Microbial growth on peptones from fish industrial wastes

Industrial fish peptone was an excellent substrate for biomass production in solid and submerged fermentations. The maximal growth rates of several microorganisms were two to three times higher than those grown on beef (bacto)peptones and the final biomass concentrations were almost twice as great as those grown on beef peptones. Fish peptones did not increase the production of secondary metabolites relative to those produced on beef peptones in non-optimized media. Fish peptone has promising potential as a substrate for biomass production.     

High-throughput screening for potent and selective inhibitors of Plasmodium falciparum dihydroorotate dehydrogenase

Plasmodium falciparum is the causative agent of the most serious and fatal malarial infections, and it has developed resistance to commonly employed chemotherapeutics. The de novo pyrimidine biosynthesis enzymes offer potential as targets for drug design, because, unlike the host, the parasite does not have pyrimidine salvage pathways. Dihydroorotate dehydrogenase (DHODH) is a flavin-dependent mitochondrial enzyme that catalyzes the fourth reaction in this essential pathway. Coenzyme Q (CoQ) is utilized as the oxidant. Potent and species-selective inhibitors of malarial DHODH were identified by high-throughput screening of a chemical library, which contained 220,000 drug-like molecules. These novel inhibitors represent a diverse range of chemical scaffolds, including a series of halogenated phenyl benzamide/naphthamides and urea-based compounds containing napthyl or quinolinyl substituents. Inhibitors in these classes with IC(50) values below 600 nm were purified by high pressure liquid chromatography, characterized by mass spectroscopy, and subjected to kinetic analysis against the parasite and human enzymes. The most active compound is a competitive inhibitor of CoQ with an IC(50) against malarial DHODH of 16 nm, and it is 12,500-fold less active against the human enzyme. Site-directed mutagenesis of residues in the CoQ-binding site significantly reduced inhibitor potency. The structural basis for the species selective enzyme inhibition is explained by the variable amino acid sequence in this binding site, making DHODH a particularly strong candidate for the development of new anti-malarial compounds.     

Identification of mammalian aspartate-4-decarboxylase

Several animal tissues were examined for aspartate-4-decarboxylase (EC 4.1.1.12) activity. Highest activity was seen in murine livers, in rodent livers, and in rodent kidneys. The rat liver enzyme was membrane associated and could be solubilized and partially purified with the aid of detergents. The purification studies, and studies on the stoichiometry and kinetics of the reaction, showed that aspartate is directly converted to alanine. Such a metabolic reaction had not been reported before in animals. The rat liver enzyme differed significantly from the microbial aspartate-4-decarboxylases. Among other things, the rat liver beta-decarboxylase could be purified away from a cysteine sulfinate desulfinase activity. Also, unlike the bacterial enzymes, the mammalian beta-decarboxylase could not be inactivated by preincubation with aspartate or cysteine sulfinate. These later observations strongly suggest that the mammalian aspartate-4-decarboxylase does not have an inherent transaminase activity. Like many decarboxylases, rat liver aspartate-4-decarboxylase could be inhibited by reagents which react with carbonyl groups; however, the enzyme showed no dependence on pyridoxal 5'-phosphate.     

Effect of various additives on enzymatic hydrolysis of castor oil

Hydrolysis of castor oil using lipase enzyme is carried out in a batch reactor at room temperature (35–40 °C). In order to reduce the cost of enzyme catalyzed reaction, water in oil emulsion and a 3:1 ratio of oil to water is selected. The concentration of enzyme in the reaction mixture is optimized. The effect of various additives like solvent and salt which can enhance the rate of reaction is studied. It is found that the glycerol has no effect on the hydrolysis of oil. The reusability of the lipase enzyme has also been tested. The yield of enzymatic hydrolysis of castor oil is compared with those of coconut oil and olive oil.     

Deformability limits of Plasmodium falciparum-infected red blood cells

Splenic filtration of infected red blood cells (RBCs) may contribute to innate immunity and variable outcomes of malaria infections. We show that filterability of individual RBCs is well predicted by the minimum cylindrical diameter (MCD) which is calculated from a RBC's surface area and volume. The MCD describes the smallest diameter tube or smallest pore that a cell may fit through without increasing its surface area. A microfluidic device was developed to measure the MCD from thousands of individual infected RBCs (IRBCs) and uninfected RBCs (URBCs). Average MCD changes during the blood-stage cycle of Plasmodium falciparum were tracked for the cytoadherent strain ITG and the knobless strain Dd2. The MCD values for IRBCs and URBCs raise several new intriguing insights into how the spleen may remove IRBCs: some early-stage ring-IRBCs, and not just late-stage schizont-IRBCs, may be highly susceptible to filtration. In addition, knobby parasites may limit surface area expansions and thus confer high MCDs on IRBCs. Finally, URBCs, in culture with IRBCs, show higher surface area loss which makes them more susceptible to filtration than naive URBCs. These findings raise important basic questions about the variable pathology of malaria infections and metabolic process that affect volume and surface area of IRBCs.     

Poly-extremotolerant bacterium isolated from reverse osmosis reject: an implication toward waste water management

We demonstrate the tolerance of bacterial strain SM2014 to various unsustainable conditions and suggest its implication in waste water management. Its sustainability to reverse osmosis pressure (2.1 MPa) during desalination, and survival percentage of 73 % under hyperbaric conditions (pressure tension of 3.1 MPa under absolute oxygen atmosphere) confirmed its pressure tolerance. The growth of this strain at pH 9 or 10 and at 60 °C alone or in combination revealed its unique physiology as poly-extremotolerant strain. As an adaptive mechanism, the ratio of saturated to unsaturated fatty acids changed with growth conditions. Under poly-extreme condition long chain saturated fatty acid (C_18:0, C_16:0, C_14:0, C_12:0) predominated at the expense of unsaturated fatty acids. The nucleotide BLAST of 16S rRNA gene sequence of strain SM2014 with the NCBI gene bank sequences showed its close identity to Bacillus licheniformis with a similarity match of 94 %. The secretion of industrially valuable enzymes proteinase, lipase and amylase under such harsh conditions further signified potential of this strain as a source of extremozymes. Its unique characteristics underscore its relevance in waste water management.