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1
Antimicrobial Properties and Cytotoxicity of Sulfated (1,3)-β-D-Glucan from the Mycelium of the Mushroom Ganoderma lucidum

( Wan Abd Al Qadr Imad Wan Mohtar ),( Louise Young ),( Grainne M. Abbott ),( Carol Clements ),( Linda M. Harvey ),( Brian Mcneil ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.6
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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.
2
An LCMS method for the assay of melittin in cosmetic formulations containing bee venom.

Tusiimire, Jonans,Wallace, Jennifer,Dufton, Mark,Parkinson, John,Clements, Carol J,Young, Louise,Park, Jin Kyu,Jeon, Jong Woon,Watson, David G Springer-Verlag 2015 Analytical and Bioanalytical Chemistry Vol.407 No.13
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<P>There is a growing interest in the potential of bee venom in cosmetics as a rejuvenating agent. Products currently on the market do not specify exactly their content of bee venom (BV). Therefore, we developed a method for the detection and quantification of melittin, as a marker of bee venom content, in selected commercial creams which contained BV according to their marketing claims, in order to gauge the relative quality of such formulations. A quantitative method was achieved following a rigorous extraction procedure involving sonication, liquid-liquid extraction and solid phase extraction since carryover of excipients was found to cause a rapid deterioration in the chromatographic performance. The method employed a standard additions approach using, as spiking standard, purified melittin isolated from bee venom and standardised by quantitative NMR. The aqueous extracts of the spiked creams were analysed by reversed phase LCMS on an LTQ Orbitrap mass spectrometer. The purity of the melittin spiking standard was determined to be 96.0%. The lowest measured mean melittin content in the creams was 3.19 ppm (1.58 ppm 95% CI) while the highest was 37.21 ppm (2.01 ppm 95% CI). The method showed adequate linearity (R (2)??0.98) and a recovery of 87.7-102.2% from a spiked blank cream. An assay precision of <20% RSD was achieved for all but one sample where the RSD value was 27.5%. The method was sensitive enough for use in routine assay of BV-containing cosmetic creams. Differences in the melittin content of the commercial products assayed were nearly tenfold.</P>