N acetylglucosamine

All chemicals used in the present study were suitable for microbial culture or of analytical grade. Glucose, fructose, sucrose, galactose, lactose, glycerol, N-acetylglucosamine, and xylose were purchased from Sigma-Aldrich (St. Louis, MO, USA). Other chemicals used in the growth media were also purchased from Sigma-Aldrich or BD Difco (Franklin Lakes, NJ, USA). GC-MS authentic 3-hydroxybutyrate (3HB), 3-hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3HO), 3-hydroxydecanoate (3HD), 3-hydroxydodecanoate (3HdD), and 3-hydroxytetradecanoate (3HtD) were also purchased from Sigma-Aldrich.

C. antiquata was purchased from ZhanJiang, China. Mammalian heparin (HP), chondroitin sulfate (CS), dermatin sulfate (DS), and plasminogen were obtained from the National Institutes for Food and Drug Control (China). Alkaline protease 2709 (1.2 × 105 U/g), papain (800 U/mg), rabbit plasma, thrombin (300 U/mg), urokinase (50 U/mg), and bovine plasma fibrinogen were obtained from Yuanye Biotechnology Co. Ltd. (Shanghai, China). Activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) kits were purchased from Sun Biotech Co. Ltd. (Shanghai, China). AMBERLITE FPA98 CI macroporous anion exchange resin was purchased from Rohm and Haas (Philadelphia, PA). Standard monosaccharide references (mannose, rhamnose, glucosamine, glucuronic acid, iduronic acid, N-acetylglucosamine, glucose, galactose, arabinose, and fucose) were obtained from Sigma-Aldrich (Darmstadt, Germany). Different molecular weights of heparin standard were purchased from Adhoc International Technology Co., Ltd. (Beijing, China). All other chemicals and solvents used were of analytical grade or HPLC-grade, unless otherwise specified.

Commercial samples of COS used in this study were obtained from Qingdao (Shangdong, China) and had average molecular weights of 1000 Da and 3000 Da, respectively. The DD of the commercial COS were >90% in glucosamine hydrochloride. The N-acetylglucosamine used in this study was purchased from Sigma Aldrich (St. Louis, MO, USA) and had a purity greater than 99%. The deuterium oxide used had an isotopic purity of 99.8%, and was obtained from Shanghai Macklin Biochemical Co., Ltd. (Tianjin, China). Bromocresol green and methyl orange were purchased from Tianjin Damao Chemical Reagent Factory (Tianjin, China) and methyl orange from Tianjin Zhiyuan Chemical Reagent Co., Ltd. (Tianjin, China), respectively. Sodium hydroxide pellets (NaOH), hydrochloric acid (HCl), and potassium hydroxide pellets were obtained from Tianjin Kermel Chemical Reagent Co., Ltd. (Tianjin, China). All other reagents and solvents were of analytical grade and used without further purification. All water used in the extraction and analysis was distilled and deionized.

To determine the effect of the essential oils and their major compounds on the yeast-mycelium transition, cell suspensions of C. albicans ATCC 10231, from overnight cultures on SDA, were prepared in NYP medium [N-acetylglucosamine (Sigma; 10⁻³ mol/L), Yeast Nitrogen Base (Difco; 3.35 g/L), proline (Fluka; 10⁻³ mol/L), NaCl (4.5 g/L), and pH 6.7 ± 0.1 (Marichal et al., 1986 (link))]. The assay was then carried out as previously described by Pinto et al. (2013) (link). Briefly, C. albicans suspension was adjusted to a density of 1.0 ± 0.2 × 10⁶ CFU/mL and 990 μL of this suspension was distributed to test tubes. To each tube, 10 μL of sub-inhibitory concentrations of each essential oil and their major compounds were added and the cells were incubated without agitation for 3 h. Then, germ tube formation was registered under a light microscope (40×). Germ tubes were considered when the germinating protuberance was at least as long as the diameter of the blastopore. The conventional antifungal drug, fluconazole, was also tested and DMSO in a maximum concentration of 1% (v/v) was used as a control. The results are presented as the mean ± standard deviation (SD) of three independent experiments.

Cinnabarinic acid (CA, ≥98%, CAS: 606–59-7), Ehrlich reagent (CAS: 100–10-7), and N-acetylglucosamine (CAS: 7512-17-6) were purchased from Sigma-Aldrich (Shanghai, China). Sugarcane bagasse, wheat straw, rice straw, water hyacinth, and wood chip were purchased from a local market and dried to constant weight at 60°C before use. Other chemical reagents, unless otherwise specified, were all analytical grade.
Indicator strains, including Bacillus subtilis SYBC-hb1, Bacillus subtilis SYBC-hb5, Bacillus licheniformis SYBC-hb2, Bacillus pumilus SYBC-hb4, Bacillus thuringiensis SYBC-hb7, Bacillus amyloliquefaciens Y1-A3, Bacillus megaterium H021-A1, Bacillus cereus SYBC-hb8, Lysinibacillus sp. H021-S8, Staphylococcus kloosii H008-B4, Staphylococcus aureus, Escherichia coli J159, Serratia sp. L1, Serratia sp. L2, and Serratia sp. L3, were obtained from Biocatalysis and Transformation Biology lab at Jiangnan University (Wuxi, China).

P. falciparum strain NF54 was maintained in RPMI [7.4] supplemented with 10% human serum and 5% haematocrit using standard culturing technique^{87 (link)}. NF54/iGP2 strain was additionally supplemented with 2.5 mM D-(+)-glucosamine hydrochloride (Sigma #1514) for maintenance. For induction, parasites were synchronized with 5% sorbitol^{88 (link)} and glucosamine was omitted from the medium for 48 h. From day 4–8 gametocyte cultures were treated with 50 mM N-acetylglucosamine (Sigma #A6525) to eliminate ABS parasites^{89 (link)}. For WT-NF54 gametocyte induction the same procedure was followed except that instead of glucosamine induction, parasites were overgrown for 5 days with only medium exchanges every 48 h prior to N-acetylglucosamine treatment. At day 14 after induction, gametocytes were magnet-purified according to the procedure described previously^{90 (link)}. ABS parasites were not further enriched prior to processing. Infected red blood cells were freed from host material through 10 min incubation in 10× pellet volume of 0.05% (w/v) saponin in phosphate-buffered saline (PBS; pH 7.4) and subsequent centrifugation at 3000 × g for 5 min. The parasite pellet was washed twice with PBS and the dry pellet was flash frozen and stored at −80 °C.