Enzymatic depolymerization of alginate by two novel thermostable alginate lyases from Rhodothermus marinus

Enzymatic depolymerization of alginate by two novel thermostable alginate lyases from Rhodothermus marinus

Alginate (alginic acid) is a linear polysaccharide, wherein (1→4)-linked β- D -mannuronic acid and its C5 epimer, α- L -guluronic acid, are arranged in varying sequences. Alginate lyases catalyze the depolymerization of alginate, thereby cleaving the (1→4) glycosidic linkages between the monomers by...

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Published in:Frontiers in plant science Vol. 13; p. 981602
Main Authors: Dobruchowska, Justyna M., Bjornsdottir, Bryndis, Fridjonsson, Olafur H., Altenbuchner, Josef, Watzlawick, Hildegard, Gerwig, Gerrit J., Dijkhuizen, Lubbert, Kamerling, Johannis P., Hreggvidsson, Gudmundur O.
Format: Journal Article
Language:English
Published: Frontiers Media S.A 20-09-2022
Subjects:
alginate oligosaccharides
guluronic acid
mannuronic acid
NMR spectroscopy
Plant Science
Rhodothermus marinus
thermophilic alginate lyase
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Summary:Alginate (alginic acid) is a linear polysaccharide, wherein (1→4)-linked β- D -mannuronic acid and its C5 epimer, α- L -guluronic acid, are arranged in varying sequences. Alginate lyases catalyze the depolymerization of alginate, thereby cleaving the (1→4) glycosidic linkages between the monomers by a β-elimination mechanism, to yield unsaturated 4-deoxy- L - erythro -hex-4-enopyranosyluronic acid (Δ) at the non-reducing end of resulting oligosaccharides (α- L - erythro configuration) or, depending on the enzyme, the unsaturated monosaccharide itself. In solution, the released free unsaturated monomer product is further hydrated in a spontaneous (keto-enol tautomerization) process to form two cyclic stereoisomers. In this study, two alginate lyase genes, designated alyRm3 and alyRm4 , from the marine thermophilic bacterium Rhodothermus marinus (strain MAT378), were cloned and expressed in Escherichia coli . The recombinant enzymes were characterized, and their substrate specificity and product structures determined. AlyRm3 (PL39) and AlyRm4 (PL17) are among the most thermophilic and thermostable alginate lyases described to date with temperature optimum of activity at ∼75 and 81°C, respectively. The pH optimum of activity of AlyRm3 is ∼5.5 and AlyRm4 at pH 6.5. Detailed NMR analysis of the incubation products demonstrated that AlyRm3 is an endolytic lyase, while AlyRm4 is an exolytic lyase, cleaving monomers from the non-reducing end of oligo/poly-alginates.
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Reviewed by: Rufeng Wang, Shanghai University of Traditional Chinese Medicine, China; Jun-ichi Kadokawa, Kagoshima University, Japan
This article was submitted to Marine and Freshwater Plants, a section of the journal Frontiers in Plant Science
These authors have contributed equally to this work
Edited by: Ugo Cenci, Lille University of Science and Technology, France
Present address: Lubbert Dijkhuizen, CarbExplore Research BV, Groningen, Netherlands Justyna M. Dobruchowska, Chemical Biology and Drug Discovery, Utrecht University, Utrecht, Netherlands
ISSN:1664-462X
1664-462X
DOI:10.3389/fpls.2022.981602