Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4

Liu, Zhanliang; Tuveng, Tina Rise; Wittrup_Agger, Jane; Westereng, Bjørge; Mathiesen, Geir; Horn, Svein Jarle; Vaaje‐Kolstad, Gustav; Aalten, Dmf van; Eijsink, Vincentius Gerardus Henricus

doi:10.1038/s41598-017-02043-1

Cited by 56 publications

(66 citation statements)

References 52 publications

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“…Diatom CDAs show higher activity against chitin oligomers than polymers (DA20 and DA60). This is contrary to the activity of CDAs from fungi, which show higher activity on DA60 than on chitin pentamers (Cord‐Landwehr et al .,; Liu et al ., ). TpCDA was more active on highly acetylated chitosan polymers DA60 than on DA20, which is consistent with fungal CDA from Pestalotiopsis sp.…”

Section: Discussionmentioning

confidence: 97%

Comparative characterization of putative chitin deacetylases from Phaeodactylum tricornutum and Thalassiosira pseudonana highlights the potential for distinct chitin‐based metabolic processes in diatoms

Shao

Thomas

Hembach³

et al. 2018

New Phytologist

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Summary Chitin is generally considered to be present in centric diatoms but not in pennate species. Many aspects of chitin biosynthetic pathways have not been explored in diatoms. We retrieved chitin metabolic genes from pennate (Phaeodactylum tricornutum) and centric (Thalassiosira pseudonana) diatom genomes. Chitin deacetylase (CDA) genes from each genome (PtCDA and TpCDA) were overexpressed in P. tricornutum. We performed comparative analysis of their sequence structure, phylogeny, transcriptional profiles, localization and enzymatic activities. The chitin relevant proteins show complex subcellular compartmentation. PtCDA was likely acquired by horizontal gene transfer from prokaryotes, whereas TpCDA has closer relationships with sequences in Opisthokonta. Using transgenic P. tricornutum lines expressing CDA‐green fluorescent protein (GFP) fusion proteins, PtCDA predominantly localizes to Golgi apparatus whereas TpCDA localizes to endoplasmic reticulum/chloroplast endoplasmic reticulum membrane. CDA‐GFP overexpression upregulated the transcription of chitin synthases and potentially enhanced the ability of chitin synthesis. Although both CDAs are active on GlcNAc5, TpCDA is more active on the highly acetylated chitin polymer DA60. We have addressed the ambiguous characters of CDAs from P. tricornutum and T. pseudonana. Differences in localization, evolution, expression and activities provide explanations underlying the greater potential of centric diatoms for chitin biosynthesis. This study paves the way for in vitro applications of novel CDAs.

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Section: Discussionmentioning

confidence: 97%

Comparative characterization of putative chitin deacetylases from Phaeodactylum tricornutum and Thalassiosira pseudonana highlights the potential for distinct chitin‐based metabolic processes in diatoms

Shao

Thomas

Hembach³

et al. 2018

New Phytologist

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“…Current CAZyme research has focused in the isolation and characterization of novel enzymes (Attia et al 2016;Iacono et al 2019;Liu et al 2017;Munoz-Munoz et al 2017). While these studies have elucidated the enzymatic mechanisms of several CAZymes using artificial substrates, there are fewer studies that employ environmentally relevant substrates because of the practical difficulties of performing enzymatic assays using these materials.…”

Section: Discussionmentioning

confidence: 99%

High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates

Monge¹,

Levi²,

Forbin³

et al. 2020

Appl Microbiol Biotechnol

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Carbohydrate degradation by microbes plays an important role in global nutrient cycling, human nutrition, and biotechnological applications. Studies that focus on the degradation of complex recalcitrant polysaccharides are challenging because of the insolubility of these substrates as found in their natural contexts. Specifically, current methods to examine carbohydrate-based biomass degradation using bacterial strains or purified enzymes are not compatible with high-throughput screening using complex insoluble materials. In this report, we developed a small 3D printed filter device that fits inside a microplate well that allows for the free movement of bacterial cells, media, and enzymes while containing insoluble biomass. These devices do not interfere with standard microplate readers and can be used for both short-(24-48 h) and long-duration (> 100 h) experiments using complex insoluble substrates. These devices were used to quantitatively screen in a high-throughput manner environmental isolates for their ability to grow using lignocellulose or rice grains as a sole nutrient source. Additionally, we determined that the microplate-based containment devices are compatible with existing enzymatic assays to measure activity against insoluble biomass. Overall, these microplate containment devices provide a platform to study the degradation of complex insoluble materials in a high-throughput manner and have the potential to help uncover ecologically important aspects of bacterial metabolism as well as to accelerate biotechnological innovation.

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“…Evolutionary history was inferred using the maximum likelihood method based on a JTT matrix-based model 52 . The amino acid sequences used in this tree were reported in [37][38][39][40][41][42][43][44][45][46][47] . Clustal-X2 (Version 1.83) was used for multiple-sequence alignment.…”

Section: Methodsmentioning

confidence: 99%

Isolation, characterisation, and genome sequencing of Rhodococcus equi: a novel strain producing chitin deacetylase

Ma¹,

Gao

Bi³

et al. 2020

Sci Rep

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chitin deacetylase (cDA) can hydrolyse the acetamido group of chitin polymers to produce chitosans, which are used in various fields including the biomedical and pharmaceutical industries, food production, agriculture, and water treatment. cDA represents a more environmentally-friendly and easier to control alternative to the chemical methods currently utilised to produce chitosans from chitin; however, the majority of identified CDAs display activity toward low-molecular-weight oligomers and are essentially inactive toward polymeric chitin or chitosans. therefore, it is important to identify novel cDAs with activity toward polymeric chitin and chitosans. in this study, we isolated the bacterium Rhodococcus equi F6 from a soil sample and showed that it expresses a novel CDA (ReCDA), whose activity toward 4-nitroacetanilide reached 19.20 U/mL/h during fermentation and was able to deacetylate polymeric chitin, colloidal chitin, glycol-chitin, and chitosan. Whole genome sequencing revealed that RecDA is unique to the R. equi F6 genome, while phylogenetic analysis indicated that RecDA is evolutionarily distant from other cDAs. in conclusion, RecDA isolated from the R. equi F6 strain expands the known repertoire of CDAs and could be used to deacetylate polymeric chitosans and chitin in industrial applications.Chitin is the second most abundant biopolymer after cellulose and is mainly obtained as a waste product of the seafood industry at a relatively low cost 1 . The chitin derivative, chitosan, is a linear polysaccharide comprised of β-(1→4)-linked glucosamine and N-acetyl glucosamine units which are randomly arranged within the chitosan polysaccharide chain 2 . Due to its ability to dissolve in dilute acids, chitosan is more useful than its crystalline precursor chitin in various industrial applications 3 , including the biomedical and pharmaceutical industries, food production, agriculture, and water treatment 4 .Chitosan occurs naturally and is mainly found in the cell walls of certain fungi, the exoskeletons of certain insects (such as the abdominal wall of termite queens), and in some yeasts 5,6 . Although chitosan can be extracted from fungal sources 7 , the method is commercially inapplicable as it provides too low a yield at too high a cost. Therefore, chitosan is still obtained by treating marine-derived chitin with thermo-alkaline 8 , a method that is inexpensive and results in high yields but is environmentally unsafe and difficult to control, producing a heterogeneous range of products depending on the degree of deacetylation 9 . The enzymatic production of chitosan using microbial chitin deacetylase (CDA; EC 3.5.1.41), which can hydrolyse the acetamido group in chitin polymers to produce chitosans 10 , is an environmentally-friendly process that is easy to control and results in a high yield of homogeneous end products 11 .CDAs have been identified in marine and soil bacteria, several fungi, a few insects, and at least one virus 12,13 . Fungal CDAs exist mostly as N-glycosylated (20-70%) glycoproteins, w...

show abstract

Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4

Cited by 56 publications

References 52 publications

Comparative characterization of putative chitin deacetylases from Phaeodactylum tricornutum and Thalassiosira pseudonana highlights the potential for distinct chitin‐based metabolic processes in diatoms

Comparative characterization of putative chitin deacetylases from Phaeodactylum tricornutum and Thalassiosira pseudonana highlights the potential for distinct chitin‐based metabolic processes in diatoms

High-throughput screening of environmental polysaccharide-degrading bacteria using biomass containment and complex insoluble substrates

Isolation, characterisation, and genome sequencing of Rhodococcus equi: a novel strain producing chitin deacetylase

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