Wan-Mohtar, Wan Abd Al Qadr Imad
(Fermentation Centre, Strathclyde Institute of Pharmacy and Biomedical Sciences (SIPBS), Strathclyde University)
,
Young, Louise
(Strathclyde Institute for Drug Research (SIDR), SIPBS, Strathclyde University)
,
Abbott, Grainne M.
(Strathclyde Institute for Drug Research (SIDR), SIPBS, Strathclyde University)
,
Clements, Carol
(Strathclyde Institute for Drug Research (SIDR), SIPBS, Strathclyde University)
,
Harvey, Linda M.
(Fermentation Centre, Strathclyde Institute of Pharmacy and Biomedical Sciences (SIPBS), Strathclyde University)
,
McNeil, Brian
(Fermentation Centre, Strathclyde Institute of Pharmacy and Biomedical Sciences (SIPBS), Strathclyde University)
Ganoderma lucidum BCCM 31549 has a long established role for its therapeutic activities. In this context, much interest has focused on the possible functions of the (1,3)-β-D-glucan (G) produced by these cultures in a stirred-tank bioreactor and extracted from their underutilized mycelium. In the ex...
Ganoderma lucidum BCCM 31549 has a long established role for its therapeutic activities. In this context, much interest has focused on the possible functions of the (1,3)-β-D-glucan (G) produced by these cultures in a stirred-tank bioreactor and extracted from their underutilized mycelium. In the existing study, we report on the systematic production of G, and its sulfated derivative (GS). The aim of this study was to investigate G and its GS from G. lucidum in terms of their antibacterial properties and cytotoxicity spectrum against human prostate cells (PN2TA) and human caucasian histiocytic lymphoma cells (U937). 1H NMR for both G and GS compounds showed β-glycosidic linkages and structural similarities when compared with two standards (laminarin and fucoidan). The existence of characteristic absorptions at 1,170 and 867 cm-1 in the FTIR (Fourier Transform Infrared Spectroscopy) for GS demonstrated the successful sulfation of G. Only GS exhibited antimicrobial activity against a varied range of test bacteria of relevance to foodstuffs and human health. Moreover, both G and GS did not show any cytotoxic effects on PN2TA cells, thus helping demonstrate the safety of these polymers. Moreover, GS showed 40% antiproliferation against cancerous U937 cells at the low concentration (60 μg/ ml) applied in this study compared with G (10%). Together, this demonstrates that sulfation clearly improved the solubility and therapeutic activities of G. The water-soluble GS demonstrates the potential multifunctional effects of these materials in foodstuffs.
Ganoderma lucidum BCCM 31549 has a long established role for its therapeutic activities. In this context, much interest has focused on the possible functions of the (1,3)-β-D-glucan (G) produced by these cultures in a stirred-tank bioreactor and extracted from their underutilized mycelium. In the existing study, we report on the systematic production of G, and its sulfated derivative (GS). The aim of this study was to investigate G and its GS from G. lucidum in terms of their antibacterial properties and cytotoxicity spectrum against human prostate cells (PN2TA) and human caucasian histiocytic lymphoma cells (U937). 1H NMR for both G and GS compounds showed β-glycosidic linkages and structural similarities when compared with two standards (laminarin and fucoidan). The existence of characteristic absorptions at 1,170 and 867 cm-1 in the FTIR (Fourier Transform Infrared Spectroscopy) for GS demonstrated the successful sulfation of G. Only GS exhibited antimicrobial activity against a varied range of test bacteria of relevance to foodstuffs and human health. Moreover, both G and GS did not show any cytotoxic effects on PN2TA cells, thus helping demonstrate the safety of these polymers. Moreover, GS showed 40% antiproliferation against cancerous U937 cells at the low concentration (60 μg/ ml) applied in this study compared with G (10%). Together, this demonstrates that sulfation clearly improved the solubility and therapeutic activities of G. The water-soluble GS demonstrates the potential multifunctional effects of these materials in foodstuffs.
In this study, a series of dose-response assays were implemented to define the cytotoxic reactions in PN2TA. Once the cells were exposed to different concentrations (3, 5, 30, 50, 300, and 500 μg/ml) of GS and G for 24 h, the alamarBlue reagent assay displayed no loss of cell viability.
대상 데이터
Gentamicin susceptibility test discs (30 μg of concentration) were supplied by Thermo Scientific Oxoid (Fisher Scientific, UK). In this experiment, human caucasian histiocytic lymphoma (U937) and human prostate normal cells (PNT2A) were obtained from the European Collection of Cell Cultures, supplied by Sigma-Aldrich, (UK). DMEM and TrypLE Express were provided by Gibco (Life Technologies, UK).
데이터처리
All analyses were carried out in triplicate, and the respective mean ± SD was determined using the software GraphPad Prism 5 (ver. 5.
이론/모형
The method of the Strathclyde Institute of Drug Research was used for the antimicrobial test on the bacteria Klebsiella pneumoniae ATCC 13883 and Mycobacterium marinum ATCC BAA 535, using 96-well microtiter plates [4,18]. These tests were in triplicate, and the GS was supplied at 10mg/ml.
성능/효과
The introduction of the sulfate group has several purposes. Based on the present study’s findings, the aqueous solubility of the extracted G from the fermenter was as poor as that of G prepared from other procedures. Astonishingly, this was mentioned in the literature that G was less suitable for medicinal applications [13].
Elemental analysis was accomplished to attain the composition of the GS and, therefore, its degree of sulfation. Basic examination of lyophilized GS gave a composition of (w/w) 24.5% C, 5.72% H, 49.92% O, 9.85% S, and 10.01% N (Table 2). When compared with standard fucoidan, GS had the same C and H values but was slightly different in H, O, and S.
The inhibition zone diameters increased with increasing GS prepared concentrations (Table 3). These reactions displayed that the antimicrobial effect of GS was dose-dependent and that the gentamicin positive control was clearly effective against all the test bacteria.
후속연구
The mechanisms of sulfation on the structure G were proposed for these positive reactions by GS. Consequently, it is essential to further studies in order investigate glucan structure-activity relationships, which might deliver a detailed foundation for their development and improvement.
In relation to this, the GS might have a role as a natural additive in many foods, with multi functional benefits (preservative, antiproliferative, immune stimulation). Further examination of these functions for such polymers and their derivatives will be required.
Therefore, the biochemical aspects and mechanism of the antiproliferative reactions stimulated by GS from mycelium of G. lucidum is still not fully understood and requests further study.
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