Application of ANN-MOGA for nutrient sequestration for wastewater remediation and production of polyunsaturated fatty acid (PUFA) by Chlorella sorokiniana MSP1.

Chlorella bears excellent potential in removing nutrients from industrial wastewater and lipid production enriched with polyunsaturated fatty acids. However, due to the changing nutrient dynamics of wastewater, growth and metabolic activity of Chlorella are affected. In order to sustain microalgal growth in wastewater with concomitant production of PUFA rich lipids, RSM (Response Surface Methodology) followed by heuristic hybrid computation model ANN-MOGA (Artificial Neural Network- Multi-Objective Genetic Algorithm) were implemented. Preliminary experiments conducted taking one factor at a time and design matrix of RSM with process variables viz. Sodium chloride (1 mM-40 mM), Magnesium sulphate (100 mg-800 mg) and incubation time (4th day to 20th day) were validated by ANN-MOGA. The study reported improved biomass and lipid yield by 54.25% and 12.76%, along with total nitrogen and phosphorus removal by 21.92% and 18.72% respectively using ANN-MOGA. It was evident from FAME results that there was a significantly improved concentration of linoleic acid (19.1%) and γ-linolenic acid (21.1%). Improved PUFA content makes it a potential feedstock with application in cosmeceutical, pharmaceutical and nutraceutical industry. The study further proves that C. sorokiniana MSP1 mediated industrial wastewater treatment with PUFA production is an effective way in providing environmental benefits along with value addition. Moreover, ANN-MOGA is a relevant tool that could control microalgal growth in wastewater.

Recent advances in enhancing the production of long chain omega-3 fatty acids in microalgae.

Omega-3 fatty acids have gained attention due to numerous health benefits. Eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) are long chain omega-3 fatty acids produced from precursor ALA (α-linolenic acid) in humans but their rate of biosynthesis is low, therefore, these must be present in diet or should be taken as supplements. The commercial sources of omega-3 fatty acids are limited to vegetable oils and marine sources. The rising concern about vegan source, fish aquaculture conservation and heavy metal contamination in fish has led to the search for their alternative source. Microalgae have gained importance due to the production of high-value EPA and DHA and can thus serve as a sustainable and promising source of long chain omega-3 fatty acids. Although the bottleneck lies in the optimization for enhanced production that involves strategies viz. strain selection, optimization of cultivation conditions, media, metabolic and genetic engineering approaches; while co-cultivation, use of nanoparticles and strategic blending have emerged as innovative approaches that have made microalgae as potential candidates for EPA and DHA production. This review highlights the possible strategies for the enhancement of EPA and DHA production in microalgae. This will pave the way for their large-scale production for human health benefits.

Exploring the structural aspects and therapeutic perspectives of cyanobacterial phycobiliproteins.

Phycobiliproteins (PBPs) of cyanobacteria and algae possess unique light harvesting capacity which expand the photosynthetically active region (PAR) and allow them to thrive in extreme niches where higher plants cannot. PBPs of cyanobacteria/algae vary in abundance, types, amino acid composition and in structure as a function of species and the habitat that they grow in. In the present review, the key aspects of structure, stability, and spectral properties of PBPs, and their correlation with ecological niche of cyanobacteria are discussed. Besides their role in light-harvesting, PBPs possess antioxidant, anti-aging, neuroprotective, hepatoprotective and anti-inflammatory properties, which can be used in therapeutics. Recent developments in therapeutic applications of PBPs are reviewed with special focus on 'route of PBPs administration' and 'therapeutic potential of PBP-derived peptide and chromophores'.

Nutritional, Functional, Textural and Sensory Evaluation of Spirulina Enriched Green Pasta: A Potential Dietary and Health Supplement.

In house cultivated Spirulina powder was incorporated at 2 to 15% concentrations to enrich pasta prepared from semolina. Spirulina incorporation led to development of green color pasta with nutritional and functional fortification resulting in increase in its protein, total phenols, flavonoids, iron and calcium content by up to 77.47%, 76.62%, 162.88%, 296.99% and 57.27%, respectively, without causing detrimental changes to the textural and sensory attributes. FAME analysis revealed 2 to 2.5 times enhanced levels of γ-linolenic acid and docosahexaenoic acid in enriched pasta. Significant improvement in phenolics, flavonoids and antioxidant activity were also observed in comparison to control pasta. Analysis of theoretical and realized composition confirmed retention of nutrients post cooking revealing no significant loss in proteins and other nutrients. Principal components analysis demonstrated significant contribution of Spirulina to nutritional and functional attributes especially at higher concentrations. Pasta enriched with 12.5% Spirulina was rated as "liked very much" and the purchase intention was also high. Spirulina enrichment at concentrations above 10% (12.5%) with appreciable increase in nutritional and functional attributes without affecting textural or cooking quality and acceptable sensory evaluation can be a preferred alternative to augment health and prevent sickness. Since green color symbolizes freshness, hope, renewal and physical health, the consumption of Spirulina incorporated green pasta may be a potential option to enhance the livelihood and nutritional security of rural poor and a good alternative for hidden hunger alleviation programs for mass nutrition especially for infants and children in an effective manner.

Biosynthesis and biotechnological interventions for commercial production of microalgal pigments: A review.

Microalgae are photosynthetic eukaryotes that serve as microbial cell factories for the production of useful biochemicals, including pigments. These pigments are eco-friendly alternatives to synthetic dyes and reduce environmental and health risks. They also exhibit excellent anti-oxidative properties, making them a useful commodity in the nutrition and pharmaceutical industries. Light-harvesting pigments such as chlorophylls and phycobilins, and photoprotective carotenoids are some of the most common microalgal pigments. The increasing demand for these pigments in industrial applications has prompted a need to improve their metabolic yield in microalgal cells. So far, expensive cultivation methods and sensitivity to microbial contamination remain the main obstacles to the large-scale production of these pigments. This review highlights current issues and future prospects related to the production of microalgal pigments. The review also emphasizes the use of engineering approaches such as genetic engineering, and optimization of media components and physical parameters to increase their commercial-scale production.

Effects of nanofertilizers on soil and plant-associated microbial communities: Emerging trends and perspectives.

Modern agricultural practices are relying excessively upon the use of synthetic fertilizers to supply essential nutrients to promote crop productivity. Though useful in the short term, their prolonged and persistent applications are harmful to soil fertility and nutrient dynamics of the rhizospheric microbiome. The application of nanotechnology in form of nanofertilizer provides an innovative, efficient, and eco-friendly alternative to synthetic fertilizers. The nanofertilizers allow a slow and sustained release of nutrients that not only supports plant growth but also conserve the diversity of the beneficial microbiome. Such attributes may help the phytomicrobiome to efficiently mitigate both biotic and abiotic stress conditions. Unfortunately, despite, exceptional efficiency and ease of applications, certain limitations are also associated with the nanofertilizers such as their complicated production process, tenuous transport and dosage-sensitive efficiency. These bottlenecks are causing a delay in the large-scale applications of nanofertilizers in agriculture. This review aims to highlight the current trends and perspectives on the use of nanofertilizers for improving soil fertility with a special focus on their effects on beneficial phyromicrobiome.

A multi-objective hybrid machine learning approach-based optimization for enhanced biomass and bioactive phycobiliproteins production in Nostoc sp. CCC-403.

The cyanobacterial phycobiliproteins (PBPs) are an important natural colorant for nutraceutical industries. Here, a multi-objective hybrid machine learning-based optimization approach was used for enhanced cell biomass and PBPs production simultaneously in Nostoc sp. CCC-403. A central composite design (CCD) was employed to design an experimental setup for four input parameters, including three BG-11 medium components and pH. We achieved a 61.76% increase in total PBPs production and an almost 90% increase in cell biomass by our prediction model. We also established a test genome-scale metabolic network (GSMN) for Nostoc sp. and identified potential metabolic fluxes contributing to PBPs enhanced production. This study highlights the advantage of the hybrid machine learning approach and GSMN to achieve optimization for more than one objective and serves as the foundation for future efforts to convert cyanobacteria as an economically viable source for biofuels and natural products.

High-throughput proteomics and metabolomic studies guide re-engineering of metabolic pathways in eukaryotic microalgae: A review.

Eukaryotic microalgae are a rich source of commercially important metabolites including lipids, pigments, sugars, amino acids and enzymes. However, their inherent genetic potential is usually not enough to support high level production of metabolites of interest. In order to move on from the traditional approach of improving product yields by modification of the cultivation conditions, understanding the metabolic pathways leading to the synthesis of the bioproducts of interest is crucial. Identification of new targets for strain engineering has been greatly facilitated by the rapid development of high-throughput sequencing and spectroscopic techniques discussed in this review. Despite the availability of high throughput analytical tools, examples of gathering and application of proteomic and metabolomic data for metabolic engineering of microalgae are few and mainly limited to lipid production. The present review highlights the application of contemporary proteomic and metabolomic techniques in eukaryotic microalgae for redesigning pathways for enhanced production of algal metabolites.

Molecular Analysis of Disease-Responsive Genes Revealing the Resistance Potential Against Fusarium Wilt (Fusarium udum Butler) Dependent on Genotype Variability in the Leguminous Crop Pigeonpea.

Fusarium wilt (FW), caused by Fusarium udum Butler (FU), is among the challenging factors in the production of pigeonpea. Therefore, exploring a superior pigeonpea genotype from landraces or local cultivars through the selection of innate resistance to FW using different biological and molecular approaches, and validating its resistance response, could be an alternative to sustainable crop improvement. Five distinct pigeonpea genotypes, with resistant (ICP2894) and susceptible (ICP2376) controls, were selected on the basis of the incidence percentage of FW, from three different states of India. Among them, the cultivar Richa, which displayed low incidence of FW (10.0%) during the genotype evaluation, was further examined for its innate resistance to FW. Molecular characterization of antioxidant (AO) enzyme [APX and SOD] and pathogenesis-related (PR) protein [CHS and ß-1, 3-glucanase] families were performed. The obtained results of reverse transcription-polymerase chain reaction-based expression study and in silico analysis showed a higher level of induction of PR and AO genes, and the strong interaction of their putative proteins with fungal cellobiohydrolase-c protein established their antifungal activity, conferring early plant defense responses to FU in Richa. Our study demonstrated a strong and combinatorial approach involving biological assay, molecular experiments, and in silico analysis to identify a superior pigeonpea genotype that was resistant to FW across a major biogeographic region.

Phycobiliproteins from Anabaena variabilis CCC421 and its production enhancement strategies using combinatory evolutionary algorithm approach.

The production of phycobiliproteins (PBPs) from cyanobacteria represents both the industrial application and their commercial value. In this study, the capability of Anabaena variabilis CCC421 for the production of PBPs was evaluated which was further improved by optimization of selected BG-11 medium components viz. FAC, K2HPO4 and trace metals. A design matrix approach using evolutionary algorithm comprised of genetic-algorithm (GA) and fuzzy-logic-methodology (FLM), i.e., GA-Fuzzy, was used for the optimization. The maximum production of PBPs obtained with combinatory approach of GA-Fuzzy was 408.5 mg/L at an optimum combination of factors (FAC 0.153 g/L, K2HPO4 0.2 g/L and Trace metals 0.5 ml/L) which was a 2.13 fold more than the control medium. This novel approach is very useful for modulating biological processes since various nutrients and metabolites have greater influence on these processes.

Enhancing production of microalgal biopigments through metabolic and genetic engineering.

The versatile use of biopigments in food, feed, cosmetic, pharmaceutical and analytical industries emphasized to find different and renewable sources of biopigments. Microalgae, including cyanobacteria, are becoming a potential candidate for pigment production as these have fast-growing ability, high pigment content, highly variable and also have "Generally recognized as safe" status. These algal groups are known to produce different metabolites that include hormones, vitamins, biopolythene and biochemicals. We discuss here the potential use of microalgal biopigments in our daily life as well as in food and cosmetic industries. Pigment like carotenoids has many health benefits such as antioxidant, anti-inflammatory properties and also provide photo-protection against UV radiation. This review details the effect of various abiotic and biotic factors such as temperature, light, nutrition on maximizing the pigment content in the microalgal cell. This review also highlights the potential of microalgae, whether in present native or engineered strain including the many metabolic strategies which are used or can be used to produce a higher amount of these valuable biopigments. Additionally, future challenges in the context of pigment production have also been discussed.

Cyanobacterial pigments: Perspectives and biotechnological approaches.

Cyanobacteria are the oxygenic photosynthesis performing prokaryotes and show a connecting link between plastids of eukaryotic autotrophs and prokaryotes. A variety of pigments, like chlorophyll, carotenoids and phycobiliproteins which exhibit different colors are present in cyanobacteria. Increasing consciousness about the harmful effects of synthetic or chemical dyes encouraged people to give more preference towards the usage of natural products, such as plant or microbial-derived colors in food and cosmetics. That is why cyanobacteria are exploited as a source of natural colors and have high commercial value in many industries. This review mainly focuses on different cyanobacterial pigments, their applications and modern biotechnological approaches such as genetic engineering, systems biology to enhance the production of biopigments for their potential use in pharmaceuticals, food, research, and cosmetics industries.

Impact of blue green algae (BGA) technology: an empirical evidence from northwestern Indo-Gangetic Plains.

The focus of the current study is to assess the impact of blue green algae (BGA) technology on farming practices and economic conditions of farming households. The states of Punjab, Uttar Pradesh and Haryana considered as major rice-growing states of India were chosen for the primary survey. It was observed that use of BGA resulted in 25.2% of urea reduction with an overall 3.8% increase in the yield and a marginal decrease in per acre cultivation cost. Tobit model has been used to identify the determinants of increasing cropping area under BGA. Factors such as age, education, operated land holdings and leased-in land have a positive and significant impact on area under BGA. The study also provided the comparative analysis on yield of paddy, urea consumption and income with and without BGA application. It was observed that farmers earned about 3% greater income along with 41.1% reduction in dosage of urea while reaping 1% higher yield of paddy.

Formulation of a minimal nutritional medium for enhanced lipid productivity in Chlorella sp. and Botryococcus sp. using response surface methodology.

Chlorella sp. MCC 7 and Botryococcus sp. MCC 31 were investigated to enable large-scale biodiesel production from minimal constituents in the growth medium. Response surface methodology (RSM) was used to maximise the biomass productivity and lipid yield using only nitrogen (N), phosphorus (P) and potassium (K) as urea, single super phosphate and muriate of potash. The optimum values were 0.42 g/L nitrogen; 0.14 g/L phosphorus and 0.22 g/L potassium for Chlorella sp.; and 0.46 g/L; 0.14 g/L and 0.25 g/L for Botryococcus sp. Lipid yield of 42% for Chlorella sp. and 52% in Botryococcus sp. was observed. An enhancement in lipid yield by approximately 55% for Chlorella sp. and 73% for Botryococcus sp. was registered as compared to original nutrient medium. Fourier transform infrared (FTIR) analysis of extracted lipids revealed characteristic bands for triglycerides. This study provided utilisation of a practicable nutrient recipe in the form of N, P, K input for enhanced lipid yield from the selected microalgal strains.

A comparative study reveals the higher resolution of RAPD over ARDRA for analyzing diversity of Nostoc strains.

Nostoc is a diverse genus of filamentous cyanobacteria with tremendous potential for agricultural and industrial applications. Morphometric methods and routine 16S rDNA-based identification undermines the genetic diversity and impedes strain-level differentiation. A comparative study to deduce the discriminatory power of random amplified polymorphic DNA (RAPD) and amplified ribosomal DNA restriction analysis (ARDRA) for analyzing the genetic diversity of 20 Nostoc strains of diverse geographical origin was carried out. The RAPD primer used in the study generated 100% polymorphic profile. HIP TG primer produced the highest number of bands and fragments. Five primers, viz. OPA 08, OPA 11, HIP GC, OPAH 02 and OPF 05 could produce unique bands for 11 strains. Cluster analysis using the RAPD profile showed 12.5-25% similarity among the strains. Following in silico restriction analysis, two restriction enzymes, viz. HaeIII and HinfI were selected for ARDRA. However, clustering based on the restriction pattern showed 22.5-100% similarity. Results of the present study clearly indicate higher resolution of RAPD which can be reliably used for strain-level differentiation of Nostoc strains.

Lipid production and molecular dynamics simulation for regulation of accD gene in cyanobacteria under different N and P regimes.

BACKGROUND: Microalgae grown under different nutrient deficient conditions present a good source of natural lipids with applications for several types of biofuels. The expression of acetyl-CoA carboxylase gene can further provide an insight to the mechanisms leading to enhanced lipid production under such stresses. In this study, two nutrients viz. nitrogen and phosphorus were modulated to see its effect on lipid productivity in selected cyanobacteria and its correlation with Accase followed by molecular dynamics simulation. RESULTS: Selected cyanobacteria viz. Oscillatoria sp. (SP8), Anabaena sp. (SP12), Anabaena sp. (SP13), Microcoleus sp. (SP18), and Nostoc sp. (SP20) varied in their ability to accumulate lipids which ranged from a lowest of 0.13% in Anabaena sp. (SP13) to the maximum of 7.24% in Microcoleus sp. (SP18). Microcoleus sp. (SP18) also recorded highest lipid accumulation at both N (6 mM NaNO3) and P (0.20 mM K2HPO4) limiting conditions. The overall expression of accD was found to be upregulated in both Oscillatoria sp. (SP8) and Microcoleus sp. (SP18) for all nitrogen concentrations but was differentially regulated with both positive and negative induction under phosphorus stress conditions. Maximum induction was observed in Microcoleus sp. (SP18) at 0.20 mM K2HPO4. The obtained 3D structure of SP8 protein (21.8 kDa) showed six alpha helices, while SP18 protein (16.7 kDa) exhibited four alpha helices and four beta sheets. The phi (ϕ)/psi(ψ) angles of the amino acid residues observed in Ramachandran plot analysis showed that both SP8 and SP18 proteins were highly stable with more than 90% amino acids in allowed regions. The molecular dynamics simulation results also indicated the stability of ligand-bound protein complexes. CONCLUSION: It has been demonstrated that cyanobacterial isolates are affected differently by nutrient limitation leading to variation in their lipid productivity. The same has been revealed by the behavior of accD gene expression which was regulated more by nutrients concentrations rather than the organism. However, the ligand-bound protein complexes were stable throughout MD simulations.

Effect of Nanohexaconazole on Nitrogen Fixing Blue Green Algae and Bacteria.

Nanohexaconazole is a highly efficient fungicide against Rhizoctonia solani. Nanoparticles are alleged to adversely affect the non-target organisms. In order to evaluate such concern, the present study was carried out to investigate the effect of nanohexaconazole and its commercial formulation on sensitive nitrogen fixing blue green algae (BGA) and bacteria. Various activities of algae and bacteria namely growth, N-fixation, N-assimilation, Indole acetic acid (IAA) production and phosphate solubilization were differently affected in the presence of hexaconazole. Although, there was stimulatory to slightly inhibitory effect on the growth measurable parameters of the organisms studied at the recommended dose of nanohexaconazole, but its higher dose was inhibitory to all these microorganisms. On the other hand, the recommended as well as higher dose of commercial hexaconazole showed much severe inhibition of growth and metabolic activity of these organisms as compared to the nano preparation. The uses of nanohexazconazole instead of hexaconazole as a fungicide will not only help to control various fungal pathogens but also sustain the growth and activity of these beneficial microorganisms for sustaining soil fertility and productivity.

Extraction and purification of C-phycocyanin from Spirulina platensis (CCC540).

In this study a simple protocol was developed for purifying phycocyanin (PC) from Spirulina platensis (CCC540) by using ammonium sulphate precipitation, followed by a single step chromatography by using DEAE-Cellulose-11 and acetate buffer. Precipitation with 65 % ammonium sulphate resulted in 80 % recovery of phycocyanin with purity of 1.5 (A620/A280). Thro1ugh chromatography an 80 % recovery of phycocyanin with a purity of 4.5 (A620/A280) was achieved. In SDS_PAGE analysis, the purified PC showed the presence of two subunit α (16 kD) and ß (17 kD).

Protocol optimization for enhanced production of pigments in Spirulina.

Spirulina has attracted special attention due to its importance as human foodstuff and natural colours with specific functional properties. These functional properties have been attributed to phycobilins, carotenoids, phenolics and unsaturated fatty acids. Present study was conducted under controlled phytotron conditions to identify the efficient strains of Spirulina in terms of pigment synthesis and to optimize their enhanced production. Methodology for enhanced production was standardized by varying specific environmental parameters (light intensity, temperature, carbon dioxide concentration, pH and NaCl level). Different strains of Spirulina depicted variability and environmental parameters showed distinct influence on pigments. Growth and pigment production was recorded to be most efficient under optimized conditions of light intensity (70 µmol m-2 s-1), temperature (30 °C), CO2 concentration (550 ppm and 750 ppm), pH (10.5) and NaCl level (2 g L-1).