Marques, LuísKarmali, Amin2026-05-042026-05-042025-12-07Marques, L., & Karmali, A. (2025). Experimental planning for production of β-D-glucan: purification and fluorescence properties from basidiomycete strains. Separations, 12(12), 1-17. https://doi.org/10.3390/separations12120336http://hdl.handle.net/10400.21/22850This research was funded by the European Investment Funds by FEDER/COMPETE/POCI–Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT—Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2019.first_pagesettingsOrder Article Reprints Open AccessArticle Experimental Planning for Production of β-D-Glucan: Purification and Fluorescence Properties from Basidiomycete Strains by Luís Marques 1 andAmin Karmali 1,2,* 1 Chemical Engineering and Biotechnology Research Center and Department of Chemical Engineering of Instituto Superior de Engenharia de Lisboa, Rua Conselheiro Emídio Navarro No.1, 1950-072 Lisboa, Portugal 2 Centre for the Research and Technology for Agro-Environmental and Biological Sciences, Universidade de Trás-os-Montes e Alto Douro Quinta dos Prados, Apartado 1013, 5001-801 Vila Real, Portugal * Author to whom correspondence should be addressed. Separations 2025, 12(12), 336; https://doi.org/10.3390/separations12120336 Submission received: 1 November 2025 / Revised: 28 November 2025 / Accepted: 1 December 2025 / Published: 7 December 2025 Downloadkeyboard_arrow_down Browse Figures Versions Notes Abstract Fruit and coffee industries are responsible for huge quantities of agro-industrial wastes which is of great environmental and public health concern. Therefore, the aim of this work involves the use of such wastes for the production of β-D-glucan from basidiomycete strains which are powerful biological response modifiers in several clinical disorders. Experimental planning for optimization of several parameters was carried out by a full factorial of two levels of three factors for production of beta-glucans and basidiomycete species, where waste concentration and interaction between species and agro-industrial waste were the most important factors. The best conditions involved a basidiomycete strain of Lentinula edodes in a culture medium containing 400 g/L of waste coffee grounds which revealed the production of extracellular β-glucans (141.16 mg/L) at the 3rd day of fermentation. Intrinsic fluorescence properties of mushroom β-D-glucan were investigated by fluorescence spectroscopy as well as a fluorescence microtiter plate reader exhibiting emission peaks at 492 and 528 nm. Differential chromatographic behavior of β-D-glucan was investigated by immobilized metal affinity chromatography (IMAC) using epoxy-activated Sepharose 6B containing different chelating agents, spacer arms, and metal ions. One-step purification of β-D-glucan was devised using a column of epoxy-activated Sepharose 6B-IDA-Cu (II). FTIR analysis of several β-D-glucans from the chromatographic fractions was carried out to investigate their structural properties.engFruit and coffee industry wastesExperimental planningExtracellular β-D-glucan from basidiomycete mushroomsImmobilized metal affinity chromatographyIMACIntrinsic fluorescence of β-D-glucanFTIRUID/AGR/04033/2019research article10.3390/separations121203362297-8739