Evaluation of fuel properties for possible biodiesel output based on the fatty acid composition of oleaginous plants and microalgae.

In the past decade, there has been a significant rise in sustainable biomass based biofuel production to address energy needs while mitigating environmental impacts. Traditionally, bioethanol was used for biofuel production, but concerns over food security and environmental preservation have led to growing interest in alternative sources such as neutral lipids from vegetable oil and microalgae for biodiesel production. This research paper evaluates the potential of various oleaginous plants and microalgae as feedstocks for biodiesel production, with a focus on their fatty acid composition and its impact on biodiesel properties. The study examines the fatty acid profiles of 43 different plant and microalgae species and employs various equations to estimate key physical properties of biodiesel. Additionally, the communication compares these properties to International Biodiesel Standards (EN 14214 and ASTM D6751-08) to assess the suitability of the derived biodiesel for commercial use. It is impossible to describe a single composition that is optimal in terms of all essential fuel properties due to the opposing effects of some structural features of the Fatty Acid Methyl Esters (FAME). However, biodiesel should contain relatively low concentrations of both long chain saturated and polyunsaturated FAME to ensure adequate efficiency in terms of low temperature operability and oxidative stability. The results reveal significant variations in properties amongst different feedstocks, highlighting the importance of feedstock selection in biodiesel production. The study also establishes correlations between various fuel properties, providing valuable insights in to optimizing biodiesel production processes, which will be of great use to researchers, engineers, and stakeholders involved in biodiesel production.

Elevated inorganic carbon and salinity enhances photosynthesis and ATP synthesis in picoalga Picocystis salinarum as revealed by label free quantitative proteomics.

Saline soda lakes are of immense ecological value as they niche some of the most exclusive haloalkaliphilic communities dominated by bacterial and archaeal domains, with few eukaryotic algal representatives. A handful reports describe Picocystis as a key primary producer with great production rates in extremely saline alkaline habitats. An extremely haloalkaliphilic picoalgal strain, Picocystis salinarum SLJS6 isolated from hypersaline soda lake Sambhar, Rajasthan, India, grew robustly in an enriched soda lake medium containing mainly Na2CO3, 50 g/l; NaHCO3, 50 g/l, NaCl, 50 g/l (salinity ≈150‰) at pH 10. To elucidate the molecular basis of such adaptation to high inorganic carbon and NaCl concentrations, a high-throughput label-free quantitation based quantitative proteomics approach was applied. Out of the total 383 proteins identified in treated samples, 225 were differentially abundant proteins (DAPs), of which 150 were statistically significant (p < 0.05) including 70 upregulated and 64 downregulated proteins after 3 days of growth in highly saline-alkaline medium. Most DAPs were involved in photosynthesis, oxidative phosphorylation, glucose metabolism and ribosomal structural components envisaging that photosynthesis and ATP synthesis were central to the salinity-alkalinity response. Key components of photosynthetic machinery like photosystem reaction centres, adenosine triphosphate (ATP) synthase ATP, Rubisco, Fructose-1,6-bisphosphatase, Fructose-bisphosphate aldolase were highly upregulated. Enzymes peptidylprolyl isomerases (PPIase), important for correct protein folding showed remarkable marked-up regulation along with other chaperon proteins indicating their role in osmotic adaptation. Enhanced photosynthetic activity exhibited by P. salinarum in highly saline-alkaline condition is noteworthy as photosynthesis is suppressed under hyperosmotic conditions in most photosynthetic organisms. The study provided the first insights into the proteome of extremophilic alga P. salinarum exhibiting extraordinary osmotic adaptation and proliferation in polyextreme conditions prevailing in saline sodic ecosystems, potentially unraveling the basis of resilience in this not so known organism and paves the way for a promising future candidate for biotechnological applications and model organism for deciphering the molecular mechanisms of osmotic adaptation. The mass spectrometry proteomics data is available at the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD037170.

Significance of Plant Growth Promoting Rhizobacteria in Grain Legumes: Growth Promotion and Crop Production.

Grain legumes are an important component of sustainable agri-food systems. They establish symbiotic association with rhizobia and arbuscular mycorrhizal fungi, thus reducing the use of chemical fertilizers. Several other free-living microbial communities (PGPR-plant growth promoting rhizobacteria) residing in the soil-root interface are also known to influence biogeochemical cycles and improve legume productivity. The growth and function of these microorganisms are affected by root exudate molecules secreted in the rhizosphere region. PGPRs produce the chemicals which stimulate growth and functions of leguminous crops at different growth stages. They promote plant growth by nitrogen fixation, solubilization as well as mineralization of phosphorus, and production of phytohormone(s). The co-inoculation of PGPRs along with rhizobia has shown to enhance nodulation and symbiotic interaction. The recent molecular tools are helpful to understand and predict the establishment and function of PGPRs and plant response. In this review, we provide an overview of various growth promoting mechanisms of PGPR inoculations in the production of leguminous crops.

Biochemical parameters and 18S rRNA gene sequence analysis amongst green microalgal strains from selected aquatic sites of Eastern India.

In the present study, 24 green microalgae strains were isolated from selected aquatic sites of India. These were microscopically identified as Chlamydomonas sp., Scenedesmus sp., Chlorella sp., Dictyosphaerium sp. and Dunaliella sp. Nannochloropsis sp. (MCC 25), was used as a reference strain. Results showed that Dictyosphaerium sp. (MCC 10 and MCC 12) showed relatively higher nutritive content. The total soluble proteins in the reference strain was 21.4%, whereas it showed carbohydrate content of 17.2% and the lipids were 3.4% on a dry weight basis. Best performing strains were identified by biochemical characterization. Five genera were selected for molecular identification since they were the most representative based upon their area of isolation and their optimum content of total soluble proteins, carbohydrates and lipids. 18S rRNA sequencing authenticated their identification as Scenedesmus sp., Dictyosphaerium sp. and Chlorella sp. The sequences of these have been submitted in NCBI database with accession numbers as KT808247-KT808251. The correlation matrix showed positive correlation between carbohydrates and lipids, while negative correlation was seen between proteins and carbohydrates and between proteins and lipids. This study emphasizes the need for complete compositional analysis of the biomass for the possible applicability in the area of value addition.

Influence of nutrient formulations on growth, lipid yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp.

The study was conducted to analyse the influence of three nutrient formulations, namely BG-11 medium, BBM and TAP medium, on growth potential and lipid yield of two microalgal genera (Botryococcus sp. and Chlorella sp.) and to study the roles of N, P and other major nutrients. The study focussed on the general patterns of starch and lipid synthesis and storage and to further assess how photosynthetic carbon partitioning into starch and lipid is altered by conditions in growth media such as N and C presence as seen in BG11 medium which are known to induce neutral lipid production and the lack of it in BBM and TAP medium. BG-11 medium performed better as compared to BBM and TAP medium in terms of biomass productivity and lipid yield. The lipid yield was highest in Botryococcus sp. (63.03% dry wt.) and Chlorella sp. (50.27% dry wt.) at 30th day of incubation. Mean biomass productivity was highest for Botryococcus in BBM medium (6.14 mg/L/day) and for Chlorella in BG-11 medium (4.97 mg/L/day). Mean lipid productivity (50.78% and 39.36%) was highest in BG11 medium for both Botryococcus and Chlorella species, respectively. A sharp decline in sugar content was observed in the late stationary phase of growth from 30th day to 45th day. Fatty acid methyl ester (FAME) profile of the extracted lipids showed predominantly oleic acid, followed by palmitic acid and stearic acid in both the strains when grown in BG-11 medium. The other biodiesel quality parameters were in accordance with the international standards. A complex relationship was found between chemical composition and biodiesel properties. Proximity analysis indicated that the fuel properties of biodiesels are determined by a number of parameters and by the combination of different chemical compositions. The results provide an insight into organic carbon partitioning into lipid compounds and how the organism's lipid metabolism changes due to N-deplete culturing in TAP medium and inorganic carbon source availability as seen in BG-11 and BBM medium.

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.

Assessing biodiesel quality parameters for wastewater grown Chlorella sp.

Microalgae are reported as the efficient source of renewable biodiesel which should be able to meet the global demand of transport fuels. Present study is focused on assessment of wastewater grown indigenous microalga Chlorella sp. for fuel quality parameters. This was successfully grown in secondary treated waste water diluted with tap water (25% dilution) in glass house. The microalga showed a dry weight of 0.849 g L-1 with lipid content of 27.1% on dry weight basis on 21st day of incubation. After transesterification, the yield of fatty acid methyl ester was 80.64% with major fatty acids as palmitic, linoleic, oleic and linolenic. The physical parameters predicted from empirical equations in the biodiesel showed cetane number as 56.5, iodine value of 75.5 g I2 100 g-1, high heating value 40.1 MJ kg-1, flash point 135 °C, kinematic viscosity 4.05 mm2 s-1 with density of 0.86 g cm3 and cold filter plugging point as 0.7 °C. Fourier transform infra-red (FTIR), 1H, 13C NMR spectrum confirmed the chemical nature of biodiesel. The results indicated that the quality of biodiesel was almost as per the criterion of ASTM standards; hence, wastewater grown Chlorella sp. can be used as a promising strain for biodiesel production.

Influence of light intensity, temperature and CO2 concentration on growth and lipids in green algae and cyanobacteria.

Effects of the environmental variables such as light intensity (Vmol photons m⁻² S⁻¹), temperature (*C) and CO2 concentration (ppm) on chlorophyll, total soluble proteins and lipids were studied in selected microalgal strains from Chlorophyceae (Chlamydomonas sp., Scenedesmus sp., Chlorella sp., Kirchneriella sp.) and cyanobacteria (Nostoc sp.1, Anabaena sp., Nostoc sp. 2, Cylindrospermum sp.). Cultures were grown under controlled conditions at the National Phytotron Facility, Indian Agricultural Research Institute (IARI), New Delhi. Our results showed that chlorophyll concentration enhanced with increased C02. Chlorella exhibited the highest chlorophyll at 850 ppm CO2 and 28*C; for Chlamydomonas it was at 78 µmol photons m⁻² S⁻¹ light intensity. In Cylindrospernum, total soluble proteins decreased with enhanced C02, and were highest at 18*C. In Anabaena, a light intensity of 65 µmol photons m⁻² S⁻¹ was best for maximum total soluble proteins. In Chlorella, CO2 @ 850 ppm was most suited for maximum lipid accumulation. In Kirchneriella, increase in temperature, from 18*C up to 370C, increased total lipids; the highest was at 28'C. In Chlamydomonas, the light intensity of 78 µmol photons m⁻² S⁻¹ was optimum for lipid accumulation and the maximum total lipids was 30.8 (% dry wt.).

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).

Bioprospecting and indexing the microalgal diversity of different ecological habitats of India.

Our study reports the collection, biodiversity analyses, isolation and identification of microalgae from different habitats of India. Cyanophyceae and Chlorophyceae were the most dominant algal groups recorded, with the highest number being recorded for non-heterocystous cyanobacteria (48), followed by 44 unicellular forms. Sagar Island, Sunderbans recorded the greatest number of algae, and unicellular/colonial green algae were present in all the samples. Shannon's Diversity Index was highest in Koikhali, Sunderbans, followed by Rushikulya River, Odisha. Selective enrichment, purification through serial dilution followed by plating and regular observations led to the isolation of sixteen strains. Identification was done by using microscopic observations, supported with standard monographs and classified as belonging to seven genera (Chlorella, Chlorococcum, Kirchneria, Scenedesmus, Chlamydomonas, Tetracystis and Ulothrix). 18S rDNA sequencing was undertaken for four strains. The set of sixteen strains were screened under standard cultural conditions for their growth kinetics and Chlorella sorokiniana MIC-G5, followed by Chlorella sp. MIC-G4 exhibited the highest growth rates. The strain Chlorococcum sp. MIC-G2 recorded highest chlorophyll, while MIC-G3 ranked highest for carbohydrates. The study aided in identifying the dominant microalgae in the diverse habitats and characterizing their growth rate and carbohydrate content, providing a valuable germplasm for further utilization in agriculture and industry.

Biochemical modulation of growth, lipid quality and productivity in mixotrophic cultures of Chlorella sorokiniana.

The genus Chlorella is a widely employed microalga for biodiesel, as it can be grown using photo/mixo/heterotrophic mode of cultivation. The present investigation was undertaken with the hypothesis that addition of different substrates (amino acids, carbon sources, vitamins) along with reducing agents may aid in diverting Acetyl CoA to malonyl CoA or fatty acid biosynthesis, under mixotrophic conditions in Chlorella sorokiniana. Preliminary investigations undertaken with two reducing agents individually (sodium thiosulphate and methyl viologen) along with selected substrates revealed the promise of sodium thiosulphate (1%) in enhancing lipid accumulation significantly. Further, the role of inclusion of twelve substrates and sodium thiosulphate revealed that supplementation with tryptophan (0.1%) recorded 57.28% enhancement in lipid productivity on 4(th) day. Highest values of lipid productivity of 33% were recorded on 8(th) day in 0.1% glucose supplemented medium containing sodium thiosulphate. Fatty Acid Methyl Ester (FAME) profiles generated revealed significant reduction in the content of Poly unsaturated fatty acids (PUFA) and enhanced Mono unsaturated fatty acids (MUFA) (especially oleic acid) in the treatments involving tryptophan, Vitamin B12, sodium pyruvate and glucose. This study reveals the promise of using sodium thiosulphate along with selected substrate for enriching the quality and quantity of lipids, which can be valuable for exploiting algae as a source of biodiesel.

Physiological studies on endorhizospheric establishment of Azotobacter chroococcum in wheat.

Ten strains of Azotobacter chroococcum were studied for their ability to invade the endorhizosphere of wheat. Strain W-5 exhibited ability to invade endorhizosphere as shown in the microscopic observations. This strain was compared with the strain OA-3 which did not invade the endorhizosphere zone. Strain W-5 showed higher production of cellulase and pectinase than OA-3. Both the strains induced defense enzymes in the host plant. However, induction of peroxidase and phenylalanine ammonia lyase activities (PAL) was higher in OA-3 than W-5. Quantitative differences in flavonoid like compounds obtained from root extracts and root exudates of plants inoculated with these strains were observed.

Induction of salt tolerance in Azolla microphylla Kaulf through modulation of antioxidant enzymes and ion transport.

Azolla microphylla plants exposed directly to NaCl (13 dsm(-1)) did not survive the salinity treatment beyond a period of one day, whereas plants exposed directly to 4 and 9 dsm(-1) NaCl were able to grow and produce biomass. However, plants pre-exposed to NaCl (2 dsm(-1)) for 7 days on subsequent exposure to 13 dsm(-1) NaCl were able to grow and produce biomass although at a slow rate and are hereinafter designated as pre-exposed plants. The pre-exposed and directly exposed plants distinctly differed in their response to salt in terms of lipid peroxidation, proline accumulation, activity of antioxidant enzymes, such as SOD, APX, and CAT, and Na(+)/K(+) ratio. Efficient modulation of antioxidant enzymes coupled with regulation of ion transport play an important role in the induction of salt tolerance. Results show that it is possible to induce salt adaptation in A. microphylla by pre-exposing them to low concentrations of NaCl.

Isolation and characterisation of phycobiliprotein rich mutant of cyanobacterium Synechocystis sp.

Chemical (N' methyl-N'-nitro-N-nitrosoguanidine) mutagenesis and penicillin selection were utilised to isolate a phenotypically altered mutant of cyanobacterium Synechocystis sp. This mutant (ntm60A) exhibited enhanced protein content and nitrogen fixing potential but lower amount of chlorophyll and nitrate reductase activity. A remarkable and significant increase was observed in the total phycobiliprotein content of the mutant, especially in relation to the amount of phycoerythrin. This strain can be exploited as a rich source of natural coloring agent such as phycobilins in the bioindustry.