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. 2018 Oct 2;8:14713. doi: 10.1038/s41598-018-32922-0

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Purification of the Cdf79930 and its activity against ulvan. (A) Purification of Cdf79930 by size exclusion chromatography and SDS-PAGE analysis of the purified fractions: lane 1, molecular weight standard; lane 2, Cdf79930 fraction after immobilised metal affinity chromatography; lane 3, fraction 1 after size exclusion chromatography; and lane 4, fraction 2 after size exclusion chromatography. The picture of the SDS-PAGE gel was cropped; full gel picture is added as Fig. S7 in the Supplementary Information. (B) Michaelis-Menten plot of Cdf79930 with ulvan extracted from U. amoricana as substrate. Each point represents the average value of triplicate measurements, and the error bar indicate the standard error. The fitted K_m and V_max values are 0.77 mg ml⁻¹ and 540 µg of reducing end equivalents of glucose ml⁻¹ min⁻¹ mg protein⁻¹, respectively. (C) Substrate specificity of the novel ulvan lyase. Cdf79930 was incubated with 200 µg of each substrate and incubated for 60 minutes at 37 °C. Cdf79930 depolymerised both chondroitin sulphate and heparan sulphate. (D) Separation of end-products from the Cdf79930 action on ulvan using size exclusion chromatography.