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. 2019 Nov 22;9:17400. doi: 10.1038/s41598-019-53726-w

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© The Author(s) 2019

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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The heterologous BuGH16 agarase is enzymatically functional in engineered B. theta strains. (a) Schematic model of BuGH16 endo-agarase activity agarose to generate neoagarooligosaccharides (NAOS). NAOS is a repeating disaccharide of l-3,6-anhydro-galactose (L-3,6) and d-galactose, with L-3,6 at the non-reducing end. (b) pINT integrated hexa-histidine tagged BuGH16 (black arrow) detected by Western blot from induced SusR^DX strains. (c) Thin-layer chromatography of culture supernatant from induced engineered strains incubated with agarose shows the presence of NAOS (black bar) generated from agarose by active BuGH16.