Effect of factors on pepsin and trypsin fibrinolytic activity and deactivation of fibrinolytic activity / 中草药

Objective: To investigate the method for study on the effect of factors on pepsin and trypsin fibrinolytic activity and deactivation of fibrinolytic activity and to eliminate the interference of pepsin and trypsin on the detection of crude protein fibrinolytic activity of Armadillidium vulgare (porcellio plasmin) in order to obtain the proteins or peptides which have the smaller molecular weight but higher titer during the pepsin and trypsin degradation. Methods: To study the effect of pepsin and trypsin deactivation on pH value, temperature, metal ions, enzyme inhibitor, surfactant, and responsing fibrinolytic by fiber fibrin plate assay. The better enzyme deactivation process was obtained and used for studying the effect on the fibrinolytic activity of urokinase, lumbrokinase, and porcellio plasmin. Results: All the pH value, temperature, metal ions, enzyme inhibitor, and surfactant have had an impact on pepsin and trypsin fibrinolytic activity. Among them the optimum deactivation of pepsin was pH 6.0-8.0, while the optimum deactivation of trypsin was mixed preparation with TLCK at the concentration of 25 mg/mL and EDTA at the concentration of 1 mmol/L. Conclusion: This study has obtained the better enzyme deactivation process which could be used for the detection of fibrinolytic activity of pepsin and trypsin degradation product by fiber fibrin plate assay, the operation is simple, and the repeatability and stability are good.

Bioprospecting of Cellulase Producing Extremophilic Bacterial Isolates from India.

Aim: To isolate and identify the potential extremophilic cellulase producing strain viz., psychrophiles, halophiles, thermophiles and to compare the Cellulase activity from samples collected from different geographical regions of India. Place and Duration of Study: Bharathiar University, Department of Biotechnology, Molecular Microbiology Lab, Coimbatore, Tamilnadu, India, between January to April 2011. Methodology: Cellulase-producing extremophilic bacteria viz., psychrophiles, halophiles, and thermophiles have been isolated from soil samples. According to morphology and pigmentation, 138 distinct bacteria were isolated and screened for cellulase activity by Gram’s iodine–carboxymethylcellulose plate (CMC) assay. On the basis of the cellulase activity, six potent cellulase-producing isolates from each cluster viz., P14, P36, H6, H13, T2 and T3 were selected for 16S rRNA gene based identification. The strains were optimized for maximum cellulase activity at various temperature and pH range. Results: The phylogenetic relationship revealed that P14 and P36 psychrophilic isolates possessed maximum identity with Bacillus simplex (100%) and Arthrobactercitreus (99%), with a cellulase activity of 14.10± 1.73 and 18.27± 0.71 UmL-1 respectively. Likewise, among halophiles, H6 and H13 were identified as Bacillus subtilis and Bacillus endophyticus (99%), with a cellulase activity of 14.87 ± 0.55 and 16.83 ± 0.44 U mL-1, correspondingly. In thermophiles, T2 and T3 showed close proximity with Bacillus amyloliquefaciens and Bacillus megaterium (99%), with a cellulase activity of 21.53 ± 1.30 and 19.93 ± 0.38 U mL-1 respectively. Conclusion: In the present study, the thermophilic isolates showed promising Cellulase activity compared to psychrophiles and halophiles.

A simple and rapid plate assay for the screening of Indole-3-Acetic Acid (IAA) producing microorganisms.

The indole-3-acetic acid (IAA) producing plant growth promoting rhizobacteria (PGPR) are routinely screened in the laboratory by colorimetric method in culture supernatant which is time consuming and expensive. We developed a novel plate assay for the detection of IAA in the bacterial strains. This method is rapid, cheap and accurate for IAA production in microorganisms.

Screening and characterization of protease from bacillus sp.

Protease production by alkalophilic Bacillus sp were isolated from different regions of Bangalore and used to screen for the protease production by using casein and skim milk agar plate assay. The agro wastes are also used to screen and produce protease. The agro wastes are rice bran, paddy straw and pigeon pea waste, among these substrate rice bran showed maximum synthesis of protease. At 48 hr it showed maximum 0.13 IU protease production in rice bran medium was used for partial purification studies. Fermented medium were used for salt precipitation, dialysis. The dayalyste were used for studies on effect of Ph, temperature, inhibitors and metal ions. The optimum pH 10, temperature 650C were optimum and the inhibitors and metal ion studies, metalions like Mn2+ and Ca2+ were found to be potent enhancers. The complete loss of enzyme activity was found in presence of PMSF.