CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136
Gelidibacter salicanalis strains remain poorly understood in terms of their genomic attributes and are rarely reported for the degradation of polysaccharides within the niche. Gelidibacter salicanalis PAMC21136, an Antarctic isolate, was sequenced and functionally annotated. 251 genes were classifie...
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Elsevier
2025-06-01
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| Series: | Carbohydrate Polymer Technologies and Applications |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666893925001124 |
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| author | Lakshan Paudel Bashu Dev Pardhe So-Ra Han Jun Hyuck Lee Tae-Jin Oh |
| author_facet | Lakshan Paudel Bashu Dev Pardhe So-Ra Han Jun Hyuck Lee Tae-Jin Oh |
| author_sort | Lakshan Paudel |
| collection | DOAJ |
| description | Gelidibacter salicanalis strains remain poorly understood in terms of their genomic attributes and are rarely reported for the degradation of polysaccharides within the niche. Gelidibacter salicanalis PAMC21136, an Antarctic isolate, was sequenced and functionally annotated. 251 genes were classified into the CAZyme families, which includes the highest number of GH family, highlighting peculiarity towards various macro biopolymers. Further, based on domain architecture and multiple sequence alignment, gene belonging to the family18 glycoside hydrolase was identified as chitinase. The gene encoding chitinase (MBJ7879808.1) was successfully cloned and expressed in Escherichia coli BL21 cells. MBJ7879808.1 demonstrated activity towards colloidal chitin but there was no detectable activity towards microcrystalline crab shell chitin. Enzyme exhibited an exo-pattern of hydrolysis on both colloidal chitin, and shorter chain GlcNAc oligomers namely [(GlcNAc)3, (GlcNAc)4, (GlcNAc)5, and (GlcNAc)6] producing GlcNAc as a single final product. Biochemical characterization revealed an optimum activity at 25 °C and pH 7.0. Mn²⁺ ions significantly enhanced the activity by 1.7-fold. Turnover number (Kcat) was 8.6 min−1 and catalytic efficiency (Kcat/Km) was 2.0ml/mg/min towards the substrate colloidal chitin. These findings expanded the polysaccharide degradation capability of Gelidibacter salicanalis PAMC21136 and highlight the biotechnological potential of the GH18 enzyme towards production of chitooligomers. |
| format | Article |
| id | doaj-art-e85338fc6e7c403ea2fb2909fbddaabf |
| institution | DOAJ |
| issn | 2666-8939 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
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| series | Carbohydrate Polymer Technologies and Applications |
| spelling | doaj-art-e85338fc6e7c403ea2fb2909fbddaabf2025-08-20T02:39:35ZengElsevierCarbohydrate Polymer Technologies and Applications2666-89392025-06-011010077310.1016/j.carpta.2025.100773CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136Lakshan Paudel0Bashu Dev Pardhe1So-Ra Han2Jun Hyuck Lee3Tae-Jin Oh4Department of Life Science and Biochemical Engineering, Graduate School, SunMoon University, Asan 31460, South KoreaDepartment of Life Science and Biochemical Engineering, Graduate School, SunMoon University, Asan 31460, South KoreaGenome-based BioIT Convergence Institute, Asan 31460, South Korea; Bio Big Data-based Chungnam Smart Clean Research Leader Training Program, SunMoon University, Asan 31460, South KoreaDivision of Life Sciences, Korea Polar Research Institute, Incheon 21990, South KoreaDepartment of Life Science and Biochemical Engineering, Graduate School, SunMoon University, Asan 31460, South Korea; Genome-based BioIT Convergence Institute, Asan 31460, South Korea; Bio Big Data-based Chungnam Smart Clean Research Leader Training Program, SunMoon University, Asan 31460, South Korea; Department of Pharmaceutical Engineering and Biotechnology, SunMoon University, Asan 31460, South Korea; Corresponding author.Gelidibacter salicanalis strains remain poorly understood in terms of their genomic attributes and are rarely reported for the degradation of polysaccharides within the niche. Gelidibacter salicanalis PAMC21136, an Antarctic isolate, was sequenced and functionally annotated. 251 genes were classified into the CAZyme families, which includes the highest number of GH family, highlighting peculiarity towards various macro biopolymers. Further, based on domain architecture and multiple sequence alignment, gene belonging to the family18 glycoside hydrolase was identified as chitinase. The gene encoding chitinase (MBJ7879808.1) was successfully cloned and expressed in Escherichia coli BL21 cells. MBJ7879808.1 demonstrated activity towards colloidal chitin but there was no detectable activity towards microcrystalline crab shell chitin. Enzyme exhibited an exo-pattern of hydrolysis on both colloidal chitin, and shorter chain GlcNAc oligomers namely [(GlcNAc)3, (GlcNAc)4, (GlcNAc)5, and (GlcNAc)6] producing GlcNAc as a single final product. Biochemical characterization revealed an optimum activity at 25 °C and pH 7.0. Mn²⁺ ions significantly enhanced the activity by 1.7-fold. Turnover number (Kcat) was 8.6 min−1 and catalytic efficiency (Kcat/Km) was 2.0ml/mg/min towards the substrate colloidal chitin. These findings expanded the polysaccharide degradation capability of Gelidibacter salicanalis PAMC21136 and highlight the biotechnological potential of the GH18 enzyme towards production of chitooligomers.http://www.sciencedirect.com/science/article/pii/S2666893925001124Antarctic bacteriaExo-chitinaseCAZymeGH18ChitooligomersN-acetyl D-glucosamine |
| spellingShingle | Lakshan Paudel Bashu Dev Pardhe So-Ra Han Jun Hyuck Lee Tae-Jin Oh CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136 Carbohydrate Polymer Technologies and Applications Antarctic bacteria Exo-chitinase CAZyme GH18 Chitooligomers N-acetyl D-glucosamine |
| title | CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136 |
| title_full | CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136 |
| title_fullStr | CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136 |
| title_full_unstemmed | CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136 |
| title_short | CAZyme analysis and functional characterization of a new GH18 chitin hydrolase from Gelidibacter salicanalis PAMC21136 |
| title_sort | cazyme analysis and functional characterization of a new gh18 chitin hydrolase from gelidibacter salicanalis pamc21136 |
| topic | Antarctic bacteria Exo-chitinase CAZyme GH18 Chitooligomers N-acetyl D-glucosamine |
| url | http://www.sciencedirect.com/science/article/pii/S2666893925001124 |
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