Biotechnological potential of Chlorella vulgaris for accumulation of Cu and Ni from single and binary metal solutions

This study provides information on the mechanism(s) of Cu and Ni ion biosorption by C. vulgaris, distinguishing adsorption from intracellular accumulation under various conditions. Surface adsorption was found to contribute maximally (>70%) to total Cu/Ni ion accumulation by the test alga (total accumulation efficiencies were 60 and 53 g metal ion mg protein^–1, respectively for Cu and Ni). Maximum intracellular uptake was reported at a pH range of 6.5–7.5, whereas adsorption reached its maximum at pH 3.5 for Cu, and pH 3.5 and 6.5 in the case of Ni. 35 °C was found to be the best temperature for maximum adsorption, whereas intracellular uptake was highest at 25 °C. Though exponentially grown C. vulgaris registered maximum metal ion uptake, adsorption maxima reached the highest values in the declining phase of the culture. Heat-killed and air-dried C. vulgaris accumulated Cu and Ni at about 80% of the values for viable samples, whereas formaldehyde-treated and immobilized biomasses depicted better accumulating potential than the control cells. Na, K, Mn and Zn caused competitive inhibition, whereas for Ca a mixed-type inhibition was evident. Thus, the present study suggests that the general concept that cations inhibit metal ion accumulation by competing with them for the same binding sites on the cell surface is not absolutely valid. As these results also demonstrate that a large amount of the bound metal (>70%) is in the adsorbed fraction, it is advantageous in the sense that it could be recovered by a suitable desorbing agent, especially in case of precious metals and the biomass could be exploited for repeated use in reactors.     

Process optimization for poly-beta-hydroxybutyrate production in a nitrogen fixing cyanobacterium, Nostoc muscorum using response surface methodology

A five-level-four-factor central composite rotary design was employed to find out the interactive effects of four variables, viz. concentrations of acetate, glucose and K2HPO4, and dark incubation period on poly-beta-hydroxybutyrate (PHB) production in a N2-fixing cyanobacterium, Nostoc muscorum. Acetate, glucose and dark incubation period exhibited positive impacts on PHB yield. Using response surface methodology (RSM), a second order polynomial equation was obtained by multiple regression analysis. A yield of 45.6% of dry cell weight (dcw) was achieved at reduced level of nutrients, i.e. 0.17% acetate, 0.16% glucose and 5 mg l(-1) K2HPO4 at a dark incubation period of 95 h as compared to 41.6% PHB yield in 0.4% acetate, 0.4% glucose and 40 mg l(-1) K2HPO4 at a dark incubation period of 168 h under single factor optimization strategy.     

Cloning and nucleotide sequence of a full-length cDNA encoding Ascaris suum phosphofructokinase

The nucleotide sequence of a full-length cDNA encoding phosphofructokinase (PFK) enzyme from the parasitic nematode Ascaris suum was determined. The entire sequence of 2,653 bases comprises a single open reading frame of 2,452 bases and a noncoding region of 201 bases after the stop codon. The mature protein contains 812 amino acids and has a molecular mass of 90,900 Da. The amino acid sequences of several peptides derived from the purified protein show excellent correspondence with the translated nucleotide sequence. Comparison of the amino acid sequence of the protein with those of 3 other worms as well as those of human, rabbit, and bacterial enzymes reveals highly conserved regions interrupted with stretches of lesser sequence similarity. Analyses of the subunit primary structure reveal, as in other eukaryotic PFKs, that the amino-terminal half is homologous to the carboxy-terminal half, supporting the hypothesis that the PFK gene evolved by duplication of the prokaryotic gene and that the allosteric sites arose by mutations at the catalytic site. The location of the phosphorylation site is unique and different compared with other PFKs and plays a key role in regulation of the enzyme activity. Structural motifs such as the putative substrate and effector binding domains and also the key amino acids involved therein are clearly identified by alignment of all the PFK protein sequences.     

Impact of aluminium,fluoride and fluoroaluminate complex on ATPase activity ofNostoc linckia andChlorella vulgaris

This study demonstrates a pH-dependent inhibition of Mg²⁺- and Ca²⁺- ATPase activities ofNostoc linckia andChlorella vulgaris exposed to AlCl₃, AlF₃, NaF and AlCl₃ + NaF together. AlF₃ and the combination of AlCl₃ + NaF were more inhibitory to both the enzymes as compared with AlCl₃ and NaF. Toxicity of the test compounds increased with increasing acidity. Interaction of AlCl₃ + NaF was additive onN. linckia andC. vulgaris, respectively, at pH 7.5 and 6.8, and synergistic at pH 6.0 and 4.5. In the presence of 60 and 100 m PO₄ ^3- an increased NaF concentration (in the AlCl₃ + NaF combination) was required to produce the same degree of inhibition in ATP synthesis and ATPase activity. Toxicity of fluoroaluminate was reduced in the presence of EDTA and citrate. Except for beryllium to some extent, combinations of cadmium, cobalt, iron, manganese, tin and zinc with fluoride were not as effective as aluminium in inhibiting the ATPase activity. The presence of a 100 kDa protein band in SDS-PAGE of both control as well as AlCl₃ + NaF-treated samples suggested that AlF₄ ^– inhibits the ATPase activity by acting as a functional barrier without affecting the structure of the enzyme.     

Carbon dioxide and poultry waste utilization for production of polyhydroxyalkanoate biopolymers by <Emphasis Type="Italic">Nostoc muscorum</Emphasis> Agardh: a sustainable approach

The new paradigm is to view wastes as resources for sustainable development. In this regard, the feasibility of poultry waste and CO₂ utilization for cultivation of a filamentous nitrogen-fixing cyanobacterium, Nostoc muscorum Agardh, was investigated for production polyhydroxyalkanoates, the biodegradable polymers. This cyanobacterium showed profound rise in biomass yield with up to 10 % CO₂ supply in airstream with an aeration rate of 0.1 vvm. Maximum biomass yield of 1.12 g L^?1 was recorded for 8 days incubation period, thus demonstrating a CO₂ biofixation rate of 0.263 g L^?1 day^?1 at 10 % (v/v) CO₂-enriched air. Poultry litter (PL) supplementation also had a positive impact on the biomass yield. The nutrient removal efficiency of N. muscorum was reflected in the significant reduction in nutrient load of PL over the experimental period. A maximum poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) [P(3HB-co-3HV)] copolymer yield of 774 mg L^?1 (65 % of dry cell wt.), the value almost 11-fold higher than the control, was recorded in 10 g L^?1 PL-supplemented cultures with 10 % CO₂ supply under the optimized condition, thus demonstrating that N. muscorum has good potential for CO₂ biomitigation and poultry waste remediation while simultaneously producing eco-friendly polymers.     

Response ofAnabaena doliolum to bimetallic combinations of Cu,Ni and Fe with special reference to sequential addition

This study reports physiological features of a N₂-fixing cyanobacteriumAnabaena doliolum in response to metal mixtures. Exposure of the cyanobacterium to Cu, Ni and Fe individually, as well as in combinations (Cu + Ni, Cu + Fe, Ni + Fe), showed marked differences in growth inhibition, nutrient uptake (NH₄ ⁺ and NO₃ ⁻), photosynthesis, ATP content, nitrate reductase, glutamine synthetase and urease activities. The response to metal combinations was also dependent upon the order in which the metals were added. The Cu-Ni combination resulted in synergistic interaction, in contrast to the antagonism of Cu-Fe and Ni-Fe. Pre-addition of Fe protected the cyanobacterium against Cu and Ni toxicity. Statistically significant (P < 0.005) inhibition of all the processes following metal supplementation was observed. This study suggests that carbon fixation is the most suitable variable for assessing heavy metal toxicity.