Polyhydroxybutyrate (PHB) accumulation by a mangrove isolated cyanobacteria Limnothrix planktonica using fruit waste.

Cyanobacterial polyhydroxybutyrate (PHB) is preferred over bacteria for low-cost production due to its photoautotrophic nature and lower carbon requirement. Considering its impact on the environment and circular economy, the valorization of fruit waste is the need of the hour. In the present study, fruit peels of banana, orange, pea, jackfruit, watermelon and waste flowers were tried as carbon sources for mangrove-isolated cyanobacteria Limnothrix planktonica to accumulate PHB. Alterations in the ASN-III culture medium and the introduction of untreated and pre-treated (acid/alkali-treated) peels as carbon sources are tried to enhance PHB. Banana peel showed the maximum PHB accumulation potential of 25.73 mg/L on the 12th day of incubation, followed by jackfruit (22.46 mg/L) and watermelon peels (20.72 mg/L); whereas, commercial carbon sources showed lower PHB accumulation up to 19.26 mg/L and 18.21 mg/L with fructose and glucose respectively. PHB accumulation was boosted to 5-fold higher (39.39 mg/L) in NP deficiency medium along with banana peel supplement, as compared to photoautotrophic conditions (8.49 mg/L) after the 9th day of incubation. Additionally, the PHB obtained by using the fruit wastes has a higher molecular weight than the PHB accumulated during photoautotrophic conditions. Optimization of parameters using fruit wastes and characterization of PHB would lead to its potential use.

Biofertilizers and nanofertilizers for sustainable agriculture: Phycoprospects and challenges.

Increased food demands and ceasing nutrient deposits have resulted in a great shortfall between the food supply and demand and would be worse in the years to come. Higher inputs of synthetic fertilizers on lands have resulted in environmental pollution, persistent changes in the soil ecology, and physicochemical conditions. This has greatly decreased the natural soil fertility thereby hindering agricultural productivity, human health, and hygiene. Bio-based resilient nutrient sources as wastewater-derived algae are promising as a complete nutrient for agriculture and have the potential to be used in soilless cultivations. Innovations in nano-fortification and nano-sizing of minerals and algae have the potential to facilitate nutrients bioavailability and efficacy for a multifold increase in productivity. In this context, various options on minerals nanofertilizer application in agricultural food production besides efficient biofertilizer have been investigated. Algal biofertilizer with the nanoscale application has huge prospects for further agriculture productivities and fosters suitable development.

Enhanced biomass production of Synechocystis sp. PCC 6803 by two associated bacteria Paenibacillus camelliae and Curtobacterium ammoniigenes.

Synechocystis sp. PCC 6803 grown continuously in a 5-L photo-bioreactor for 20 months was found to have associated consortia with heterotrophic microorganisms. Two strains of bacteria were isolated from the long-term cultures of cyanobacteria with the aim to test whether their presence affects cyanobacterial growth and metabolism. The two strains were phylogenetically identified as Paenibacillus camelliae and Curtobacterium ammoniigenes, respectively. Co-culturing the Synechocystis sp. with either of the isolates under photoautotrophic and photoheterotrophic conditions exerted a statistically significant growth enhancement effect on cyanobacteria. Under co-culture experiments, the addition of P. camelliae resulted in a four-fold higher biomass yield with a considerable decrease in the stationary period. The growth was more pronounced on the addition of acetate to the culture media. Growth-enhancing factors like indole acetic acid (IAA) and siderophores were detected in the co-culture conditions which proved to be the main driving force in boosting cyanobacterial growth. Thus, the cyanobacteria-bacteria consortia can be very useful for augmenting biomass production by circumventing the time factor which can be further exploited for various biotechnological applications.

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.

Studies on poly-beta-hydroxybutyrate synthase activity of Nostoc muscorum.

This study compares the PHB synthase activity of Nostoc muscorum, a N(2)-fixing cyanobacterium under control (grown in usual BG-11 medium), nitrogen (N) and phosphorus (P) deprivation and chemoheterotrophic conditions. Specific activity of PHB synthase did not depict significant variations in the latter three types of cultures, except for the control one, where a significantly lower activity was recorded. PHB synthase activity was detected only in the soluble fractions of both the control as well as cells incubated under chemoheterotrophic conditions. A K(m) of 80.2 microM DL-beta-hydroxybutyryl-CoA and V(max) of 197.5 nmol thiobenzoate (TNB) mg protein(-1)min(-1) were observed for the enzyme. PHB synthase remained insensitive to acetyl-CoA, ATP, NADP, NADPH supplementation under in vitro condition. Addition of acetyl phosphate was found to activate the enzyme and the level of activation was dependent on the concentration of acetyl phosphate supplementation. Inhibition of PHB synthase in 2,3-butanedione supplemented cultures and reactivation following acetyl phosphate addition proved the post-translational control of acetyl phosphate over PHB synthase.

Optimization of cultural and nutritional conditions for accumulation of poly-beta-hydroxybutyrate in Synechocystis sp. PCC 6803.

Poly-beta-hydroxybutyrate (PHB) accumulation in the unicellular cyanobacterium, Synechocystis sp. PCC 6803, was studied under various cultural and nutritional conditions. Under controlled condition, cells harvested at the stationary phase of growth depicted maximum accumulation of PHB, i.e., 4.5% (w/w of dry cells) as compared to lag (1.8%) or logarithmic (2.9%) phases of cultures. A temperature range of 28-32 degrees C and pH between 7.5 and 8.5 were preferred for PHB accumulation. Cells cultivated under regular light-dark cycles accumulated more PHB (4.5%) than those grown under continuous illumination (2.4%). Nitrogen and phosphorus starvation stimulated PHB accumulation up to the tune of 9.5 and 11% (w/w of dry cells), respectively. Synechocystis cells pre-grown in glucose (0.1%)-supplemented BG-11 medium when subjected to P-deficiency in presence of acetate (0.4%), PHB accumulation was boosted up to 29% (w/w of dry cells), the value almost 6-fold higher with respect to photoautotrophic condition. Fishpond discharges were found as suitable media for PHB accumulation in the test cyanobacterium.

Poly-beta-hydroxybutyrate accumulation in Nostoc muscorum and Spirulina platensis under phosphate limitation.

Nostoc muscorum and Spirulina platensis were grown under phosphate deficiency in order to investigate the role of internal phosphate pool and activity of alkaline phosphatase on poly-beta-hydroxybutyrate (PHB) accumulation. PHB accumulation in N. muscorum increased to 22.7% of dry weight (dw) after 4 day of phosphate deficiency, while the internal phosphate pool reduced to the level of 0.02 microM mg dw(-1) at a maximum APase activity of 2.57nM PNP mg dw(-1) h(-1). In contrary, S. platensis depicted maxima of 1.39nM PNP mg dw(-1) h(-1) on day 30 of incubation, which was about 2 fold lower than the observed value of N. muscorum. PHB content in S. platensis remained low even after prolonged phosphate starvation, and a rise only up to 3.5% of dw was recorded on day 60 of phosphate deficiency. Supplementation of NADPH exogenously to S. platensis cultures grown under phosphate deficiency favoured PHB accumulation in 10, 20 and 30 days old cultures, but not in the cultures grown under phosphate deficiency for 60 days. The possible role of phosphate limitation on PHB accumulation is discussed.