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. 2017 Oct 11;7:12963. doi: 10.1038/s41598-017-13526-6

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© The Author(s) 2017

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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Chitin substrates are degraded by AMCase under gastrointestinal condition. Purified AMCase was incubated at 37 °C for 0, 10, 40, and 60 min under intestine-like environment in the presence of trypsin/chymotrypsin. (a) The samples were analyzed by SDS-PAGE followed by SYPRO Ruby staining. Full-length gel is shown in Supplementary Fig. S6. (b) Determination of the chitinolytic activity. A, AMCase only; T/C, trypsin/chymotrypsin only; numbers, incubation time of AMCase and trypsin/chymotrypsin in minutes. Values in (b) represent mean ± SD conducted in triplicate. Degradation products generated by incubation of (c,e) crystalline or (d,f) colloidal chitin with (c,d) purified enzyme under the intestine condition or with (e,f) the stomach extract mimicking GIT conditions were analyzed by FACE. Full-length gels are shown in Supplementary Fig. S6.