A novel integrated approach employing Desertifilum tharense BERC-3 for efficient wastewater valorization and recycling for developing peri-urban algae farming system.

Peri-urban environments are significant reservoirs of wastewater, and releasing this untreated wastewater from these resources poses severe environmental and ecological threats. Wastewater mitigation through sustainable approaches is an emerging area of interest. Algae offers a promising strategy for carbon-neutral valorization and recycling of urban wastewater. Aiming to provide a proof-of-concept for complete valorization and recycling of urban wastewater in a peri-urban environment in a closed loop system, a newly isolated biocrust-forming cyanobacterium Desertifilum tharense BERC-3 was evaluated. Here, the highest growth and lipids productivity were achieved in urban wastewater compared to BG11 and synthetic wastewater. D. tharense BERC-3 showed 60-95% resource recovery efficiency and decreased total dissolved solids, chemical oxygen demand, biological oxygen demand, nitrate nitrogen, ammonia nitrogen and total phosphorus contents of the water by 60.37%, 81.11%, 82.75%, 87.91%, 85.13%, 85.41%, 95.87%, respectively, making it fit for agriculture as per WHO's safety limits. Soil supplementation with 2% wastewater-cultivated algae as a soil amender, along with its irrigation with post-treated wastewater, improved the nitrogen content and microbial activity of the soil by 0.3-2.0-fold and 0.5-fold, respectively. Besides, the availability of phosphorus was also improved by 1.66-fold. The complete bioprocessing pipeline offered a complete biomass utilization. This study demonstrated the first proof-of-concept of integrating resource recovery and resource recycling using cyanobacteria to develop a peri-urban algae farming system. This can lead to establishing wastewater-driven algae cultivation systems as novel enterprises for rural migrants moving to urban areas.

Characterization of a newly isolated self-flocculating microalga Bracteacoccus pseudominor BERC09 and its evaluation as a candidate for a multiproduct algal biorefinery.

Microalgae have the highest capability to fix the atmospheric carbon and wastewater-derived nutrients to produce high-value bioproducts including lipids and carotenoids. However, their lower titers and single-product-oriented biomass processing have made the overall process expensive. Hence, increased metabolite titer and processing of the biomass for more than one product are required to ensure the commercial robustness of the algal biorefinery. In this study, a newly isolated algal strain was identified as Bracteacoccus pseudominor BERC09 through phylogenetic analysis based on the 18S rRNA gene sequence. Basic characterization of the strain revealed its promising potential to produce carotenoids and lipids. The lipids and carotenoid biosynthesis pathways of BERC09 were further triggered by manipulating the abiotic factors including nitrogen sources (NaNO3, KNO3, NH4Cl, Urea), nitrogen concentrations (0.06-0.36 gL-1), light intensity (150 µmolm-2s-1 to 300 µmolm-2s-1), and light quality (white and blue). Resultantly, 300 µmolm-2s-1 of blue light yielded 0.768 gL-1 of biomass, 8.4 mgg-1 of carotenoids, and 390 mgg-1 of lipids, and supplementation of 0.36 gL-1 of KNO3 further improved metabolism and yielded 0.814 gL-1 of biomass, 11.86 mgg-1 of carotenoids, and 424 mgg-1 of lipids. Overall, the optimal combination of light and nitrogen concurrently improved biomass, carotenoids, and lipids by 3.5-fold, 6-fold, and 4-fold than control, respectively. Besides, the excellent glycoproteins-based self-flocculation ability of the strain rendered an easier harvesting via gravity sedimentation. Hence, this biomass can be processed in a cascading fashion to use this strain as a candidate for a multiproduct biorefinery to achieve commercial robustness and environmental sustainability.

Engineering the metabolic pathways of lipid biosynthesis to develop robust microalgal strains for biodiesel production.

Algal lipids have shown promising feedstock to produce biodiesel due to higher energy content, higher cetane number, and renewable nature. However, at present, the lipid productivity is too low to meet the commercial needs. Various approaches can be employed to enhance the lipid content and lipid productivity in microalgae. Stress manipulation is an attractive option to modify the algal lipid content, but it faces the drawback of time-consuming production processing and lack of information about molecular mechanisms related to triacylglycerides production in response to stress. Developing the robust hyper lipid accumulating algal strains has gained momentum due to advances in metabolic engineering and synthetic biology tools. Understanding the molecular basis of lipid biosynthesis followed by reorienting the related pathways through genomic modification is an alluring strategy that is believed to achieve the industrial and economic robustness. This review portrays the use of integrated OMIC approaches to elucidate the molecular mechanisms of strain adaptability in response to stress conditions, and identification of molecular pathways that should become novel targets to develop novel algal strains. Moreover, an update on the metabolic engineering approaches to improve the lipid production in microalgae is also provided.

State-of-the-Art Genetic Modalities to Engineer Cyanobacteria for Sustainable Biosynthesis of Biofuel and Fine-Chemicals to Meet Bio-Economy Challenges.

In recent years, metabolic engineering of microorganisms has attained much research interest to produce biofuels and industrially pertinent chemicals. Owing to the relatively fast growth rate, genetic malleability, and carbon neutral production process, cyanobacteria has been recognized as a specialized microorganism with a significant biotechnological perspective. Metabolically engineering cyanobacterial strains have shown great potential for the photosynthetic production of an array of valuable native or non-native chemicals and metabolites with profound agricultural and pharmaceutical significance using CO2 as a building block. In recent years, substantial improvements in developing and introducing novel and efficient genetic tools such as genome-scale modeling, high throughput omics analyses, synthetic/system biology tools, metabolic flux analysis and clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease (CRISPR/cas) systems have been made for engineering cyanobacterial strains. Use of these tools and technologies has led to a greater understanding of the host metabolism, as well as endogenous and heterologous carbon regulation mechanisms which consequently results in the expansion of maximum productive ability and biochemical diversity. This review summarizes recent advances in engineering cyanobacteria to produce biofuel and industrially relevant fine chemicals of high interest. Moreover, the development and applications of cutting-edge toolboxes such as the CRISPR-cas9 system, synthetic biology, high-throughput "omics", and metabolic flux analysis to engineer cyanobacteria for large-scale cultivation are also discussed.

Mitochondrial gene cytochrome c oxidase I (CO1) used for molecular identification of Bactrocera zonata in Pakistan.

Bactrocera zonata is fruit pest mostly attacked on peach and cause heavy destruction in production of peach fruits by sucking their juice. For their management, we start to detect them on basis of their molecular characterization. As mitochondrial genome encodes a gene COI used as biomarker for identification of eukaryotes including insects. In present study, we amplified COI gene and cloned into pTZ57R/T vector (Fermentas). Cloned gene was confirmed through restriction analysis and sequenced through its entirety on both strands from Macrogen (South Korea) by Sanger sequencing method. Different computational tools were utilized for comparative analysis of sequence with other related sequences retrieved from databases. Related species were identified through phylogenetic analysis using Mega 7 tool. Pairwise sequence alignment showed the sequence identity about 96% with Bactrocera zonata. By identifying the pests with more authentic molecular biomarker may help the research to control them more effectively in future.

Omics Technologies for Microalgae-based Fuels and Chemicals: Challenges and Opportunities.

BACKGROUND: Microalgae have been suggested as promising feedstocks of significant biotechnological interest due to their enormous potential for the sustainable production of industrially valuable compounds such as lipids/fatty acids, proteins, metabolites, pigments, and biofuels. However, exploitation of algal biomass for commercial purposes is still in its infancy due to the dearth of the knowledge regarding state-of-the-art sophisticated technologies. OBJECTIVES: The main objective of the study was to review the explosions of innovative strategies that biological sciences have witnessed over the past several years, enabling the scientific community and research-based organizations to scrutinize entire classes of biomolecules from a cell type or whole organism, collectively titled as 'Omics, including genomics, transcriptomics, proteomics and lipidomics. METHODS: An effort has been made to analyze the relative advantages and drawbacks of these technologies by reviewing and consulting the available literature. RESULTS: Nevertheless, these 'Omics strategies have played a key role in the better understanding of the underlying mechanisms of specific gene products, regulatory constraints, metabolic pathways and potential genes and protein targets which might be employed in tailoring microalgae for enhanced productivity. In this critical review, an understanding of recent omics technologies, their potential applications, and limitations for microalgae-based fuel and other commodity chemicals are comprehensively discussed. CONCLUSION: In the scenario of uncertain petro-based reserves, global warming and energy insecurity, the exploration of metabolic potentialities of microalgae using "Omics" are believed to be a green and environmentally-responsive approach that will further expand its industrial and environmental scope.