foundation towards exploring the Bipolaris genome for other cell wall-degrading enzymes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0822-0) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Genome-wide characterization of cellulases from the hemi-biotrophic plant pathogen, Bipolaris sorokiniana, reveals the presence of a highly stable GH7 endoglucanase

Aich

Singh

Kundu

et al. 2017

Biotechnol Biofuels

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“…Biochemical studies on S. microspora have shown a halophilic character of secreted endoglucanases [ 19 , 20 ]. Moreover, some strains form Stachybotrys genera are considered a marine fungus like S. chartarum according to the World Register of Marine Species ( https://fr.wikipedia.org/wiki/Stachybotrys#WRMS ), S. longispora [ 33 ], and Stachybotrys sp.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to its richness in β‐glucosidases, our fungus is distinguished by the very specific properties of its endoglucanases. Indeed, we have shown that it secretes two enzymes, called EG1 and EG2, which are very interesting for their high salt tolerance [ 19 , 20 ]. Indeed, EG1 is the most halophilic fungal endoglucanase ever studied, with an optimal activity at 5.6 m NaCl and resists SDS by retaining more than 70% of its activity at 10% SDS [ 19 ].…”

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confidence: 99%

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Large‐scale analysis of the genome of the rare alkaline‐halophilic Stachybotrys microspora reveals 46 cellulase genes

et al. 2023

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Fungi are of great importance in biotechnology, for example in the production of enzymes and metabolites. The main goal of this study was to obtain a high‐coverage draft of the Stachybotrys microspora genome and to annotate and analyze the genome sequence data. The rare fungus S. microspora N1 strain is distinguished by its ability to grow in an alkaline halophilic environment and to efficiently secrete cellulolytic enzymes. Here we report the draft genome sequence composed of 3715 contigs, a genome size of 35 343 854 bp, with a GC content of 53.31% and a coverage around 20.5×. The identification of cellulolytic genes and of their corresponding functions was carried out through analysis and annotation of the whole genome sequence. Forty‐six cellulases were identified using the fungicompanion bioinformatic tool. Interestingly, an S. microspora endoglucanase selected from those with a low isoelectric point was predicted to have a halophilic profile and share significant homology with a well‐known bacterial halophilic cellulase. These results confirm previous biochemical studies revealing a halophilic character, which is a very rare feature among fungal cellulases. All these properties suggest that cellulases of S. microspora may have potential for use in the biofuel, textile, and detergent industries.

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“…4). On the contrary, the activity of a halophilic endoglucanase that was enhanced by NaCl was not stimulated by ILs (Ben Hmad et al 2017 ). This suggests a different mechanism of enzyme activation by ILs and that other types of IL-protein interactions are important, in addition to electrostatic effects.…”

Section: Resultsmentioning

confidence: 99%

Enhanced activity of hyperthermostable Pyrococcus horikoshii endoglucanase in superbase ionic liquids

Hebal

Hämäläinen²,

Makkonen

et al. 2022

Biotechnol Lett

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Objectives Ionic liquids (ILs) that dissolve biomass are harmful to the enzymes that degrade lignocellulose. Enzyme hyperthermostability promotes a tolerance to ILs. Therefore, the limits of hyperthemophilic Pyrococcus horikoschii endoglucanase (PhEG) to tolerate 11 superbase ILs were explored. Results PhEG was found to be most tolerant to 1-ethyl-3-methylimidazolium acetate ([EMIM]OAc) in soluble 1% carboxymethylcellulose (CMC) and insoluble 1% Avicel substrates. At 35% concentration, this IL caused an increase in enzyme activity (up to 1.5-fold) with CMC. Several ILs were more enzyme inhibiting with insoluble Avicel than with soluble CMC. Km increased greatly in the presence ILs, indicating significant competitive inhibition. Increased hydrophobicity of the IL cation or anion was associated with the strongest enzyme inhibition and activation. Surprisingly, PhEG activity was increased 2.0–2.5-fold by several ILs in 4% substrate. Cations exerted the main role in competitive inhibition of the enzyme as revealed by their greater binding energy to the active site. Conclusions These results reveal new ways to design a beneficial combination of ILs and enzymes for the hydrolysis of lignocellulose, and the strong potential of PhEG in industrial, high substrate concentrations in aqueous IL solutions.

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A novel ionic liquid-stable halophilic endoglucanase from Stachybotrys microspora

Cited by 31 publications

References 42 publications

Genome-wide characterization of cellulases from the hemi-biotrophic plant pathogen, Bipolaris sorokiniana, reveals the presence of a highly stable GH7 endoglucanase

Genome-wide characterization of cellulases from the hemi-biotrophic plant pathogen, Bipolaris sorokiniana, reveals the presence of a highly stable GH7 endoglucanase

Large‐scale analysis of the genome of the rare alkaline‐halophilic Stachybotrys microspora reveals 46 cellulase genes

Enhanced activity of hyperthermostable Pyrococcus horikoshii endoglucanase in superbase ionic liquids

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