Synthesis of Feruloyl Ester Using Bacillus subtilis AKL 13 Lipase Immobilized on Celite® 545

U ovom su istraživanju sintetizirani lipofilni antioksidansi, i to gliceril ferulat i feruloil gliceril linoleat, pomoću lipaze izolirane iz bakterijskog soja Bacillus subtilis AKL 13. Ekstracelularna lipaza dobivena je kultivacijom soja u modificiranoj minimalnoj podlozi i taloženjem pomoću 80 %-tne amonijeve soli. Koncentrirani enzim specifične aktivnosti od (4647±66) U/mg imobiliziran je na dijatomejskoj zemlji Celite® 545 i unakrsno povezan pomoću glutaraldehida. Pripremljeni enzimski katalizator upotrijebljen je za esterifikaciju ferulinske i zasebno linoleinske kiseline pomoću glicerola u heksanu/butanu pri 50 °C uz miješanje (3.144×g). Maksimalna konverzija feruloil gliceril linoleata od 94 % postignuta je tijekom 48 h, dok je ona gliceril ferulata bila 35 %. Produkti reakcije okarakterizirani su pomoću RP-HPLC, FT-IR, 1H NMR, 13C NMR metoda i fluorescencijskog spektrofotometra. Kinetički parametri reakcije esterifikacije određeni su prema ping-pong bi-bi modelu. Vrijednosti Km i vmax za gliceril ferulat iznosile su 69.37 mmol i 0.387 mmol/(min∙g), a za feruloil gliceril linoleat 3.46 mmol i 1.02 mmol/(min∙g). Kinetički parametri simulirani su u programskom jeziku MATLAB, a eksperimentalni podaci uspješno su se slagali s teorijskim vrijednostima. Nadalje, sposobnost uklanjanja difenil-1-pikril-hidrazil radikala bila je veća u palminom ulju pomiješanom s esterima ferulinske kiseline nego u čistom palminom ulju i sličnija vrijednosti α-tokoferola.The lipophilic antioxidants, glyceryl ferulate and feruloyl glyceryl linoleate, were synthesized using lipase from Bacillus subtilis AKL 13. The extracellular lipase was produced by cultivation of the strain in modified minimal medium and the enzyme was recovered by fractionation at 80 % ammonium salt saturation. The concentrated enzyme with the specific activity of (4647±66) U/mg was immobilized on Celite® 545 and crosslinked using glutaraldehyde. The prepared enzyme catalyst was used for esterification of ferulic and linoleic acids with glycerol separately in hexane butane solvent system at 50 °C and 3.144×g agitation. The maximum ester conversion of 94 % of feruloyl glyceryl linoleate was achieved at 48 h, whereas only 35 % of glyceryl ferulate was synthesized. The reaction products were characterized using RP-HPLC, FTIR, 1H NMR, 13C NMR and fluorescence spectrophotometry. The kinetic parameters of esterification reaction were determined according to ping-pong bi-bi model. The Km and υmax were found to be 69.37 and 3.46 mmol, and 0.387 and 1.02 mmol/(min·g) for glyceryl ferulate and feruloyl glyceryl linoleate, respectively. The kinetic parameters were simulated in MATLAB and the experimental data were in good agreement. Furthermore, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of the blend of feruloyl ester and palm oil was higher than of the plain palm oil and was closer to α-tocopherol

Anticoagulant property of sulphated polysaccharides extracted from marine brown algae collected from Mandapam Island, India

The marine brown algae: Sargassum tenerrimum, Sargassum wightii, Turbinaria conoides, Turbinaria ornata and Padina tetrastromatica were collected from Mandapam Island, India. The crude sulphated polysaccharides (SPS) were extracted using hot water and examined for anticoagulation activity. The sugar, sulphate and protein in crude SPS were analyzed. The presence of sulphated polysaccharide was confirmed by agarose gel electrophoresis and further characterized by Fourier transform infrared spectroscopy (FTIR). The FTIR analysis of crude SPS showed characteristic band of polysaccharides at 900, 1740 cm-1 and ester sulphate at 1250 to 1260 cm-1. Moreover, the absorbance band at 820 cm-1 for S. tenerrimum, S. wightii, T. conoides and T. ornata denotes sulphation at equatorial position, but in the case of P. tetrastromatica, sulphation at axial position is denoted by absorbance band at 850 cm-1. The heparin like activity of crude SPS was determined by metachromatic assay. The anticoagulant activity of crude SPS was evaluated by activated partial thromboplastin time (APTT) and prothrombin time (PT) assays. The metachromatic, APTT and heparinoid activities of crude SPS from S. tenerrimum, S. wightii, T. conoides, T. ornata and P. tetrastromatica were in the range of 0.045 to 0.0347, 134 to 89 s and 25.47 to 14.5 USP units/mg, respectively. The prolongation of prothrombin time by crude SPS was not found.Keywords: Brown algae, sulphated polysaccharides, Fourier transform infrared spectrometer (FTIR), agarose gel, metachromatic activity, activated partial thromboplastin time (APTT), heparinAfrican Journal of Biotechnology Vol. 12(16), pp. 1937-194

Synthesis of Feruloyl Ester Using Bacillus subtilis AKL 13 Lipase Immobilized on Celite® 545

The lipophilic antioxidants, glyceryl ferulate and feruloyl glyceryl linoleate, were synthesized using lipase from Bacillus subtilis AKL 13. The extracellular lipase was produced by cultivation of the strain in modified minimal medium and the enzyme was recovered by fractionation at 80 % ammonium salt saturation. The concentrated enzyme with the specific activity of (4647±66) U/mg was immobilized on Celite® 545 and crosslinked using glutaraldehyde. The prepared enzyme catalyst was used for esterification of ferulic and linoleic acids with glycerol separately in hexane butane solvent system at 50 °C and 3.144×g agitation. The maximum ester conversion of 94 % of feruloyl glyceryl linoleate was achieved at 48 h, whereas only 35 % of glyceryl ferulate was synthesized. The reaction products were characterized using RP-HPLC, FTIR, 1H NMR, 13C NMR and fluorescence spectrophotometry. The kinetic parameters of esterification reaction were determined according to ping-pong bi-bi model. The Km and υmax were found to be 69.37 and 3.46 mmol, and 0.387 and 1.02 mmol/(min·g) for glyceryl ferulate and feruloyl glyceryl linoleate, respectively. The kinetic parameters were simulated in MATLAB and the experimental data were in good agreement. Furthermore, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of the blend of feruloyl ester and palm oil was higher than of the plain palm oil and was closer to α-tocopherol

Influence of Biosurfactant in Biosynthesis of Silver Nanoparticles by Pseudomonas Aeruginosa AMB AS7

ABSTRACT-The present study demonstrates green process for the synthesis of Silver nanoparticles (AgNPs) by bacteria Pseudomonas aeruginosa AMB AS7 and the role of biosurfactant in enhancing the stability of AgNPs. An aqueous solution of silver nitrate was treated with both cell free broth and biomass of P. aeruginosa AMB AS7 which resulted in the synthesis of AgNPs. The nanometallic dispersions were characterized by surface plasmon spectrum measuring at a range of 280 to 700nm. The particle distribution analysis and Scanning Electron Microscopic (SEM) results showed the formation of AgNPs in the range of 30 -80 nm. Further analysis carried out by Fourier Transform Infrared Spectroscopy (FTIR) showed the presence of functional groups from proteins and biosurfactant. The stability of AgNPs synthesised by both chemical and biological methods was analysed. Biological method of AgNPs synthesis showed remarkable stability. They were stable for 24 days whereas AgNPs synthesised by chemical method were stable for only 3 days. Also the influence of biosurfactant and chemical surfactant on the stability of AgNPs synthesized by chemical method was analyzed. The biosurfactant showed superior property (19 days) over the chemical surfactant (7 days) in terms of increasing the stability of AgNPs. The use of bacterium for nanoparticles synthesis and the influence of biosurfactant on the stability of AgNPs offer the benefits of eco-friendliness and amenability for large-scale productio

Protective effect of crude sulphated polysaccharide from <em>Turbinaria ornata</em> on isoniazid rifampicin induced hepatotoxicity and oxidative stress in the liver, kidney and brain of adult Swiss albino rats

237-244The bio activation of drugs to chemically active compounds, induces liver injury resulting in various degenerative diseases, which could be overcome by the administration of sulphated polysaccharides. The present study addresses, the hepatoprotective potential of crude sulphated polysaccharides (CSP) from Turbinaria ornata using a Swiss albino rat model where liver damage was induced with Rifampicin (RMP) and Isoniazid (INH). The RMP-INH (75 mg/kg body weight) and CSP (10, 25, 50 and 200 mg/kg body weight) were orally administered to Swiss albino rat for 14 days. Silymarin (100 mg/kg body weight) administered group served as a positive control. At the end of the experiment serum was collected and the rats were dissected and the organs like liver, kidney and brain were collected. Administration of hepatotoxin causes increase in serum SGPT, SGOT, lipid peroxidation levels and a decrease in reduced glutathione, GST levels. Co-administration of CSP shows hepatoprotective activity on hepatotoxin induced damage. The hepatoprotective activity was also supported by histopathological studies of liver sections. In addition, our study revealed that the effect of toxicity induced in liver also caused damage to kidney and brain tissues resulting in decrease of total antioxidant status, reduced glutathione, increased levels of nitrate and macromolecular damage like lipid peroxidation. Co-administration of CSP significantly prevents the damage of liver, kidney and brain caused by hepatotoxin. The CSP (25 mg/kg body weight) from T. ornata could be a better hepatoprotective agent than standard drug Silymarin