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Chondroitin Lyase from a Marine Arthrobacter sp. MAT3885 for the Production of Chondroitin Sulfate Disaccharides

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Abstract

Chondroitin sulfate (CS) saccharides from cartilage tissues have potential application in medicine or as dietary supplements due to their therapeutic bioactivities. Studies have shown that depolymerized CS saccharides may display enhanced bioactivity. The objective of this study was to isolate a CS-degrading enzyme for an efficient production of CS oligo- or disaccharides. CS-degrading bacteria from marine environments were enriched using in situ artificial support colonization containing CS from shark cartilage as substrate. Subsequently, an Arthrobacter species (strain MAT3885) efficiently degrading CS was isolated from a CS enrichment culture. The genomic DNA from strain MAT3885 was pyro-sequenced by using the 454 FLX sequencing technology. Following assembly and annotation, an orf, annotated as family 8 polysaccharide lyase genes, was identified, encoding an amino acid sequence with a similarity to CS lyases according to NCBI blastX. The gene, designated choA1, was cloned in Escherichia coli and expressed downstream of and in frame with the E. coli malE gene for obtaining a high yield of soluble recombinant protein. Applying a dual-tag system (MalE-Smt3-ChoA1), the MalE domain was separated from ChoA1 with proteolytic cleavage using Ulp1 protease. ChoA1 was defined as an AC-type enzyme as it degraded chondroitin sulfate A, C, and hyaluronic acid. The optimum activity of the enzyme was at pH 5.5–7.5 and 40 °C, running a 10-min reaction. The native enzyme was estimated to be a monomer. As the recombinant chondroitin sulfate lyase (designated as ChoA1R) degraded chondroitin sulfate efficiently compared to a benchmark enzyme, it may be used for the production of chondroitin sulfate disaccharides for the food industry or health-promoting products.

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Acknowledgments

This work was supported by a grant from the Technology Development Fund and the AVS R&D Fund of the Ministry of Fisheries and Agriculture in Iceland and the European Union, projects 265992 (Amylomics) and 311932 (SeaBioTech). We thank Solveig Olafsdottir, Brynjar Örn Ellertsson and Dr. Björn Viðar Aðalbjörnsson for technical assistance, Dr. Josef Altenbuchner for providing the expression vectors and Dr. Bryndis Björnsdottir for critical reading of the manuscript.

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  1. Matís, Vínlandsleið 12, 113, Reykjavík, Iceland

    Varsha Kale, Ólafur Friðjónsson, Jón Óskar Jónsson, Hörður G. Kristinsson & Guðmundur Ó. Hreggviðsson

  2. Faculty of Pharmaceutical Sciences, University of Iceland, Hagi, Hofsvallagata 53, 101, Reykjavík, Iceland

    Varsha Kale & Sesselja Ómarsdóttir

  3. Department of Biology, University of Iceland, Sturlugata 7, 101, Reykjavík, Iceland

    Guðmundur Ó. Hreggviðsson

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  1. Varsha Kale
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  2. Ólafur Friðjónsson
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Kale, V., Friðjónsson, Ó., Jónsson, J.Ó. et al. Chondroitin Lyase from a Marine Arthrobacter sp. MAT3885 for the Production of Chondroitin Sulfate Disaccharides. Mar Biotechnol 17, 479–492 (2015). https://doi.org/10.1007/s10126-015-9629-9

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