An exocellular β-(1→6)-D-glucan (lasiodiplodan) produced by a strain of Lasiodiplodia theobromae (MMLR) grown on sucrose was derivatized by sulfonation to promote anticoagulant activity. The structural features of the sulfonated β-(1→6)-D-glucan were investigated by UV-vis, FT-IR and (13)C NMR spectroscopy, and the anticoagulant activity was investigated by the classical coagulation assays APTT, PT and TT using heparin as standard. The content of sulfur and degree of substitution of the sulfonated glucan was 11.73% and 0.95, respectively. UV spectroscopy showed a band at 261 nm due to the unsaturated bond formed in the sulfonation reaction. Results of FT-IR and (13)C NMR indicated that sulfonyl groups were inserted on the polysaccharide. The sulfonated β-(1→6)-D-glucan presented anticoagulant activity as demonstrated by the increase in dose dependence of APTT and TT, and these actions most likely occurred because of the inserted sulfonate groups on the polysaccharide. The lasiodiplodan did not inhibit the coagulation tests.
Microbial hyaluronic acid (HA) production has been preferred rather than extraction from animal tissue for medical and cosmetic applications. In this context, to obtain an economically competitive HA production by Streptococcus zooepidemicus, culture conditions were studied to improve the polymer production in sugarcane molasses. The highest HA production by S. zooepidemicus ATCC 39920 achieved was 2.825 g. L in a 4.5 L bioreactor with controlled pH (8.0) and medium containing molasses (85.35 g.L total sugar) pretreated with activated charcoal and yeast extract (50 g.L). The HA produced exhibited a high molecular weight of 1.35 × 10 kDa and the DPPH radical scavenging activity of the polymer at 1 g.L was 41 %. The FTIR and UV-Vis spectra showed no substantial differences in the spectral pattern between produced and standard HA. This study is a promising strategy for sugarcane molasses application by producing high value-added products such as hyaluronic acid.
Sugarcane bagasse was used as substrate for xylanase production by means of a strain of Trichoderma harzianum Rifai isolated from decaying Aspidosperma sp. (peroba) wood. The bagasse was washed, dried, milled and wetted with minimal salts medium and the cultures grown at 28 ± 2ºC for 7 days. Two extraction methods were tested for enzyme recovery: (A) Tween 80, 0.1% (v/v), in physiological saline, and (B) 50mM sodium acetate buffer, pH 5.0, under agitation (180rpm) for 15, 30 and 60min. After a single extraction, both extraction methods recovered an average of 15U/ml of xylanase activity, independent on the time of shaking. A second and third extraction recovered 10.4 and 6.6U/ml xylanase, respectively. The effect of volume size for extraction, and sugarcane bagasse concentration, on xylanase production were also investigated. The growth profile of Trichoderma harzianum was followed over 20 days on 14% (w/v) bagasse, and highest xylanase activity (288U/ml) appeared on the seventh day. The enzymatic extract after precipitation with ammonium sulphate was submitted to electrophoresis on polyacrylamide gels and showed 4 protein-staining bands, one of which exhibited xylanase activity.
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