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Dörschmann, P., Kopplin, G., Thalenhorst, T., Seeba, C., Ullah, S. F., Srivastava, V., . . . Klettner, A. (2025). Influence of a Very High-Molecular Weight Fucoidan from Laminaria hyperborea on Age-Related Macular Degeneration-Relevant Pathomechanisms in Ocular Cell Models. Marine Drugs, 23(3), Article ID 101.
Open this publication in new window or tab >>Influence of a Very High-Molecular Weight Fucoidan from Laminaria hyperborea on Age-Related Macular Degeneration-Relevant Pathomechanisms in Ocular Cell Models
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2025 (English)In: Marine Drugs, E-ISSN 1660-3397, Vol. 23, no 3, article id 101Article in journal (Refereed) Published
Abstract [en]

Fucoidans from Laminaria hyperborea (LH) can be antioxidative, antiangiogenic, and anti-inflammatory. In this study, a very high-molecular weight (3700 kDa) fucoidan from LH, FucBB04, was tested regarding its bioactivity in age-related macular degeneration (AMD) models in vitro. Primary retinal pigment epithelium (RPE) from pig eyes, human uveal melanoma cell line OMM-1, and RPE cell line ARPE-19 were used. Substituents of the extract were determined with chemical analysis. Cell viability was tested with tetrazolium assay (MTT), oxidative stress was induced by H2O2 or erastin, respectively. Secreted vascular endothelial growth factor A (VEGF-A) was assessed with ELISA. Retinal pigment epithelium 65 kDa protein (RPE65) and protectin (CD59) protein expression were tested in Western blot. Cell barrier was assessed by measuring trans-epithelial electrical resistance (TEER), phagocytic ability by a fluorescence assay. Gene expression and secretion of interleukin 6 (IL-6) and interleukin 8 (IL-8) were tested in real-time PCR and ELISA. FucBB04 displayed no oxidative stress protective effects. Its effect on VEGF was inconsistent, with VEGF secretion reduced in primary RPE, but not in ARPE-19. On the other hand, Lipopolysaccharide (LPS) and polyinosinic/polycytidylic acid (PIC)-induced IL-6 or IL-8 secretion was reduced by FucBB04, while complement inhibiting protein CD59 was not affected. In addition, FucBB04 did not influence the gene expression of IL-6 or IL-8. Visual cycle protein RPE65 expression, phagocytic ability, and barrier function were reduced by FucBB04. Very high-molecular weight fucoidan from LH shows bioactivities against AMD-related pathological pathways, but adverse effects on RPE function may limit its suitability as a therapeutic compound. Smaller high-molecular weight fucoidans are recommended for further research.

Place, publisher, year, edition, pages
MDPI AG, 2025
Keywords
fucoidan, gene expression, interleukin, phagocytosis, polyinosinic/polycytidylic acid (PIC), protectin (CD59), retinal pigment epithelium-specific 65 kDa protein (RPE65), sulfated fucan, toll-like receptor, trans-epithelial electrical resistance (TEER), vascular endothelial growth factor (VEGF)
National Category
Ophthalmology Neurosciences
Identifiers
urn:nbn:se:kth:diva-362231 (URN)10.3390/md23030101 (DOI)001452483400001 ()40137287 (PubMedID)2-s2.0-105001122574 (Scopus ID)
Note

QC 20250425

Available from: 2025-04-09 Created: 2025-04-09 Last updated: 2025-04-25Bibliographically approved
Ullah, S. F., Oreb, M., Boles, E., Srivastava, V., Seidl-Seiboth, V., Seiboth, B. & Kappel, L. (2025). N-acetylglucosamine sensing in the filamentous soil fungus Trichoderma reesei. The FEBS Journal
Open this publication in new window or tab >>N-acetylglucosamine sensing in the filamentous soil fungus Trichoderma reesei
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2025 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658Article in journal (Refereed) Epub ahead of print
Abstract [en]

N-acetylglucosamine (GlcNAc) is involved in diverse signaling pathways in dimorphic yeasts and bacteria and is related to morphogenetic switching, mating, stress, virulence, and cell death. Recently, GlcNAc has been shown to promote plant growth by shaping the bacterial soil community. However, the role of GlcNAc sensing in filamentous soil fungi has not been investigated. By using Trichoderma reesei as a model organism, we show here that GlcNAc impacts the expression of around 2100 genes. Carbohydrate metabolism, amino acid metabolism, and secondary metabolism were the three most strongly affected classes of eukaryotic orthologous groups (KOG classes). Two key regulators of GlcNAc catabolism, the NDT80 domain-containing transcriptional regulator RON1, and a GlcNAc sensor, NGS1, are needed for differential regulation of two-thirds of these genes. In silico structural modeling of NGS1 identified a domain with homology to the GCN5-related histone acetyltransferase from Candida albicans, which serves as a GlcNAc catabolism regulator and GlcNAc sensor. Finally, we characterized the third regulator of GlcNAc sensing in T. reesei, which is the highly specific GlcNAc transporter N-acetylglucosamine transporter (NGT1). Using a deletion mutant of ngt1, we demonstrate that GlcNAc has to enter the cell to activate the GlcNAc catabolic gene expression. Interestingly, in contrast to dimorphic yeasts, the pathways for defense and pathogenicity seem to be induced in T. reesei by external GlcNAc. Given the ancestral role of Trichoderma spp. in the fungal kingdom and the highly conserved GlcNAc catabolism cluster that includes their regulators in many species of fungi, we propose a regulatory network for GlcNAc sensing in soil fungi.

Place, publisher, year, edition, pages
Wiley, 2025
Keywords
major facilitator superfamily, <italic>N</italic>-acetylglucosamine, secondary metabolism, signaling, soil fungi, <italic>Trichoderma</italic> spp.
National Category
Microbiology
Identifiers
urn:nbn:se:kth:diva-360762 (URN)10.1111/febs.70015 (DOI)001421422600001 ()39954246 (PubMedID)2-s2.0-85219699737 (Scopus ID)
Note

QC 20250303

Available from: 2025-03-03 Created: 2025-03-03 Last updated: 2025-05-27Bibliographically approved
Lee, Y. J., Kim, W. R., Park, E. G., Lee, D. H., Kim, J.-m., Jeong, H.-s., . . . Kim, H.-S. (2025). Phenotypic and Gene Expression Alterations in Aquatic Organisms Exposed to Microplastics. International Journal of Molecular Sciences, 26(3), Article ID 1080.
Open this publication in new window or tab >>Phenotypic and Gene Expression Alterations in Aquatic Organisms Exposed to Microplastics
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2025 (English)In: International Journal of Molecular Sciences, ISSN 1661-6596, E-ISSN 1422-0067, Vol. 26, no 3, article id 1080Article, review/survey (Refereed) Published
Abstract [en]

The use of plastics, valued for its affordability, durability, and convenience, has grown significantly with the advancement of industry. Paradoxically, these very properties of plastics have also led to significant environmental challenges. Plastics are highly resistant to decomposition, resulting in their accumulation on land, where they eventually enter aquatic environments, due to natural processes or human activities. Among these plastics, microplastics, which are tiny plastic particles, are particularly concerning when they enter aquatic ecosystems, including rivers and seas. Their small size makes them easily ingestible by aquatic organisms, either by mistake or through natural feeding behaviors, which poses serious risks. Moreover, microplastics readily adsorb other pollutants present in aquatic environments, creating pollutant complexes that can have a synergistic impact, magnifying their harmful effects compared to microplastics or pollutants acting alone. As a result, extensive research has focused on understanding the effects of microplastics on aquatic organisms. Numerous studies have demonstrated that aquatic organisms exposed to microplastics, either alone or in combination with other pollutants, exhibit abnormal hatching, development, and growth. Additionally, many genes, particularly those associated with the antioxidant system, display abnormal expression patterns in these conditions. In this review, we examine these impacts, by discussing specific studies that explore changes in phenotype and gene expression in aquatic organisms exposed to microplastics, both independently and in combination with adsorbed pollutants.

Place, publisher, year, edition, pages
MDPI AG, 2025
Keywords
aquatic organism, gene expression, heavy metal, microplastic, organic compound, phenotype, pollutant, qPCR, sequencing, water
National Category
Environmental Sciences
Identifiers
urn:nbn:se:kth:diva-360819 (URN)10.3390/ijms26031080 (DOI)001418689200001 ()39940847 (PubMedID)2-s2.0-85217642271 (Scopus ID)
Note

QC 20250303

Available from: 2025-03-03 Created: 2025-03-03 Last updated: 2025-03-03Bibliographically approved
Fassolo, E. M., Rosa, S., Marciano, D., Tagliani, A., Feni, L., Srivastava, V., . . . Pesaresi, P. (2024). AMP32: a short cyclic peptide as an environmentally friendly alternative to counteract late blight symptoms in crops. Journal of Peptide Science, 30
Open this publication in new window or tab >>AMP32: a short cyclic peptide as an environmentally friendly alternative to counteract late blight symptoms in crops
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2024 (English)In: Journal of Peptide Science, ISSN 1075-2617, E-ISSN 1099-1387, Vol. 30Article in journal, Meeting abstract (Other academic) Published
Place, publisher, year, edition, pages
WILEY, 2024
National Category
Other Natural Sciences
Identifiers
urn:nbn:se:kth:diva-355804 (URN)001314146400208 ()
Note

QC 20241104

Available from: 2024-11-04 Created: 2024-11-04 Last updated: 2024-11-04Bibliographically approved
Kao, M.-R., Parker, J., Oehme, D., Chang, S.-C., Cheng, L.-C., Wang, D., . . . Hsieh, Y. S. Y. (2024). Substrate Specificities of Variants of Barley (1,3)- and (1,3;1,4)-β-d-Glucanases Resulting from Mutagenesis and Segment Hybridization. Biochemistry, 63(9), 1194-1205
Open this publication in new window or tab >>Substrate Specificities of Variants of Barley (1,3)- and (1,3;1,4)-β-d-Glucanases Resulting from Mutagenesis and Segment Hybridization
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2024 (English)In: Biochemistry, ISSN 0006-2960, E-ISSN 1520-4995, Vol. 63, no 9, p. 1194-1205Article in journal (Refereed) Published
Abstract [en]

Barley (1,3;1,4)-β-d-glucanase is believed to have evolved from an ancestral monocotyledon (1,3)-β-d-glucanase, enabling the hydrolysis of (1,3;1,4)-β-d-glucans in the cell walls of leaves and germinating grains. In the present study, we investigated the substrate specificities of variants of the barley enzymes (1,3;1,4)-β-d-glucan endohydrolase [(1,3;1,4)-β-d-glucanase] isoenzyme EII (HvEII) and (1,3)-β-d-glucan endohydrolase [(1,3)-β-d-glucanase] isoenzyme GII (HvGII) obtained by protein segment hybridization and site-directed mutagenesis. Using protein segment hybridization, we obtained three variants of HvEII in which the substrate specificity was that of a (1,3)-β-d-glucanase and one variant that hydrolyzed both (1,3)-β-d-glucans and (1,3;1,4)-β-d-glucans; the wild-type enzyme hydrolyzed only (1,3;1,4)-β-d-glucans. Using substitutions of specific amino acid residues, we obtained one variant of HvEII that hydrolyzed both substrates. However, neither protein segment hybridization nor substitutions of specific amino acid residues gave variants of HvGII that could hydrolyze (1,3;1,4)-β-d-glucans; the wild-type enzyme hydrolyzed only (1,3)-β-d-glucans. Other HvEII and HvGII variants showed changes in specific activity and their ability to degrade the (1,3;1,4)-β-d-glucans or (1,3)-β-d-glucans to larger oligosaccharides. We also used molecular dynamics simulations to identify amino-acid residues or structural regions of wild-type HvEII and HvGII that interact with (1,3;1,4)-β-d-glucans and (1,3)-β-d-glucans, respectively, and may be responsible for the substrate specificities of the two enzymes.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2024
National Category
Biochemistry Molecular Biology
Identifiers
urn:nbn:se:kth:diva-346237 (URN)10.1021/acs.biochem.3c00673 (DOI)001200213500001 ()38598309 (PubMedID)2-s2.0-85190095966 (Scopus ID)
Funder
Australian Research Council, CE110001007The Swedish Foundation for International Cooperation in Research and Higher Education (STINT), KO2018-7936
Note

QC 20240508

Available from: 2024-05-07 Created: 2024-05-07 Last updated: 2025-02-20Bibliographically approved
de Oliveira, A. S., de Oliveira, J. S., Kumar, R., Silva, F. B., Fernandes, M. R., Nobre, F. D., . . . Brilhante, R. S. (2023). Antifungal activity of sustainable histone deacetylase inhibitors against planktonic cells and biofilms of Candida spp. and Cryptococcusneoformans. Medical Mycology, 61(8), Article ID myad073.
Open this publication in new window or tab >>Antifungal activity of sustainable histone deacetylase inhibitors against planktonic cells and biofilms of Candida spp. and Cryptococcusneoformans
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2023 (English)In: Medical Mycology, ISSN 1369-3786, E-ISSN 1460-2709, Vol. 61, no 8, article id myad073Article in journal (Refereed) Published
Abstract [en]

The limited therapeutic options for fungal infections and the increased incidence of fungal strains resistant to antifungal drugs, especially Candida spp., require the development of new antifungal drugs and strategies. Histone deacetylase inhibitors (HDACi), like vorinostat, have been studied in cancer treatment and have antifungal effects, acting alone or synergistically with classical antifungals. Here we investigated the antifungal activity of two novel sustainable HDACi (LDT compounds) based on vorinostat structure. Molecular docking simulation studies reveal that LDT compounds can bind to Class-I HDACs of Candida albicans, C. tropicalis, and Cryptococcus neoformans, which showed similar binding mode to vorinostat. LDT compounds showed moderate activity when tested alone against fungi but act synergistically with antifungal azoles against Candida spp. They reduced biofilm formation by more than 50% in C. albicans (4 µg/mL), with the main action in fungal filamentation. Cytotoxicity of the LDT compounds against RAW264.7 cells was evaluated and LDT536 demonstrated cytotoxicity only at the concentration of 200 µmol/L, while LDT537 showed IC50 values of 29.12 µmol/L. Our data indicated that these sustainable and inexpensive HDACi have potential antifungal and antibiofilm activities, with better results than vorinostat, although further studies are necessary to better understand the mechanism against fungal cells.

Place, publisher, year, edition, pages
Oxford University Press (OUP), 2023
Keywords
antifungal activity, HDACi, hydroxamic acid, biofilm
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:kth:diva-334763 (URN)10.1093/mmy/myad073 (DOI)001046486100001 ()37553154 (PubMedID)2-s2.0-85168735561 (Scopus ID)
Note

QC 20230824

Available from: 2023-08-24 Created: 2023-08-24 Last updated: 2024-08-28Bibliographically approved
Srivastava, A., Kumar, A., Biswas, S., Srivastava, V., Rajaram, H. & Mishra, Y. (2023). Cd-induced cytosolic proteome changes in the cyanobacterium Anabaena sp. PCC7120 are mediated by LexA as one of the regulatory proteins. Biochimica et Biophysica Acta - Proteins and Proteomics, 1871(3), 140902, Article ID 140902.
Open this publication in new window or tab >>Cd-induced cytosolic proteome changes in the cyanobacterium Anabaena sp. PCC7120 are mediated by LexA as one of the regulatory proteins
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2023 (English)In: Biochimica et Biophysica Acta - Proteins and Proteomics, ISSN 1570-9639, E-ISSN 1878-1454, Vol. 1871, no 3, p. 140902-, article id 140902Article in journal (Refereed) Published
Abstract [en]

LexA, a well-characterized transcriptional repressor of SOS genes in heterotrophic bacteria, has been shown to regulate diverse genes in cyanobacteria. An earlier study showed that LexA overexpression in a cyanobacterium, Anabaena sp. PCC7120 reduces its tolerance to Cd stress. This was later shown to be due to modulation of photosynthetic redox poising by LexA under Cd stress. However, due to the global regulatory nature of LexA and the prior prediction of AnLexA-box in a few heavy metal-responsive genes, we speculated that LexA has a broad role in Cd tolerance, with regulation over a variety of Cd stress-responsive genes in addition to photosynthetic genes. Thus, to further expand the knowledge on the regulatory role of LexA in Cd stress tolerance, a cytosolic proteome profiling of Anabaena constitutively overexpressing LexA upon Cd stress was performed. The proteomic study revealed 25 differentially accumulated proteins (DAPs) in response to the combined effect of LexA over -expression and Cd stress, and the other 11 DAPs exclusively in response to either LexA overexpression or Cd stress. The 36 identified proteins were related with a variety of functions, including photosynthesis, C -meta-bolism, antioxidants, protein turnover, post-transcriptional modifications, and a few unknown and hypothetical proteins. The regulation of LexA on corresponding genes, and six previously reported Cd efflux transporters, was further validated by the presence of AnLexA-boxes, transcript, and/or promoter analyses. In a nutshell, this study identifies the regulation of Anabaena LexA on several Cd stress-responsive genes of various functions, hence expanding the regulatory role of LexA under Cd stress.

Place, publisher, year, edition, pages
Elsevier BV, 2023
Keywords
Electrophoretic mobility shift assay, LexA, Mass spectrometry, Real-time quantitative PCR, Transcription regulation, Two-dimensional gel electrophoresis
National Category
Other Chemistry Topics
Identifiers
urn:nbn:se:kth:diva-329876 (URN)10.1016/j.bbapap.2023.140902 (DOI)001002048100001 ()36716944 (PubMedID)2-s2.0-85147377815 (Scopus ID)
Note

QC 20230626

Available from: 2023-06-26 Created: 2023-06-26 Last updated: 2023-06-26Bibliographically approved
Rocafort, M., Srivastava, V., Bowen, J. K., Diaz-Moreno, S. M., Guo, Y., Bulone, V., . . . Mesarich, C. H. (2023). Cell Wall Carbohydrate Dynamics during the Differentiation of Infection Structures by the Apple Scab Fungus, Venturia inaequalis. Microbiology Spectrum, 11(3)
Open this publication in new window or tab >>Cell Wall Carbohydrate Dynamics during the Differentiation of Infection Structures by the Apple Scab Fungus, Venturia inaequalis
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2023 (English)In: Microbiology Spectrum, E-ISSN 2165-0497, Vol. 11, no 3Article in journal (Refereed) Published
Abstract [en]

Scab, caused by the biotrophic fungal pathogen Venturia inaequalis, is the most economically important disease of apples. During infection, V. inaequalis colonizes the subcuticular host environment, where it develops specialized infection structures called runner hyphae and stromata. These structures are thought to be involved in nutrient acquisition and effector (virulence factor) delivery, but also give rise to conidia that further the infection cycle. Despite their importance, very little is known about how these structures are differentiated. Likewise, nothing is known about how these structures are protected from host defenses or recognition by the host immune system. To better understand these processes, we first performed a glycosidic linkage analysis of sporulating tubular hyphae from V. inaequalis developed in culture. This analysis revealed that the V. inaequalis cell wall is mostly composed of glucans (44%) and mannans (37%), whereas chitin represents a much smaller proportion (4%). Next, we used transcriptomics and confocal laser scanning microscopy to provide insights into the cell wall carbohydrate composition of runner hyphae and stromata. These analyses revealed that, during subcuticular host colonization, genes of V. inaequalis putatively associated with the biosynthesis of immunogenic carbohydrates, such as chitin and b-1,6-glucan, are downregulated relative to growth in culture, while on the surface of runner hyphae and stromata, chitin is deacetylated to the less-immunogenic carbohydrate chitosan. These changes are anticipated to enable the subcuticular differentiation of runner hyphae and stromata by V. inaequalis, as well as to protect these structures from host defenses and recognition by the host immune system. IMPORTANCE Plant-pathogenic fungi are a major threat to food security. Among these are subcuticular pathogens, which often cause latent asymptomatic infections, making them difficult to control. A key feature of these pathogens is their ability to differentiate specialized subcuticular infection structures that, to date, remain largely understudied. This is typified by Venturia inaequalis, which causes scab, the most economically important disease of apples. In this study, we show that, during subcuticular host colonization, V. inaequalis downregulates genes associated with the biosynthesis of two immunogenic cell wall carbohydrates, chitin and b-1,6-glucan, and coats its subcuticular infection structures with a less-immunogenic carbohydrate, chitosan. These changes are anticipated to enable host colonization by V. inaequalis and provide a foundation for understanding subcuticular host colonization by other plant-pathogenic fungi. Such an understanding is important, as it may inform the development of novel control strategies against subcuticular plant-pathogenic fungi.

Place, publisher, year, edition, pages
American Society for Microbiology, 2023
Keywords
apple scab, cell wall, KEYWORDS Venturia inaequalis, morphological differentiation, subcuticular infection structures
National Category
Other Chemistry Topics
Identifiers
urn:nbn:se:kth:diva-334628 (URN)10.1128/spectrum.04219-22 (DOI)000969407400001 ()37039647 (PubMedID)2-s2.0-85163913746 (Scopus ID)
Note

QC 20230823

Available from: 2023-08-23 Created: 2023-08-23 Last updated: 2023-10-06Bibliographically approved
Chen, X.-R., Wang, Y., Kale, S. D., Fang, Y. & Srivastava, V. (2023). Editorial: Apoplastic effectors - What roles do they play in plant-pathogen interactions?. Frontiers in Microbiology, 14, Article ID 1149771.
Open this publication in new window or tab >>Editorial: Apoplastic effectors - What roles do they play in plant-pathogen interactions?
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2023 (English)In: Frontiers in Microbiology, E-ISSN 1664-302X, Vol. 14, article id 1149771Article in journal, Editorial material (Other academic) Published
Place, publisher, year, edition, pages
Frontiers Media SA, 2023
Keywords
apoplastic effectors, plant pathogens, virulence function, PTI, interaction, evolution
National Category
Biochemistry Molecular Biology
Identifiers
urn:nbn:se:kth:diva-325177 (URN)10.3389/fmicb.2023.1149771 (DOI)000941175500001 ()36865775 (PubMedID)2-s2.0-85149509992 (Scopus ID)
Note

QC 20230412

Available from: 2023-04-12 Created: 2023-04-12 Last updated: 2025-02-20Bibliographically approved
Li, H., Lu, Z., Hao, M.-S., Kvammen, A., Inman, A. R., Srivastava, V., . . . McKee, L. S. (2023). Family 92 carbohydrate-binding modules specific for β-1,6-glucans increase the thermostability of a bacterial chitinase. Biochimie, 212, 153-160
Open this publication in new window or tab >>Family 92 carbohydrate-binding modules specific for β-1,6-glucans increase the thermostability of a bacterial chitinase
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2023 (English)In: Biochimie, ISSN 0300-9084, E-ISSN 1638-6183, Vol. 212, p. 153-160Article in journal (Refereed) Published
Abstract [en]

In biomass-processing industries there is a need for enzymes that can withstand high temperatures. Extensive research efforts have been dedicated to finding new thermostable enzymes as well as developing new means of stabilising existing enzymes. The attachment of a stable non-catalytic domain to an enzyme can, in some instances, protect a biocatalyst from thermal denaturation. Carbohydrate-binding modules (CBMs) are non-catalytic domains typically found appended to biomass-degrading or modifying enzymes, such as glycoside hydrolases (GHs). Most often, CBMs interact with the same polysaccharide as their enzyme partners, leading to an enhanced reaction rate via the promotion of enzyme-substrate interactions. Contradictory to this general concept, we show an example of a chitin-degrading enzyme from GH family 18 that is appended to two CBM domains from family 92, both of which bind preferentially to the non-substrate polysaccharide β-1,6-glucan. During chitin hydrolysis, the CBMs do not contribute to enzyme-substrate interactions but instead confer a 10–15 °C increase in enzyme thermal stability. We propose that CBM92 domains may have a natural enzyme stabilisation role in some cases, which may be relevant to enzyme design for high-temperature applications in biorefinery.

Place, publisher, year, edition, pages
Elsevier BV, 2023
National Category
Biocatalysis and Enzyme Technology Biochemistry Molecular Biology
Identifiers
urn:nbn:se:kth:diva-326982 (URN)10.1016/j.biochi.2023.04.019 (DOI)001053569500001 ()37121306 (PubMedID)2-s2.0-85154580139 (Scopus ID)
Note

QC 20230516

Available from: 2023-05-16 Created: 2023-05-16 Last updated: 2025-02-20Bibliographically approved
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Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0003-1877-4154

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