G. Subramanian

Nitrogen stress induced changes in the marine cyanobacterium Oscillatoria willei BDU 130511

Exclusion of combined nitrogen (NaNO3) from the growth medium caused certain changes in metabolic processes leading to cessation in growth of the non-heterocystous, non nitrogen-fixing marine cyanobacterium Oscillatoria willei BDU 130511. But antioxidative enzymes, namely superoxide dismutase and peroxidase, helped the organism to survive the nitrogen stress. Prominent effects observed during nitrogen starvation/limitation were: (i) reduction of major and accessory photosynthetic pigments, (ii) impairment of photosynthesis due to loss of one major Rubisco isoenzyme, (iii) reduced synthesis of lipids and fatty acids, (iv) modifications of protein synthesis leading to the repression of three polypeptides and synthesis of two new polypeptides, (v) enhanced glutamine synthetase and reduced nitrate reductase activities, (vi) enhanced production of hydrogen peroxide and (vii) induced appearance of four new peroxidase isoenzymes. The observed metabolic changes were reversible, and the arrested growth under prolonged nitrogen deficiency could be fully restored upon subculturing in freshly prepared ASN III medium containing nitrogen (NaNO3). The present study demonstrates the capability of a non-nitrogen-fixer to withstand nitrogen stress making it an ecologically successful organism in the marine environment. The above pleiotropic effects of nitrogen deficiency also demonstrate that nitrogen plays a crucial role in growth and metabolism of marine cyanobacteria.

Cyanobacteria-dominated biofilms: a high quality food resource for intertidal grazers

Hong Kong rocky shores are dominated by cyanobacterial biofilms composed of a diversity of species. Thirteen common species, belonging to seven genera, were isolated in pure culture in MN+ and MN− media under defined growth conditions from a semi-exposed shore in Hong Kong. The nutritional values (i.e., protein, carbohydrate and calorific value) of these 13 species were determined. All species showed high nutritional quality in terms of protein, carbohydrate and calorific value, however, overall nutritional value varied between the species. Species of Spirulina and Phormidium were most nutritious (highest nutritional values) whereas species of Calothrix and Lyngbya were the least nutritious. Microphagous molluscan grazer density and diversity were relatively high at the study site, despite the seemingly low biomass (as assessed by chlorophyll a concentration) of the biofilm. It is suggested that the high nutritional quality of cyanobacteria, together with their fast turnover rates can support high levels of secondary production (biomass of grazers). The high nutritional quality of cyanobacteria on tropical, cyanobacteria-dominated, rocky shores is therefore of great importance in the benthic food web.

Ligninolytic and antioxidative enzymes of a marine cyanobacterium Oscillatoria willei BDU 130511 during Poly R-478 decolourization

Removal of combined nitrogen and addition of Poly R-478 to the growth medium enhanced oxidative stress, and altered the activities of ligninolytic enzymes of Oscillatoria willei BDU 130511. The activities of ligninolytic and antioxidative enzymes (LiP-like, LAC, PPO, SOD, POD, CAT, and APX) were increased upon nitrogen limitation and dye supplementation. The metabolic enzymes tested (GR, GPX, EST, and MDH) showed differential expressions under varied growth conditions. Up on nitrogen limitation, O. willei BDU 130511 showed enhanced ligninolytic activity as shown by alpha-keto-gamma-methylthiolbutyric acid (KTBA) oxidation and increased H(2)O(2) production. The organism decolourized 52% of Poly R-478 due to partial degradation and adsorption of dye particles from dye-added medium after 7 days of growth. This manuscript discusses the responses of ligninolytic and antioxidative enzymes of O. willei BDU 130511 during Poly R-478 decolourization/degradation, and the organisms potential in bioremediation.

Characterization of manganese superoxide dismutase from a marine cyanobacterium Leptolyngbya valderiana BDU20041

BackgroundCyanobacteria are recognized as the primordial organisms to grace the earth with molecular oxygen ~3.5 billion years ago as a result of their oxygenic photosynthesis. This laid a selection pressure for the evolution of antioxidative defense mechanisms to alleviate the toxic effect of active oxygen species (AOS) in cyanobacteria. Superoxide dismutases (SODs) are metalloenzymes that are the first arsenal in defense mechanism against oxidative stress followed by an array of antioxidative system. Unlike other living organisms, cyanobacteria possess multiple isoforms of SOD. Hence, an attempt was made to demonstrate the oxidative stress tolerance ability of marine cyanobacterium, Leptolyngbya valderiana BDU 20041 and to PCR amplify and sequence the SOD gene, the central enzyme for alleviating stress.ResultL. valderiana BDU 20041, a filamentous, non-heterocystous marine cyanobacterium showed tolerance to the tested dye (C.I. Acid Black 1) which is evident by increased in biomass (i.e.) chlorophyll a. The other noticeable change was the total ROS production by culture dosed with dye compared to the control cultures. This prolonged incubation showed sustenance, implying that cyanobacteria maintain their antioxidant levels. The third significant feature was a two-fold increase in SOD activity of dye treated L. valderiana BDU20041 suggesting the role of SOD in alleviating oxidative stress via Asada-Halliwell pathway. Hence, the organism was PCR amplified for SOD gene resulting in an amplicon of 550 bp. The sequence analysis illustrated the presence of first three residues involved in motif; active site residues at H4, 58 and D141 along with highly conserved Mn specific residues. The isolated gene shared 63.8% homology with MnSOD of bacteria confirmed it as Mn isoform. This is the hitherto report on SOD gene from marine cyanobacterium, L. valderiana BDU20041 of Indian subcontinent.ConclusionGeneration of Reactive Oxygen Species (ROS) coupled with induction of SOD by marine cyanobacterium, L. valderiana BDU20041 was responsible for alleviating stress caused by an azo dye, C. I. Acid Black 1. The partial SOD gene has been sequenced and based on the active site, motif and metal specific residues; it has been identified as Mn metalloform.

Oxygen-free hydrogen production by the marine cyanobacterium Phormidium valderianum BDU 20041

Abstract To circumvent the co-production of oxygen during hydrogen photoproduction, certain physico-chemical conditions were manipulated. Then, under white light (intensity 5.5 μmol photon m−2 s−1, 18 h dark/6 h light cycle); temperature 27°C; pH 7.5; salinity 25 g l−1, the marine, filamentous, non-heterocystous cyanobacterium Phormidium valderianum BDU 20041 was found to produce a maximum of 0.2 μmol hydrogen h−1 mg−1 dry wt in the gas phase without oxygen co-production; this was on a par with the maximum yield reports in the literature. To compensate for possible reduction in CO2 fixation under these conditions, when 10 m m of different carbohydrates were used, those other than galactose, lactose and trehalose were found to be favourable. For continuous production of hydrogen when amino acids were added as a nitrogen source, isoleucine was found to be the least inhibitory. The organism was found to be essentially a dark hydrogen producer.

Analysis and Elucidation of Phosphoenolpyruvate Carboxylase in Cyanobacteria

Phosphoenolpyruvate carboxylase (PEPC) a cytosolic enzyme of higher plants is also found in bacteria and cyanobacteria. Genetic and biochemical investigations have indicated that there are several isoforms of PEPC belonging to C3; C3/C4 and C4 groups but, the evolution of PEPC in cyanobacteria is not yet understood. The present study opens up an opportunity to understand the isoforms and functions of PEPC in cyanobacteria. The variations observed in PEPC among lower and higher orders of cyanobacteria, suggests convergent evolution of PEPC. There is a specific PEPC phosphorylation residue ‘serine’ at the N-terminus and PEPC determinant residue ‘serine’ at the C-terminal that facilitates high affinity for substrate binding. These residues were unique to higher orders of cyanobacteria, but, not in lower orders and other prokaryotes. The different PEPC forms of cyanobacteria were investigated for their kinetic properties with phosphoenolpyruvate as the substrate and the findings corroborated well with the in silico findings. In vitro enzymatic study of cyanobacteria belonging to three different orders demonstrated the role of aspartate as an allosteric effector, which inhibited PEPC by interacting with the highly conserved residues in the active site. The differences in mode of inhibition among the different order, thus, give a fair picture about the cyanobacterial PEPCs. The higher orders appear to possess the sequence coordinates and functionally conserved residues similar to isoforms of C4 type higher plants, whereas isoforms of PEPC of the lower orders did not resemble either that of C3 or C4 plants.