Structural investigation of α-l-fucosidase from the pancreas of Patiria pectinifera, based on molecular cloning

Ono, Akiko; Suzuki, Tomohiro; Gotoh, Saki; Kono, Haruka; Matsui, Masayuki; Aoki, Daisuke; Matsuda, Masaru; Kawagishi, Hirokazu; Ogata, Makoto

doi:10.1016/j.carres.2019.02.001

Cited by 8 publications

(5 citation statements)

References 30 publications

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“…Fucosidase ATCC_03833 showed the highest catalytic efficiency with a k cat of 83.6 s −1 and a K m of 179.1 µM (Table 1). These values are consistent with fucosidases belonging to GH29 subfamily A [15,17,63,64]. Fucosidases ATCC_00842 and E1_10180 also showed activity against pNP-Fuc, but their high K m suggest that pNP-Fuc is not a good ligand for these enzymes.…”

Section: R Gnavus Fucosidases From Gh29 and Gh95 Families Display Nosupporting

confidence: 78%

Fucosidases from the human gut symbiont Ruminococcus gnavus

Rebello

Crost

et al. 2020

Cell. Mol. Life Sci.

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The availability and repartition of fucosylated glycans within the gastrointestinal tract contributes to the adaptation of gut bacteria species to ecological niches. To access this source of nutrients, gut bacteria encode α-l-fucosidases (fucosidases) which catalyze the hydrolysis of terminal α-l-fucosidic linkages. We determined the substrate and linkage specificities of fucosidases from the human gut symbiont Ruminococcus gnavus. Sequence similarity network identified strain-specific fucosidases in R. gnavus ATCC 29149 and E1 strains that were further validated enzymatically against a range of defined oligosaccharides and glycoconjugates. Using a combination of glycan microarrays, mass spectrometry, isothermal titration calorimetry, crystallographic and saturation transfer difference NMR approaches, we identified a fucosidase with the capacity to recognize sialic acid-terminated fucosylated glycans (sialyl Lewis X/A epitopes) and hydrolyze α1-3/4 fucosyl linkages in these substrates without the need to remove sialic acid. Molecular dynamics simulation and docking showed that 3′-Sialyl Lewis X (sLeX) could be accommodated within the binding site of the enzyme. This specificity may contribute to the adaptation of R. gnavus strains to the infant and adult gut and has potential applications in diagnostic glycomic assays for diabetes and certain cancers.

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Section: R Gnavus Fucosidases From Gh29 and Gh95 Families Display Nosupporting

confidence: 78%

Fucosidases from the human gut symbiont Ruminococcus gnavus

Rebello

Crost

et al. 2020

Cell. Mol. Life Sci.

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“…The following supporting information can be downloaded at: https:// www.mdpi.com/article/10.3390/foods12071555/s1, Figure S1: The SDS-PAGE results of fucoidanase; Table S1: Factors of orthogonal experiment; Table S2: The results of L9 (34)…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, fucoidanase has been shown to exist in marine molluscs such as Lambis sp. [ 32 ], Littorina kurila [ 33 ], marine echinoderms, the pancreas of Patiria pectinifera [ 34 ], sea cucumber, marine urchins such as Strongylocentrotus nudus [ 35 ], marine shellfish such as Haliotus sp. [ 36 ], Patinopecten (Mizuhopecten) yessoensis [ 37 ] and marine bacteria such as Fucobacter marina SI-0098 ( Flavobacterium sp.…”

Section: Introductionmentioning

confidence: 99%

Purification and Characterization of the Enzyme Fucoidanase from Cobetia amphilecti Utilizing Fucoidan from Undaria pinnatifida

Liu

Wang

Shao³

et al. 2023

Foods

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Fucoidanase is an unstable enzyme with high specificity that requires a large about of time to screen it from microorganisms. In this study, enzymatic hydrolysis was used to produce low-molecular-weight fucoidan from microorganisms via the degradation of high-molecular-weight fucoidan without damage to the sulfate esterification structure of oligosaccharide. The microbial strain HN-25 was isolated from sea mud and was made to undergo mutagenicity under ultraviolet light. Fucoidanase was extracted via ultrasonication and its enzymatic activity was improved via optimization of the ultrasonic conditions. The enzymatic properties and degradation efficiency of fucoidanase were characterized. The microbial strain HN-25 is a Gram-negative aerobic and rod-shaped-cell bacterium, and therefore was identified as Cobetia amphilecti via 16s rDNA. The results proved that fucoidanase is a hydrolytic enzyme with a molecular weight of 35 kDa and with high activity and stability at 30 °C and pH 8.0. The activity of fucoidanase was significantly enhanced by sodium and calcium ions and inhibited by a copper ion and ethylenediaminetetraacetate (EDTA). There was a significant decrease in the molecular weight of fucoidan after enzymatic hydrolysis. The low-molecular-weight fuicodan was divided into four fractions, mainly concentrated at F3 (20~10 kDa) and F4 (≤6 kDa). These consequences suggest that fucoidanase obtained from Cobetia amphilecti is stable and efficient and could be a good tool in the production of bioactive compounds.

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“…Most characterized GH29 fucosidases are of terrestrial origin, while only a few are from marine organisms (Tarling et al 2003;Dong et al 2017;Ono et al 2019;Hong et al 2021). Terrestrial and marine glycans have distinctly different compositions, and especially, algal cell walls contain polysaccharides (e.g., fucoidan), which are not found in organisms living on land (Popper et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Characterization of five marine family 29 glycoside hydrolases reveals an α-L-fucosidase targeting specifically Fuc(α1,4)GlcNAc

et al. 2022

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l-Fucose is the most widely distributed l-hexose in marine and terrestrial environments, and presents a variety of functional roles. l-Fucose is the major monosaccharide in the polysaccharide fucoidan from cell walls of brown algae, and is found in human milk oligosaccharides and the Lewis blood group system, where it is important in cell signaling and immune response stimulation. Removal of fucose from these biomolecules is catalyzed by fucosidases belonging to different carbohydrate-active enzyme (CAZy) families. Fucosidases of glycoside hydrolase family 29 (GH29) release α-l-fucose from non-reducing ends of glycans and display activities targeting different substrate compositions and linkage types. While several GH29 fucosidases from terrestrial environments have been characterized, much less is known about marine members of GH29 and their substrate specificities, as only four marine GH29 enzymes were previously characterized. Here, five GH29 fucosidases originating from an uncultured fucoidan-degrading marine bacterium (Paraglaciecola sp.) were cloned and produced recombinantly in E. coli. All five enzymes (Fp231, Fp239, Fp240, Fp251, Fp284) hydrolyzed the synthetic substrate CNP-α-l-fucose. Assayed against up to 17 fucose-containing oligosaccharides, Fp239 showed activity against the Lewis Y antigen, 2′- and 3-fucosyllactose while Fp284 degraded 2′-fucosyllactose and Fuc(α1,6)GlcNAc. Furthermore, Fp231 displayed strict specificity against Fuc(α1,4)GlcNAc, a previously unreported specificity in GH29. Fp231 is a monomeric enzyme with pH and temperature optima at pH 5.6–6.0 and 25 °C, hydrolyzing Fuc(α1,4)GlcNAc with kcat = 1.3 s−1 and Km = 660 μM. Altogether, the findings extend our knowledge about GH29 family members from the marine environment, which are so far largely unexplored.

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Structural investigation of α-l-fucosidase from the pancreas of Patiria pectinifera, based on molecular cloning

Cited by 8 publications

References 30 publications

Fucosidases from the human gut symbiont Ruminococcus gnavus

Fucosidases from the human gut symbiont Ruminococcus gnavus

Purification and Characterization of the Enzyme Fucoidanase from Cobetia amphilecti Utilizing Fucoidan from Undaria pinnatifida

Characterization of five marine family 29 glycoside hydrolases reveals an α-L-fucosidase targeting specifically Fuc(α1,4)GlcNAc

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