Recombinant production and characterisation of two chitinases from Rasamsonia emersonii, and assessment of their potential industrial applicability

Dwyer, Kelly; Bentley, Ian S; Tighe, Emma; McGleenan, Eibhilin; Gaffney, Darragh; Walsh, Gary

doi:10.1007/s00253-021-11578-3

Cited by 12 publications

(3 citation statements)

References 58 publications

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“…have been correlated with disease suppression [ 63 ], emphasizing the potential of shell-fish waste-based amendments in the manipulation of the soil mycobiome to promote disease suppression. There are several reports on the isolation of chitinases from Talaromyces spp., a highly abundant genus found in the shell degradation consortium [ 64 , 65 , 66 ]. However, as far as we know, the direct involvement of genera Fusicolla , Mycochlamys, and Pseudeurotium in chitin degradation has not been documented.…”

Section: Discussionmentioning

confidence: 99%

Microbial Consortium Associated with Crustacean Shells Composting

2022

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Soil microbes play an essential role in the biodegradation of crustacean shells, which is the process of sustainable bioconversion to chitin derivatives ultimately resulting in the promotion of plant growth properties. While a number of microorganisms with chitinolytic properties have been characterized, little is known about the microbial taxa that participate in this process either by active chitin degradation or by facilitation of this activity through nutritional cooperation and composting with the chitinolytic microorganisms. In this study, we evaluated the transformation of the soil microbiome triggered by close approximation to the green crab shell surface. Our data indicate that the microbial community associated with green crab shell matter undergoes significant specialized changes, which was reflected in a decreased fungal and bacterial Shannon diversity and evenness and in a dramatic alteration in the community composition. The relative abundance of several bacterial and fungal genera including bacteria Flavobacterium, Clostridium, Pseudomonas, and Sanguibacter and fungi Mortierella, Mycochlamys, and Talaromyces were increased with approximation to the shell surface. Association with the shell triggered significant changes in microbial cooperation that incorporate microorganisms that were previously reported to be involved in chitin degradation as well as ones with no reported chitinolytic activity. Our study indicates that the biodegradation of crab shells in soil incorporates a consortium of microorganisms that might provide a more efficient way for bioconversion.

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Section: Discussionmentioning

confidence: 99%

Microbial Consortium Associated with Crustacean Shells Composting

2022

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“…Hence, thermophilic fungi which are capable of secreting high yields of thermostable enzymes have gained considerable interest. One such thermophilic fungus R. emersonii has been reported to contain several lignocelluloytic enzymes including endo-1,4-β-glucanase, β-glucosidase, α-galactosidase, xylanase and amylases [3,5,6], thus possessing great industrial and biotechnological potential.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9 mediated gene editing of transcription factor ACE1 for enhanced cellulase production in thermophilic fungus Rasamsonia emersonii

Singh,

Raheja,

Basotra

et al. 2023

Fungal Biol Biotechnol

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Background The filamentous fungus Rasamsonia emersonii has immense potential to produce biorefinery relevant thermostable cellulase and hemicellulase enzymes using lignocellulosic biomass. Previously in our lab, a hyper-cellulase producing strain of R. emersonii was developed through classical breeding and system biology approaches. ACE1, a pivotal transcription factor in fungi, plays a crucial role in negatively regulating the expression of cellulase genes. In order to identify the role of ACE1 in cellulase production and to further improve the lignocellulolytic enzyme production in R. emersonii, CRISPR/Cas9 mediated disruption of ACE1 gene was employed. Results A gene-edited ∆ACE1 strain (GN11) was created, that showed 21.97, 20.70 and 24.63, 9.42, 18.12%, improved endoglucanase, cellobiohydrolase (CBHI), β-glucosidase, FPase, and xylanase, activities, respectively, as compared to parental strain M36. The transcriptional profiling showed that the expression of global regulator (XlnR) and different CAZymes genes including endoglucanases, cellobiohydrolase, β-xylosidase, xylanase, β-glucosidase and lytic polysaccharide mono-oxygenases (LPMOs) were significantly enhanced, suggesting critical roles of ACE1 in negatively regulating the expression of various key genes associated with cellulase production in R. emersonii. Whereas, the disruption of ACE1 significantly down-regulated the expression of CreA repressor gene as also evidenced by 2-deoxyglucose (2-DG) resistance phenotype exhibited by edited strain GN11 as well as appreciably higher constitutive production of cellulases in the presence of glucose and mixture of glucose and disaccharide (MGDs) both in batch and flask fed batch mode of culturing. Furthermore, ∆ACE1 strains were evaluated for the hydrolysis of biorefinery relevant steam/acid pretreated unwashed rice straw slurry (Praj Industries Ltd; 15% substrate loading rate) and were found to be significantly superior when compared to the benchmark enzymes produced by parent strain M36 and Cellic Ctec3. Conclusions Current work uncovers the crucial role of ACE1 in regulating the expression of the various cellulase genes and carbon catabolite repression mechanism in R. emersonii. This study represents the first successful report of utilizing CRISPR/Cas9 genome editing technology to disrupt the ACE1 gene in the thermophlic fungus R. emersonii. The improved methodologies presented in this work might be applied to other commercially important fungal strains for which genetic manipulation tools are limited.

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“…responsible for the release of N-acetylglucosamine monomers (Fig. 1) (BERINI et al, 2018;DWYER et al, 2021). Furthermore, some fungal β-N-acetylglucosaminidases have already been described as belonging to the GH3 (YANG et al, 2014) and GH20 families (CHEN et al, 2015;QU et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Microbiologia: Geração de conhecimento e caráter multidisciplinar 2

Dias¹

2022

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Editora Direitos para esta edição cedidos à Atena Editora pelos autores. Open access publication by Atena Editora Todo o conteúdo deste livro está licenciado sob uma Licença de Atribuição Creative Commons. Atribuição-Não-Comercial-NãoDerivativos 4.0 Internacional (CC BY-NC-ND 4.0). O conteúdo dos artigos e seus dados em sua forma, correção e confiabilidade são de responsabilidade exclusiva dos autores, inclusive não representam necessariamente a posição oficial da Atena Editora. Permitido o download da obra e o compartilhamento desde que sejam atribuídos créditos aos autores, mas sem a possibilidade de alterála de nenhuma forma ou utilizá-la para fins comerciais.Todos os manuscritos foram previamente submetidos à avaliação cega pelos pares, membros do Conselho Editorial desta Editora, tendo sido aprovados para a publicação com base em critérios de neutralidade e imparcialidade acadêmica.A Atena Editora é comprometida em garantir a integridade editorial em todas as etapas do processo de publicação, evitando plágio, dados ou resultados fraudulentos e impedindo que interesses financeiros comprometam os padrões éticos da publicação. Situações suspeitas de má conduta científica serão investigadas sob o mais alto padrão de rigor acadêmico e ético.

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Recombinant production and characterisation of two chitinases from Rasamsonia emersonii, and assessment of their potential industrial applicability

Cited by 12 publications

References 58 publications

Microbial Consortium Associated with Crustacean Shells Composting

Microbial Consortium Associated with Crustacean Shells Composting

CRISPR/Cas9 mediated gene editing of transcription factor ACE1 for enhanced cellulase production in thermophilic fungus Rasamsonia emersonii

Microbiologia: Geração de conhecimento e caráter multidisciplinar 2

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