Screening of Anti-inflammatory Active Sites of Water Extract of Iris halophila Root and Analysis of Its Chemical Constituents / 中国实验方剂学杂志

Objective:To investigate the anti-inflammatory effects of water extract of the<italic> Iris halophila</italic> root on lipopolysaccharide（LPS） stimulated RAW264.7 cells and analyze its chemical constituents. Method:The supernatant of YWG prepared by water extraction and alcohol precipitation was separated by AB-8 macroporous adsorption resin column chromatography to obtain ethanol eluates with different concentrations （YWG，YWG-0%，YWG-20%，YWG-40%，and YWG-60%）. Cell counting kit-8（CCK-8） assay was used to determine the effects of YWG-0%，YWG-20%，YWG-40%，and YWG-60% on the viability of RAW264.7 cells. Griess assay was employed to detect the nitric oxide （NO） level in LPS-stimulated RAW264.7 cells. The release of tumor necrosis factor（TNF）-<italic>α</italic>，interleukin（IL）-6，IL-10，and IL-1<italic>β</italic> was detected by enzyme-linked immunosorbent assay（ELISA）. YWG and the elution site with the most robust anti-inflammatory activity were identified and compared by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-Q-TOF-MS/MS）. Result:Ethanol eluates with different concentrations inhibited the release of NO，TNF-α，IL-1<italic>β</italic>， and IL-6 in the supernatant of LPS induced RAW264.7 cells （<italic>P<</italic>0.05），and promoted the release of IL-10 （<italic>P<</italic>0.05）. YWG-60% displayed a highly significant effect （<italic>P</italic><0.01）. A total of 127 constituents were detected from the comparison of YWG and YWG-60% by UHPLC-Q-TOF-MS/MS in the positive and negative ion modes，including 61 flavonoids. YWG-60% contained 25 flavonoids with elevated content as compared with YWG. Conclusion:YWG-60% showed potent anti-inflammatory effect，and the effective anti-inflammatory constituents were presumedly flavonoids. The findings of this study are expected to provide a scientific theoretical basis for the basic research on the medicinal effect of the water extract of YWG.

Isolation and Characterization of Halophilic Bacterial Strains from Saline Waters of Khewra Salt Mines on the Basis of 16S rRNA Gene Sequence.

Halophiles are salt loving microbes optimally growing at high concentrations of salt. Khewra salt mines of Pakistan provide extreme saline conditions where enormous halophilic microbial biota thrives. The present study aimed at isolation and molecular identification of bacterial strains from saline waters of Khewra salt mines. Using halophilic media, nine halophilic bacterial strains from saline water bodies were cultured and studied under optimized growth conditions (NaCl, pH and temperature). Bacterial growth at different NaCl concentrations was measured at 600nm wavelength, showing optimal growth at 1.5M NaCl. 769bp size 16S rRNA gene was amplified for molecular identification of bacterial strains. The amplified genes of the strains FA2.2 and FA3.3 were sequenced and their homology with other bacterial strains was analyzed. The results showed FA2.2 shared maximum homology with Bacillus anthracis strain while FA3.3 showed close resemblance with Staphylococcus saprophyticus subsp. bovis. Isolated halophilic bacterial strains possess potential for various biotechnological applications. They could be manipulated for synthesizing transgenic crops tolerating high salinity boosting the agricultural yield. Moreover extremozymes of these bacteria holds great industrial importance.

Biodegradation of petroleum hydrocarbons in hypersaline environments

Literature on hydrocarbon degradation in extreme hypersaline media presents studies that point to a negative effect of salinity increase on hydrocarbonoclastic activity, while several others report an opposite tendency. Based on information available in the literature, we present a discussion on the reasons that justify these contrary results. Despite the fact that microbial ability to metabolize hydrocarbons is found in extreme hypersaline media, indeed some factors are critical for the occurrence of hydrocarbon degradation in such environments. How these factors affect hydrocarbon degradation and their implications for the assessment of hydrocarbon biodegradation in hypersaline environments are presented in this review.

Production of Halophilic α-Amylase by Immobilized Cells of Moderately Halophilic Bacillus sp. Strain TSCVKK.

Aims: To investigate the effect of cell immobilization on amylase production by the moderately halophilic bacterium, Bacillus sp. strain TSCVKK and to compare the properties of the amylase produced under immobilized conditions with the enzyme produced by the free cells. Study Design: Cell immobilization. Place and Duration of Study: Department of Chemistry, Biochemistry Lab, Indian Institute of Technology (IIT Madras), Chennai, Tamil Nadu, between Jan 2009 and March 2009. Methodology: Bacillus sp. strain TSCVKK was immobilized in alginate, agar, polyacrylamide and gelatin. Production of amylase was determined using 3, 5- dinitrosalicylic acid (DNS). Effect of NaCl, pH, temperature on the activity of amylase was determined and compared with the amylase produced by the free cells. Results: Maximum production of 832 mU/ml was achieved with an initial cell load of 1.2% (w/v; wet weight) of 24 h grown cells immobilized in 2% agar of 4 mm3 block size using GSL-2 medium containing 10% NaCl and 1.5% dextrin at pH 8.0 at 30ºC after 36 h of growth. Amylase production was lower when the cells were immobilized in alginate (211 mU/ml) or with the free cells of same biomass concentration as used for immobilization (333 mU/ml). Amylase was not produced when gelatin or polyacrylamide was used as the immobilization matrix. The immobilized cells in 2% agar could be used up to 5 cycles without much reduction in amylase production. Amylase produced through cell immobilization retained all the properties that were shown by amylase produced under submerged fermentation. Conclusion: Agar was the suitable matrix to immobilize Bacillus sp. strain TSCVKK for amylase production. Amylase produced under immobilization conditions retained its temperature, salt and pH requirements. Immobilized cells were used for 5 cycles without much decrease in production.