Browsing by Author "Puri M"

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Bioprospecting Indigenous Marine Microalgae for Polyunsaturated Fatty Acids Under Different Media Conditions
(Frontiers Media S A, 2022-03-17) Jain P; Minhas AK; Shukla S; Puri M; Barrow CJ; Mandal S; Khanna N
Marine microalgae produce a number of valuable compounds that have significant roles in the pharmaceutical, biomedical, nutraceutical, and food industries. Although there are numerous microalgal germplasms available in the marine ecosystem, only a small number of strains have been recognized for their commercial potential. In this study, several indigenous microalgal strains were isolated from the coast of the Arabian Sea for exploring the presence and production of high-value compounds such as polyunsaturated fatty acids (PUFAs). PUFAs are essential fatty acids with multiple health benefits. Based on their high PUFA content, two isolated strains were identified by ITS sequencing and selected for further studies to enhance PUFAs. From molecular analysis, it was found both the strains were green microalgae: one of them was a Chlorella sp., while the other was a Planophila sp. The two isolated strains, together with a control strain known for yielding high levels of PUFAs, Nannochloropsis oculata, were grown in three different nutrient media for PUFA augmentation. The relative content of α-linolenic acid (ALA) as a percentage of total fatty acids reached a maximum of 50, 36, and 50%, respectively, in Chlorella sp., Planophila sp., and N. oculata. To the best of our knowledge, this is the first study in exploring fatty acids in Planophila sp. The obtained results showed a higher PUFA content, particularly α-linolenic acid at low nutrients in media.
Development of sustainable downstream processing for nutritional oil production
(Frontiers Media SA, 2023-10-10) Rollin S; Gupta A; Franco CMM; Singh S; Puri M; Sforza E
Nutritional oils (mainly omega-3 fatty acids) are receiving increased attention as critical supplementary compounds for the improvement and maintenance of human health and wellbeing. However, the predominant sources of these oils have historically shown numerous limitations relating to desirability and sustainability; hence the crucial focus is now on developing smarter, greener, and more environmentally favourable alternatives. This study was undertaken to consider and assess the numerous prevailing and emerging techniques implicated across the stages of fatty acid downstream processing. A structured and critical comparison of the major classes of disruption methodology (physical, chemical, thermal, and biological) is presented, with discussion and consideration of the viability of new extraction techniques. Owing to a greater desire for sustainable industrial practices, and a desperate need to make nutritional oils more available; great emphasis has been placed on the discovery and adoption of highly sought-after ‘green’ alternatives, which demonstrate improved efficiency and reduced toxicity compared to conventional practices. Based on these findings, this review also advocates new forays into application of novel nanomaterials in fatty acid separation to improve the sustainability of nutritional oil downstream processing. In summary, this review provides a detailed overview of the current and developing landscape of nutritional oil; and concludes that adoption and refinement of these sustainable alternatives could promptly allow for development of a more complete ‘green’ process for nutritional oil extraction; allowing us to better meet worldwide needs without costing the environment.
Harnessing the evolutionary information on oxygen binding proteins through Support Vector Machines based modules
(BioMed Central Ltd, 2018-12-01) Muthukrishnan S; Puri M
Objectives: The arrival of free oxygen on the globe, aerobic life is becoming possible. However, it has become very clear that the oxygen binding proteins are widespread in the biosphere and are found in all groups of organisms, including prokaryotes, eukaryotes as well as in fungi, plants, and animals. The exponential growth and availability of fresh annotated protein sequences in the databases motivated us to develop an improved version of "Oxypred" for identifying oxygen-binding proteins. Results: In this study, we have proposed a method for identifying oxy-proteins with two different sequence similarity cutoffs 50 and 90%. A different amino acid composition based Support Vector Machines models was developed, including the evolutionary profiles in the form position-specific scoring matrix (PSSM). The fivefold cross-validation techniques were applied to evaluate the prediction performance. Also, we compared with existing methods, which shows nearly 97% recognition, but, our newly developed models were able to recognize almost 99.99 and 100% in both oxy-50 and 90% similarity models respectively. Our result shows that our approaches are faster and achieve a better prediction performance over the existing methods. The web-server Oxypred2 was developed for an alternative method for identifying oxy-proteins with more additional modules including PSSM, available at http://bioinfo.imtech.res.in/servers/muthu/oxypred2/home.html.
Harvesting optimization and Omega-3 recovery improvement from Schizochytrium DT3 using surfactant-aided dispersed air flotation: Response surface methodology
(Elsevier B.V., 2024-06) Alhattab M; Puri M
Microalgae's potential to produce high value bioactives is contingent on the cost-effective harvesting of algal biomass. The use of CTAB (cetyl trimethylammonium bromide) assisted dispersed air flotation as a harvesting technique for Schizochytrium biomass, was optimized as a cost-effective means for recovery (R) and concentration factor (CF) using Response Surface Methodology. To the best of our knowledge, this is the first study to employ surfactant assisted dispersed air flotation (SDAF) in the recovery of a heterotrophic thraustochytrid strain. A Box-Behnken design of experiment investigating the operating parameters of CTAB amount, air flow rate and volume on Schizochytrium was employed. Initially, both responses were analyzed individually and then used to simultaneously maximize both variables. The optimized conditions of CTAB (500 mg/L), air flow of 2 L/min and volume of 600 mL resulted in a R of 91 % and CF of 19 times. Although a secondary step is necessary for further concentration, this technique utilizes 70 times less energy as compared to conventional centrifugation techniques which are used to recover Schizochytrium sp. This is significant as this technique can be easily adapted to existing bioreactors, as they are already equipped with gas spargers. In addition, the presence of surfactant carried through demonstrated an improvement in the recovery of long chain poly unsaturated fatty acid (PUFA) by 6 %, particularly in DHA and DPA which was not observed in washed biomass.
Integration of the Exogenous Tuning of Thraustochytrid Fermentation and Sulfur Polymerization of Single-Cell Oil for Developing Plant-like Oils
(MDPI (Basel, Switzerland), 2022-10-01) Gupta A; Worthington MJH; Chalker JM; Puri M; Shene C
In this study, we have demonstrated a bioprocessing approach encompassing the exogenous addition of low-molecular-weight compounds to tune the fatty acid (FA) profile in a novel thraustochytrid strain to produce desirable FAs. Maximum lipid recovery (38%, dry wt. biomass) was obtained at 1% Tween 80 and 0.25 mg/L of Vitamin B12. The transesterified lipid showed palmitic acid (C16, 35.7% TFA), stearic acid (C18, 2.1% TFA), and oleic acid (C18:1, 18.7% TFA) as the main components of total FAs, which are mainly present in plant oils. Strikingly, D-limonene addition in the fermentation medium repressed the production of polyunsaturated fatty acid (PUFAs). Sulfur-polymerization-guided lipid separation revealed the presence of saturated (SFAs, 53% TFA) and monounsaturated fatty acids (MUFAs, 46.6% TFA) in thraustochytrid oil that mimics plant-oil-like FA profiles. This work is industrially valuable and advocates the use of sulfur polymerization for preparation of plant-like oils through tuneable thraustochytrid lipids.
Marine-Derived Lipases for Enhancing Enrichment of Very-Long-Chain Polyunsaturated Fatty Acids with Reference to Omega-3 Fatty Acids
(MDPI (Basel, Switzerland), 2024-07-01) Karia M; Kaspal M; Alhattab M; Puri M
Omega-3 fatty acids are essential fatty acids that are not synthesised by the human body and have been linked with the prevention of chronic illnesses such as cardiovascular and neurodegenerative diseases. However, the current dietary habits of the majority of the population include lower omega-3 content compared to omega-6, which does not promote good health. To overcome this, pharmaceutical and nutraceutical companies aim to produce omega-3-fortified foods. For this purpose, various approaches have been employed to obtain omega-3 concentrates from sources such as fish and algal oil with higher amounts of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Among these techniques, enzymatic enrichment using lipase enzymes has gained tremendous interest as it is low in capital cost and simple in operation. Microorganism-derived lipases are preferred as they are easily produced due to their higher growth rate, and they hold the ability to be manipulated using genetic modification. This review aims to highlight the recent studies that have been carried out using marine lipases for the enrichment of omega-3, to provide insight into future directions. Overall, the covalent bond-based lipase immobilization to various support materials appears most promising; however, greener and less expensive options need to be strengthened.
Mutagenesis treatment of Mortierella alpina for PUFA production enhancement for future food development
(Elsevier B.V., 2025-06) Alhattab M; Lebeau J; Singh S; Puri M
Random mutagenesis has been identified as a key tool for improving microbial and fungal strains enabling the development of isolates with improved traits suited for industrial scale metabolite production to enhance the nutritional value of future foods. Presented here, is a random mutagenesis strategy employed to assess the effect of 5-fluorouracil (20-200 µg/ml), alone and in combination with the secondary agents octyl gallate and nocodazole, and diethyl sulfate (0.1 to 1 %) chemical mutagenic agents, on the biomass and lipid production as well as the FAME profile. Interestingly, a correlation was demonstrated between 5-fluorouracil exposure time and the arachidonic acid content, which was also influenced by the concentration used. 5-fluororuracil of 100 µg/ml treatment for 48 h resulted in the highest arachidonic acid (% TFA) content in isolates. Mutant M5F047 isolated with 5-fluororuracil (100 µg/ml) alone, proved to be most superior in terms of polyunsaturated fatty acid (PUFA) and arachidonic acid production, as compared to the Mortierella alpina wild type strain, with enhancements that doubled that of the parent strain. These improvements are more favorable for industrial scale production of arachidonic acid, a precursor of meaty flavour to improve plant-based meats in future food development.
Oleaginous Microbial Lipids’ Potential in the Prevention and Treatment of Neurological Disorders
(MDPI (Basel, Switzerland), 2024-02-06) Alhattab M; Moorthy LS; Patel D; Franco CMM; Puri M; Zhang T
The products of oleaginous microbes, primarily lipids, have gained tremendous attention for their health benefits in food-based applications as supplements. However, this emerging biotechnology also offers a neuroprotective treatment/management potential for various diseases that are seldom discussed. Essential fatty acids, such as DHA, are known to make up the majority of brain phospholipid membranes and are integral to cognitive function, which forms an important defense against Alzheimer’s disease. Omega-3 polyunsaturated fatty acids have also been shown to reduce recurrent epilepsy seizures and have been used in brain cancer therapies. The ratio of omega-3 to omega-6 PUFAs is essential in maintaining physiological function. Furthermore, lipids have also been employed as an effective vehicle to deliver drugs for the treatment of diseases. Lipid nanoparticle technology, used in pharmaceuticals and cosmeceuticals, has recently emerged as a biocompatible, biodegradable, low-toxicity, and high-stability means for drug delivery to address the drawbacks associated with traditional medicine delivery methods. This review aims to highlight the dual benefit that lipids offer in maintaining good health for disease prevention and in the treatment of neurological diseases.
Opportunities for agri-food chains to become energy-smart
(FAO, 2015-11) Sims REH; Flammini A; Puri M; Bracco S
Production of Alginate Oligosaccharides (AOSs) Using Enhanced Physicochemical Properties of Immobilized Alginate Lyase for Industrial Application
(MDPI (Basel, Switzerland), 2024-03-04) Kaur S; Abraham RE; Franco CMM; Puri M; Kumagai Y; Kishimura H; Zhu B; Rahman A
Alginate lyase (AL) is a polysaccharide-degrading enzyme that can degrade alginate by hydrolyzing glycosidic bonds and produces unsaturated alginate oligosaccharides (AOSs). These AOSs have wide therapeutic and nutraceutical applications. However, to produce alginate oligosaccharides in a cost-effective manner is challenging due to the low availability and high cost of this degrading enzyme. Immobilization of the enzyme facilitates industrial applications owing to its stability, reusability, and cost-effectiveness. This study was focused on the enhancement of the properties of alginate lyase and improvement of the production of AOS. Alginate lyase was immobilized on magnetic nanoparticles (NPs) using glutaraldehyde as the crosslinker. The study showed that the maximum binding achieved between NPs and protein in the enzyme was 71% at a ratio of 1:150 NP:protein. As a result of immobilization, the optimum activity of free enzyme which was obtained at 37 °C and pH 7.4 changed to 45 °C and pH 9. Furthermore, the enzyme was thermostable at 45 °C for 3 h with up to 50% reusability for six consecutive cycles. Storage stability after 15 days showed ~67% relative hydrolysis of alginate. The free alginate lyase (25 IU) showed 76% raw biomass (seaweed) hydrolysis which is higher compared to 63% provided by the immobilized enzyme. As a result of efficient hydrolysis, AOSs with molecular weight profile of 370–1040 kDa were produced and detected using HPLC.
Purification and Characterization of a Novel Alginate Lyase from a Marine Streptomyces Species Isolated from Seaweed
(MDPI (Basel, Switzerland), 2021-11-01) Nguyen TNT; Chataway T; Araujo R; Puri M; Franco CMM
Alginate, a natural polysaccharide derived from brown seaweed, is finding multiple applications in biomedicine via its transformation through chemical, physical, and, increasingly, enzymatic processes. In this study a novel alginate lyase, AlyDS44, was purified and characterized from a marine actinobacterium, Streptomyces luridiscabiei, which was isolated from decomposing seaweed. The purified enzyme had a specific activity of 108.6 U/mg, with a molecular weight of 28.6 kDa, and was composed of 260 amino acid residues. AlyDS44 is a bifunctional alginate lyase, active on both polyguluronate and polymannuronate, though it preferentially degrades polyguluronate. The optimal pH of this enzyme is 8.5 and the optimal temperature is 45 °C. It is a salt-tolerant alginate lyase with an optimal activity at 0.6 M NaCl. Metal ions Mn2+, Co2+, and Fe2+ increased the alginate degrading activity, but it was inhibited in the presence of Zn2+ and Cu2+. The highly conserved regions of its amino acid sequences indicated that AlyDS44 belongs to the polysaccharide lyase family 7. The main breakdown products of the enzyme on alginate were disaccharides, trisaccharides, and tetrasaccharides, which demonstrated that this enzyme acted as an endo-type alginate lyase. AlyDS44 is a novel enzyme, with the potential for efficient production of alginate oligosaccharides with low degrees of polymerization.
Supplementation of fertiliser with the biostimulant molasses enhances hemp (Cannabis sativa) seed functional food antioxidant capacity by induction of stress responses
(Elsevier B V, 2024-08-01) Wise K; Williams LB; Selby-Pham S; Wright PFA; Simovich T; Gill H; Gupta A; Puri M; Selby-Pham J
Hemp is low-tetrahydrocannabinol (THC) Cannabis sativa that is cultivated globally for food and fibre. Hemp seeds and seed-oil are popularly consumed individually or utilised within cooking, due to their flavour, nutrition, and functional food benefits. The functional food properties of hemp seed include its beneficial fatty acid (FA) profile and antioxidant activity, which are associated with reduced inflammation that benefits chronic conditions including cardiovascular disease and cancer. Maximising these functional food properties can offer extensive health benefits. Biostimulants – such as molasses – are fertiliser additives that are associated with improvements to plant growth, yield, and biochemical composition; however, they have been scarcely applied to hemp. Furthermore, due to the complex composition of biostimulants, understanding their modes of action poses a significant challenge, impeding the optimization of their usage and the realization of associated benefits. Accordingly, this study aimed to characterise the impact of molasses on the functional food properties of hemp seeds and explore biostimulant-induced biochemical changes as indicators of modes of effect. Although molasses treatment did not significantly alter the FA profile (p = 0.960 for ω-6:ω-3 FA ratio), the antioxidant capacity (as measured by ABTS) was significantly increased (3.8-fold increase, p = 0.008). Metabolite profiling and statistical modelling indicated that this change was likely associated with increases in several lipophilic antioxidant metabolites, including hydroxycinnamic acid amides and cannabisins. Comparison of root, leaf, seed, and sugar leaf tissue changes in phytohormones and metabolites indicated that the growth of hemp with molasses was predominantly associated with the induction of the plant's endogenous stress response within the roots. These results indicate that the biostimulant molasses is a beneficial fertiliser additive for enhancing the functional food (antioxidant) potential of hemp seeds, thereby improving the health-benefits imparted to consumers.