Name: लेखक स्पॉटलाइट: उद्योगों में सतत नवाचारों के लिए समुद्री शैवाल के बायोएक्टिव यौगिकों की खोज
Uploaded: 2023-11-21T14:20:00
Description: Watch this Scientific Journal Video about Exploring Seaweed's Bioactive Compounds for Sustainable Innovations in Industries at JoVE.com

इस काम को पुर्तगाली फाउंडेशन फॉर साइंस एंड टेक्नोलॉजी (FCT) द्वारा MARE-समुद्री और पर्यावरण विज्ञान केंद्र (UIDP/04292/2020 और UIDB/04292/2020), और एसोसिएट लेबोरेटरी ARNET (LA/P/0069/2020) को दी गई रणनीतिक परियोजनाओं के माध्यम से समर्थित किया गया था। FCT ने मार्टा वी. फ्रीटास (UI/BD/150957/2021) और तातियाना परेरा (2021) को दिए गए व्यक्तिगत डॉक्टरेट अनुदान को भी वित्त पोषित किया। 07791. बीडी)। इस काम को प्रोजेक्ट HP4A - हेल्दी पास्ता फॉर ऑल (सह-प्रचार संख्या 039952) द्वारा भी आर्थिक रूप से समर्थित किया गया था, जो ERDF द्वारा सह-वित्त पोषित - यूरोपीय क्षेत्रीय विकास कोष, पुर्तगाल 2020 कार्यक्रम के तहत, COMPETE 2020 - प्रतिस्पर्धात्मकता और अंतर्राष्ट्रीयकरण परिचालन कार्यक्रम के माध्यम से किया गया था।

1. बायोमास कटाई और तैयारी
<ol><li>कम ज्वार के दौरान जी ग्रैसिलिस के नमूनों की कटाई करें और सुखाने, प्रकाश और हवा के संपर्क से बचने के लिए उन्हें अंधेरे, ठंडा बक्से में प्रयोगशाला में जल्दी से ले जाएं।</li>
	<li>?...

Gracilaria gracilis का दोहन: सतत नवाचार के लिए बायोएक्टिव क्षमता

एक तरल माध्यम में रोगाणुरोधी गतिविधि परीक्षण एक तरल माध्यम में निलंबित सूक्ष्मजीवों के खिलाफ रोगाणुरोधी पदार्थों की प्रभावशीलता का मूल्यांकन करने के लिए उपयोग किया जाता है और आमतौर पर विकास को रोकन?...

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	<tbody>
		<tr>
			<td>Absolute Ethanol</td>
			<td>Aga, Portugal</td>
			<td>64-17-5</td>
			<td></td>
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		<tr>
			<td>Ammonium Chloride</td>
			<td>PanReac</td>
			<td>12125-02-9</td>
			<td></td>
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			<td>Amphotericin B</td>
			<td>Sigma-Aldrich</td>
			<td>1397-89-3</td>
			<td></td>
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		<tr>
			<td>Analytical scale balance</td>
			<td>Sartorius, TE124S</td>
			<td>22105307</td>
			<td></td>
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			<td>Bacillus subtilis subsp. spizizenii</td>
			<td>German Collection of Microorganisms and Cell Cultures (DSMZ)</td>
			<td>DSM 347</td>
			<td></td>
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		<tr>
			<td>Biotin</td>
			<td>Panreac AppliChem</td>
			<td>58-85-5</td>
			<td></td>
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		<tr>
			<td>Centrifuge</td>
			<td>Eppendorf, 5810R</td>
			<td>5811JH490481</td>
			<td></td>
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		<tr>
			<td>Chloramphenicol</td>
			<td>PanReac</td>
			<td>56-75-7</td>
			<td></td>
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		<tr>
			<td>CO2 Chamber</td>
			<td>Memmert</td>
			<td>N/A</td>
			<td></td>
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			<td>Cool White Fluorescent Lamps</td>
			<td>OSRAM Lumilux Skywhite</td>
			<td>N/A</td>
			<td></td>
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			<td>Densitometer McFarland</td>
			<td>Grant Instruments</td>
			<td>N/A</td>
			<td></td>
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		<tr>
			<td>DMEM medium</td>
			<td>Sigma-Aldrich</td>
			<td>D5796</td>
			<td></td>
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		<tr>
			<td>DMSO</td>
			<td>Sigma-Aldrich</td>
			<td>67-68-5</td>
			<td></td>
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			<td>DPPH</td>
			<td>Sigma, Steinheim, Germany</td>
			<td>1898-66-4</td>
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			<td>Escherichia coli (DSM 5922)</td>
			<td>German Collection of Microorganisms and Cell Cultures (DSMZ)</td>
			<td>DSM5922</td>
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		<tr>
			<td>Ethanol 96%</td>
			<td>AGA-Portugal</td>
			<td>64-17-5</td>
			<td></td>
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		<tr>
			<td>Ethylenediaminetetraacetic Acid Disodium Salt Dihydrate (Na2EDTA)</td>
			<td>J.T.Baker</td>
			<td>6381-92-6</td>
			<td></td>
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		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Sigma-Aldrich</td>
			<td>F7524</td>
			<td></td>
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		<tr>
			<td>Filter Paper (Whatman No.1)</td>
			<td>Whatman</td>
			<td>WHA1001320</td>
			<td></td>
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		<tr>
			<td>Flasks</td>
			<td>VWR International, Alcabideche, Portugal&#160;</td>
			<td>N/A</td>
			<td></td>
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		<tr>
			<td>Folin-Ciocalteu</td>
			<td>VWR Chemicals</td>
			<td>31360.264</td>
			<td></td>
		</tr>
		<tr>
			<td>Gallic Acid&#160;</td>
			<td>Merck</td>
			<td>149-91-7</td>
			<td></td>
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		<tr>
			<td>Germanium (IV) Oxide, 99.999%</td>
			<td>AlfaAesar</td>
			<td>1310-53-8</td>
			<td></td>
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		<tr>
			<td>HaCaT cells &#8211; 300493</td>
			<td>CLS-Cell Lines Services, Germany&#160;</td>
			<td>300493</td>
			<td></td>
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		<tr>
			<td>Hot Plate Magnetic Stirrer</td>
			<td>IKA, C-MAG HS7</td>
			<td>06.090564</td>
			<td></td>
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		<tr>
			<td>Iron Sulfate</td>
			<td>VWR Chemicals</td>
			<td>10124-49-9</td>
			<td></td>
		</tr>
		<tr>
			<td>Laminar flow hood</td>
			<td>TelStar, Portugal</td>
			<td>526013</td>
			<td></td>
		</tr>
		<tr>
			<td>LB Medium&#160;</td>
			<td>VWR Chemicals</td>
			<td>J106</td>
			<td></td>
		</tr>
		<tr>
			<td>Listonella anguillarum</td>
			<td>German Collection of Microorganisms and Cell Cultures (DSMZ)</td>
			<td>&#160;DSM 21597</td>
			<td></td>
		</tr>
		<tr>
			<td>Manganese Chloride</td>
			<td>VWR Chemicals</td>
			<td>7773.01.5</td>
			<td></td>
		</tr>
		<tr>
			<td>Micropipettes&#160;</td>
			<td>Eppendorf, Portugal</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplates</td>
			<td>VWR International, Alcabideche, Portugal&#160;</td>
			<td>10861-666</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplates</td>
			<td>Greiner</td>
			<td>738-0168</td>
			<td></td>
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		<tr>
			<td>Microplates (sterile)</td>
			<td>Fisher Scientific</td>
			<td>10022403</td>
			<td></td>
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			<td>Microplate reader</td>
			<td>&#160;Epoch Microplate Spectrophotometer, BioTek, Vermont, USA</td>
			<td>1611151E</td>
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			<td>MTT</td>
			<td>Sigma-Aldrich</td>
			<td>289-93-1</td>
			<td></td>
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		<tr>
			<td>Muller-Hinton Broth (MHB)</td>
			<td>VWR Chemicals</td>
			<td>90004-658</td>
			<td></td>
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		<tr>
			<td>Oven</td>
			<td>Binder, FD115</td>
			<td>12-04490</td>
			<td></td>
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		<tr>
			<td>Oven</td>
			<td>Binder, BD115</td>
			<td>04-62615</td>
			<td></td>
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		<tr>
			<td>Penicillin</td>
			<td>Sigma-Aldrich</td>
			<td>1406-05-9</td>
			<td></td>
		</tr>
		<tr>
			<td>pH meter Inolab</td>
			<td>&#160;VWR International, Alcabideche, Portugal&#160;</td>
			<td>15212099</td>
			<td></td>
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		<tr>
			<td>Pippete tips</td>
			<td>Eppendorf, Portugal</td>
			<td>5412307</td>
			<td></td>
		</tr>
		<tr>
			<td>Pyrex Bottles Media Storage</td>
			<td>&#160;VWR International, Alcabideche, Portugal&#160;</td>
			<td>16157-169</td>
			<td></td>
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		<tr>
			<td>Rotary Evaporator</td>
			<td>Heidolph, Laborota 4000</td>
			<td>80409287</td>
			<td></td>
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		<tr>
			<td>Rotavapor</td>
			<td>IKA HB10, VWR International, Alcabideche, Portugal</td>
			<td>07.524254</td>
			<td></td>
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		<tr>
			<td>Sodium Carbonate (Na2CO3)</td>
			<td>Chem-Lab</td>
			<td>497-19-8</td>
			<td></td>
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		<tr>
			<td>Sodium Chloride (NaCl)&#160;</td>
			<td>Normax Chem</td>
			<td>7647-14-5</td>
			<td></td>
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		<tr>
			<td>Sodium Phosphate Dibasic</td>
			<td>Riedel-de Ha&#235;n</td>
			<td>7558-79-4</td>
			<td></td>
		</tr>
		<tr>
			<td>SpectraMagic NX</td>
			<td>Konica Minolta, Japan</td>
			<td></td>
			<td>color data analysis software</td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Evolution 201, Thermo Scientific, Madison, WI, USA</td>
			<td>5A4T092004</td>
			<td></td>
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		<tr>
			<td>Streptomycin</td>
			<td>Sigma-Aldrich</td>
			<td>57-92-1</td>
			<td></td>
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		<tr>
			<td>Thiamine</td>
			<td>Panreac AppliChem</td>
			<td>59-43-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin-EDTA</td>
			<td>Sigma-Aldrich</td>
			<td>T4049</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptic Soy Agar (TSA)</td>
			<td>VWR Chemicals</td>
			<td>ICNA091010617</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptic Soy Broth (TSB)&#160;</td>
			<td>VWR Chemicals</td>
			<td>22091</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrapure water&#160;</td>
			<td>Advantage A10 Milli-Q lab, Merck, Darmstadt, Germany</td>
			<td>F5HA17360B</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum pump</td>
			<td>Buchi, Switzerland</td>
			<td>FIS05-402-103</td>
			<td></td>
		</tr>
		<tr>
			<td>Vitamin B12</td>
			<td>Merck</td>
			<td>68-19-9</td>
			<td></td>
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valorization of the red seaweed gracilaria gracilis through a biorefinery approach

मूल्यवान और मल्टीटार्गेट बायोएक्टिव अवयवों को प्राप्त करने के लिए प्रचुर मात्रा में फीडस्टॉक के रूप में समुद्री शैवाल में रुचि लगातार बढ़ रही है। इस काम में, हम ग्रैसिलेरिया ग्रैसिलिस की क्षमता का पता लगाते हैं, जो कॉस्मेटिक, फार्माकोलॉजिकल, भोजन और फ़ीड अनुप्रयोगों के लिए अगर और अन्य अवयवों के स्रोत के रूप में अपने व्यावसायिक हित के लिए दुनिया भर में खेती की जाने वाली एक खाद्य लाल समुद्री शैवाल है।
जी ग्रैसिलिस विकास की स्थिति को वनस्पति प्रसार और स्पोरुलेशन के माध्यम से अनुकूलित किया गया था, जबकि एक बड़े बायोमास स्टॉक को प्राप्त करने के लिए भौतिक रासायनिक स्थितियों में हेरफेर किया गया था। समुद्री शैवाल बायोमास पर इथेनॉल और पानी के साथ ग्रीन निष्कर्षण पद्धतियों का प्रदर्शन किया गया था। अर्क की बायोएक्टिव क्षमता का मूल्यांकन इन विट्रो परख के एक सेट के माध्यम से किया गया था, जो उनके साइटोटॉक्सिसिटी, एंटीऑक्सिडेंट और रोगाणुरोधी गुणों से संबंधित था। इसके अतिरिक्त, सूखे समुद्री शैवाल बायोमास को भोजन के पोषण मूल्य को बढ़ाने के लिए पास्ता योगों में शामिल किया गया था। ग्रैसिलिस से निकाले गए पिगमेंट को भी दही में एक प्राकृतिक रंग के रूप में शामिल किया गया है, और उनकी स्थिरता का मूल्यांकन किया गया था। दोनों उत्पादों को बाजार तक पहुंचने से पहले सबसे अच्छा अंतिम सूत्रीकरण प्राप्त करने के उद्देश्य से एक अर्ध-प्रशिक्षित संवेदी पैनल की सराहना के लिए प्रस्तुत किया गया था।
परिणाम जी ग्रैसिलिस की बहुमुखी प्रतिभा का समर्थन करते हैं चाहे इसे पूरे बायोमास, अर्क और / कई अनुकूलित प्रोटोकॉल को लागू करने के माध्यम से, यह काम खाद्य, कॉस्मेटिक और जलीय कृषि बाजारों को लाभ पहुंचाने की क्षमता वाले उत्पादों के विकास की अनुमति देता है, पर्यावरणीय स्थिरता और एक नीली परिपत्र अर्थव्यवस्था को बढ़ावा देता है।
इसके अलावा, और एक बायोरेफाइनरी दृष्टिकोण के अनुरूप, अवशिष्ट समुद्री शैवाल बायोमास का उपयोग पौधों के विकास के लिए बायोस्टिमुलेंट के रूप में किया जाएगा या पुर्तगाल के लीरिया के घोड़ी-पॉलिटेक्निक के इन-हाउस जलीय कृषि प्रणालियों के जल शोधन में उपयोग किए जाने के लिए कार्बन सामग्री में परिवर्तित किया जाएगा।

&#160;Seaweeds can be regarded as an interesting natural raw material to be profited by the pharmaceutical, food, feed, and environmental sectors. They biosynthesize a panoply of molecules, many not found in terrestrial organisms, with relevant biological properties1,2. However, seaweed-optimized cultivation protocols need to be implemented to ensure a large biomass stock.
Cultivation methods must always consider the nature of the seaweed thalli and overall morphology. Gracilaria gracilis is a clonal taxon, meaning the attachment organ produces multiple vegetative axes. Propagation by fragmentation (vegetative reproduction) is thus achieved, as each of these axes is fully able to adopt an independent life from the main thallus3. Clonal taxa can be successfully integrated with simple and fast one-step cultivation methodologies, as large amounts of biomass are obtained by splitting the thallus into small fragments that quickly regenerate and grow into new, genetically identical individuals. Both haplontic and diplontic thalli may be used in this process. Although the genus exhibits a complex haplo-diplontic isomorphic triphasic life cycle, sporulation is rarely necessary except when genetic renewal of the stocks is required to achieve improved crops. In this case, both tetraspores (haplontic spores formed by meiosis) and carpospores (diplontic spores formed by mitosis) give rise to macroscopic thalli that can then be grown and propagated by vegetative reproduction4. Growth cycles are dictated by environmental conditions and the physiological state of the individuals, among other biological factors such as the emergence of epiphytes and the adhesion of other organisms. Therefore, optimizing growing conditions is crucial to ensure high productivity and produce good quality biomass5.
The extraction of bioactive compounds from seaweed, including G. gracilis, can be achieved through various methods6,7. The choice of the extraction method depends on the specific compounds of interest, the target application, and the characteristics of the seaweed. In this study, we focused on solvent extraction, which involves using green solvents, such as water or ethanol, to dissolve and extract bioactive compounds from the seaweed biomass. The extraction can be carried out through maceration in a versatile and effective way and can be used for a wide range of compounds. It is a simple and widely used method involving soaking biomass in a solvent for an extended period, typically at room or slightly elevated temperatures. The solvent is stirred to enhance the extraction process. After the desired extraction time, the solvent is separated from the solid material by filtration or centrifugation.
Water is a commonly used solvent in food applications due to its safety, availability, and compatibility with a wide range of food products. Water extraction is suitable for polar compounds such as polysaccharides, peptides, and certain phenolics. However, it may not effectively extract non-polar compounds. Ethanol is also a widely used solvent in food applications and can be effective for extracting a variety of bioactive molecules, including phenolic compounds, flavonoids, and certain pigments. Ethanol is generally recognized as safe for use in food and can be easily evaporated, leaving behind the extracted compounds. It is worth noting that the choice of extraction method should consider factors such as efficiency, selectivity, cost-effectiveness, and environmental impact. The optimization of extraction parameters, such as solvent concentration, extraction time, temperature, and pressure, is crucial to achieving optimal yields of bioactive compounds from G. gracilis or other seaweeds.
Seaweeds have been found to exhibit antimicrobial activity against a wide range of microorganisms, including bacteria, fungi, and viruses8. This activity is attributed to bioactive components, including phenolics, polysaccharides, peptides, and fatty acids. Several studies have demonstrated their efficacy against pathogens such as Escherichia coli, Staphylococcus aureus, Salmonella sp., and Pseudomonas aeruginosa, among others9. The antimicrobial activity of seaweeds is attributed to the presence of bioactive compounds that can interfere with microbial cell walls, membranes, enzymes, and signaling pathways10. These compounds may disrupt microbial growth, inhibit biofilm formation, and modulate immune responses.
Red seaweeds, also known as rhodophytes, are a group of algae that can exhibit antimicrobial activity against a variety of microorganisms. Within this group, G. gracilis contains various bioactive compounds that may contribute to its reported antimicrobial activity. While the specific molecules can vary, the common classes that have been reported in G. gracilis and may possess antimicrobial properties are polysaccharides, phenolics, terpenoids, and pigments11. However, it is important to note that the presence and amounts of these components can vary depending on factors such as the location of seaweed collection, seasonality, physiological condition of the thalli, and environmental conditions. Therefore, the specific class and concentration of antimicrobial compounds in G. gracilis may vary accordingly.
G. gracilis has also been found to hold antioxidant properties, containing various phenolic compounds, which have been shown to scavenge free radicals and reduce oxidative stress12. Antioxidants help to protect cells from damage caused by reactive oxygen species and have potential health benefits. Antioxidant capacity can be evaluated directly through different methods, including the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and, indirectly, through the quantification of total polyphenolic content (TPC)13.
Even though an ingredient is reported to have a prominent bioactivity, its cytotoxicity assessment is indispensable in evaluating natural and synthetic substances to be used in contact with living cells or tissues. There are several methods for measuring cytotoxicity, each one with advantages and limitations. Overall, they offer a range of options to evaluate the harmful effects of many substances on cells and, at the same time, to investigate the mechanisms of cell damage and death14.
In this work, we use the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, a colorimetric method introduced by Mosmann (1983)15. This method measures the reduction of tetrazolium salts to a purple formazan product by metabolically active cells. The higher the amount of formazan crystals, the higher the number of viable cells, thus providing an indirect measure of cytotoxicity14. Since in this work, G. gracilis water and ethanol extracts are intended to be incorporated into dermo-cosmetic formulations, the in vitro cytotoxicity evaluation is performed in a keratinocyte (HaCaT) cell line.
Concerning the food application, seaweeds are generally low in calories and nutritionally rich in dietary fibers, essential elements and amino acids, polysaccharides, polyunsaturated fatty acids, polyphenols, and vitamins2,16. G. gracilis is no exception, having an interesting nutritional value. Freitas et al. (2021)4 found that cultivated G. gracilis had higher levels of protein and vitamin C and maintained the level of total lipids compared to wild seaweed. This may represent an economic and environmental advantage, as nutritionally speaking, production is preferable to the exploitation of wild resources. In addition, consumers are increasingly concerned about the type of food they eat, so it is important to introduce new ingredients for food enrichment and use new resources to obtain extracts that can add value to a product and claim a &#34;clean label.&#34; Besides, the current market is very competitive, requiring the development of new products and innovative strategies to differentiate manufacturers from their competitors17.
The enrichment of products with poor nutritional value, such as pasta, with marine resources, including seaweed, is a strategy to introduce this resource as a new food and a market differentiation strategy through a product with distinct nutritional value. On the other hand, G. gracilis is a source of natural red pigments such as phycobiliproteins18, having high potential for applications in the food industry. This seaweed has shown high interest in several areas, and its application can be made using the whole seaweed, extracts and/or the remaining biomass. In this work, we demonstrate some examples of such applications.

लेखक स्पॉटलाइट: उद्योगों में सतत नवाचारों के लिए समुद्री शैवाल के बायोएक्टिव यौगिकों की खोज 

एक बायोरेफाइनरी दृष्टिकोण के माध्यम से लाल समुद्री शैवाल ग्रेसिलेरिया ग्रैसिलिस का वैलोराइजेशन

Our research is focused on the prospection of new bioactive compounds from seaweed that can be useful in different sectors such as food, cosmetic, and pharmaceutical industries. There is a growing awareness in society about the potential of marine organisms to deliver bioactive compounds. These compounds can be used to produce sustainable and safer functional cosmetics to improve the quality of food products and serve as source of new chemical structures for the development of new pharmaceutical drugs.One of the challenges of using seaweed in the development of new food products is to avoid using high amounts that could lead to unpleasant sensory properties in food. Using seaweed extracts is a way to get around this problem. With this protocol, we were able to develop pasta, wheat, add nutritional value by incorporating whole algae.Usually, the addition of these ingredients provides less common sensory properties. However, with the developed formulation, a consumer acceptance was still achieved.

रोगाणुरोधी गतिविधि
प्राप्त परिणामों की व्याख्या करते समय, यह ध्यान में रखा जाना चाहिए कि निषेध का प्रतिशत जितना अधिक होगा, उस विशिष्ट तनाव के विकास को रोकने म?...

Watch this Scientific Journal Video about Exploring Seaweed's Bioactive Compounds for Sustainable Innovations in Industries at JoVE.com

यहां, हम ग्रेसिलेरिया ग्रैसिलिस के एक एकीकृत वैलोराइजेशन के उद्देश्य से कई प्रोटोकॉल का वर्णन करते हैं: जंगली प्रजातियों की कटाई, घर में विकास और बायोएक्टिव अवयवों का निष्कर्षण। अर्क के एंटीऑक्सिडेंट, रोगाणुरोधी, और साइटोटोक्सिक प्रभावों का मूल्यांकन किया जाता है, साथ ही पूरे समुद्री शैवाल बायोमास और पिगमेंट से समृद्ध भोजन के पोषण और स्थिरता मूल्यांकन के साथ।

The interest in seaweeds as an abundant feedstock to obtain valuable and multitarget bioactive ingredients is continuously growing. In this work, we ...

हमारा शोध समुद्री शैवाल से नए बायोएक्टिव यौगिकों की संभावना पर केंद्रित है जो भोजन, कॉस्मेटिक और दवा उद्योगों जैसे विभिन्न क्षेत्रों में उपयोगी हो सकते हैं। जैव सक्रिय यौगिकों को वितरित करने के लिए समुद्री जीवों की क्षमता के बारे में समाज में जागरूकता बढ़ रही है। इन यौगिकों का उपयोग खाद्य उत्पादों की गुणवत्ता में सुधार के लिए टिकाऊ और सुरक्षित कार्यात्मक सौंदर्य प्रसाधनों का उत्पादन करने के लिए किया जा सकता है और नई दवा दवाओं के विकास के लिए नई रासायनिक संरचनाओं के स्रोत के रूप में काम किया जा सकता है।नए खाद्य उत्पादों के विकास में समुद्री शैवाल का उपयोग करने की चुनौतियों में से एक उच्च मात्रा का उपयोग करने से बचना है जिससे भोजन में अप्रिय संवेदी गुण हो सकते हैं। समुद्री शैवाल के अर्क का उपयोग करना इस समस्या को हल करने का एक तरीका है। इस प्रोटोकॉल के साथ, हम पास्ता, गेहूं विकसित करने में सक्षम थे, पूरे शैवाल को शामिल करके पोषण मूल्य जोड़ते हैं।आमतौर पर, इन अवयवों के अलावा कम सामान्य संवेदी गुण प्रदान करता है। हालांकि, विकसित सूत्रीकरण के साथ, एक उपभोक्ता स्वीकृति अभी भी हासिल की गई थी।

valorization-red-seaweed-gracilaria-gracilis-through-biorefinery

Research

JoVE Journal

Biology

Valorization of the Red Seaweed Gracilaria gracilis Through a Biorefinery Approach

The interest in seaweeds as an abundant feedstock to obtain valuable and multitarget bioactive ingredients is continuously growing. In this work, we explore the potential of Gracilaria gracilis, an edible red seaweed cultivated worldwide for its commercial interest as a source of agar and other ingredients for cosmetic, pharmacological, food, and feed applications.
G. gracilis growth conditions were optimized through vegetative propagation and sporulation while manipulating the physicochemical conditions to achieve a large biomass stock. Green extraction methodologies with ethanol and water were performed over the seaweed biomass. The bioactive potential of extracts was assessed through a set of in vitro assays concerning their cytotoxicity, antioxidant, and antimicrobial properties. Additionally, dried seaweed biomass was incorporated into pasta formulations to increase food&#39;s nutritional value. Pigments extracted from G. gracilis have also been incorporated into yogurt as a natural colorant, and their stability was evaluated. Both products were submitted to the appreciation of a semi-trained sensorial panel aiming to achieve the best final formulation before reaching the market.
Results support the versatility of G. gracilis whether it is applied as a whole biomass, extracts and/or pigments. Through implementing several optimized protocols, this work allows the development of products with the potential to profit the food, cosmetic, and aquaculture markets, promoting environmental sustainability and a blue circular economy.
Moreover, and in line with a biorefinery approach, the residual seaweed biomass will be used as biostimulant for plant growth or converted to carbon materials to be used in water purification of the in-house aquaculture systems of MARE-Polytechnic of Leiria, Portugal.

Here, we describe several protocols aiming at an integrated valorization of Gracilaria gracilis: wild species harvesting, in-house growth, and extraction of bioactive ingredients. The extracts&#39; antioxidant, antimicrobial, and cytotoxic effects are evaluated, along with the nutritional and stability assessment of food enriched with whole seaweed biomass and pigments.

Extraction of the Bioactive Ingredients from Gracilaria gracilis

Extraction of the Bioactive Ingredients from Gracilaria gracilis  

To begin, grow Gracilaria gracilis in 250 milliliters of flat bottom flasks in a climatic room. Once the biomass has been scaled up, dried, and ground, weigh the resulting dried biomass. Dissolve it in 100 milliliters of solvent and mix by stirring in a vessel protected from light for 30 minutes.Perform extractions with ethanol and water separately at room temperature. Filter the liquid extracts through filter paper to separate them from the remaining biomass. Centrifuge the extracts at 8, 000 G for 10 minutes, at room temperature.Freeze dry the aqueous extracts and use a rotary evaporator at 40 degrees Celsius to evaporate the ethanol extracts.

Gracilaria gracilis से Bioactive सामग्री का निष्कर्षण

Evaluation of the Antioxidant and Cytotoxic Effects of Gracilaria gracilis Extracts

Evaluation of the Antioxidant and Cytotoxic Effects of Gracilaria gracilis Extracts  

To begin, take the aqueous and ethanol extracts obtained from the dried Gracilaria gracilis. In a 96 well microplate, dispense two microliters of each sample and 198 microliters of DPPH dissolved in absolute ethanol. Then, using a microplate spectrophotometer, measure the absorbance at 517 nanometers.Take the human keratinocytes grown in 96 well plates overnight. Add two microliters of the DMSO dissolved extracts to 198 microliters of the medium in microplate wells. Remove the culture medium.Add 100 microliters of MTT to the cells and incubate in the dark for 30 minutes. To solubilize the intracellular formazan crystals, add 100 microliters of DMSO. Using a microplate reader, measure the absorbance at 570 nanometers.The DPPH test demonstrated that the temperature of 40 degrees Celsius showed higher inhibition values of oxidant activity compared to extracts obtained at room temperature or 70 degrees Celsius. No cytotoxic effects were observed on human keratinocytes, suggesting that at the maximum assay concentration, both aqueous and ethanol extracts are safe for cutaneous use.

Gracilaria gracilis अर्क के एंटीऑक्सीडेंट और साइटोटोक्सिक प्रभाव का मूल्यांकन

Pasta Production Using Gracilaria gracilis, Nutritional Evaluation, and Sensory Evaluation

Pasta Production Using Gracilaria gracilis, Nutritional Evaluation, and Sensory Evaluation  

To begin, combine predefined portions of rice flour, gracilaria gracilis, and chlorella vulgaris in the equipment, and add approximately 30%water to the mixture. After a few minutes of mixing, followed by extruding the pasta, using the equipment, cut it manually. To produce chifferi, dry the pasta at 68 degrees Celsius for 42 minutes, followed by an additional five hours and 30 minutes at 76 degrees Celsius, simulating an industrial process.Finally, vacuum seal the samples and store them in a dark place at room temperature. Take one gram sample in a glass crucible with a filter bottom and position it in the fiber analyzer. Add 150 milliliters of preheated 1.25%sulfuric acid, and two milliliters of anti-foaming agent to the column of each crucible.Remove the solvent and rinse the crucible three times with deionized water to prepare the sample for the basic hydrolysis. After washing the crucibles three times with 150 milliliters of acetone, carefully transfer those to an oven set at 150 degrees Celsius for one hour. Record the final weight.Transfer the crucibles to a muffle furnace, set at 500 degrees Celsius for three hours, and then record the final weight. To determine the fatty acids profile, add two milliliters of 2%sulfuric acid solution in methanol to a 50 milligram sample. Heat the mixture at 80 degrees Celsius for two hours.Then, cool the mixture to room temperature and add one milliliter of ultrapure water and two milliliters of N-heptane to each sample. After vortexing the mixture for one minute, centrifuge it for five minutes. Later, recover the upper N-heptane phase containing the fatty acid methyl esters and transfer it to gas chromatography vials.Analyze the samples in a gas chromatograph equipped with a TR-FAME capillary column, an auto sampler, and a flame ionization detector. Determine the fatty acid profile by comparing the resulting retention times with a standard. Incubate porcelain crucibles for three hours at 105 degrees Celsius.Allow them to cool in a desiccator, and then record their weights. After incubating 10 grams of the sample, as demonstrated previously, calculate its moisture content using the displayed equation. Then, transfer the crucibles with the dried samples to an incinerator heated to 525 degrees Celsius for four hours.After cooling the samples, record their weight. To assess consumer acceptance, use chifferi pasta samples cooked in distilled water for eight minutes. Conduct sensory tests in individual sensory booths located in a sensory analysis laboratory.Evaluate the visual appearance, color, texture, odor, sea taste, overall taste, overall evaluation, and purchase intent of the samples. The pasta received ratings of five and six on a hedonic scale of one to nine for sea taste and visual appearance, respectively. The pasta had a purchase intention score of five and an overall appreciation score of six.

Gracilaria gracilis, पोषण मूल्यांकन, और संवेदी मूल्यांकन का उपयोग कर पास्ता उत्पादन

Incorporation of Seaweed Extract in Yogurt and Color Stability Evaluation 

To begin, add 50 milliliters of sodium phosphate buffer to one gram of Gracilaria gracilis. After homogenizing the mixture, use a mortar and pestle to macerate the sample for 10 minutes. Transfer the solution to a tube and centrifuge it for 20 minutes at 12298g.After combining the supernatant, slowly add 65%ammonium sulfate, then pellet the precipitate by centrifugation and dissolve it in distilled water. Perform dialysis of the extract using a tubing membrane against water for 24 hours, followed by freeze drying. To incorporate the extract, mix it in yogurt at a concentration of 0.21%Store the samples in individual glass flasks at 4 degrees Celsius until analysis.To check the color stability of the extract in yogurt, calibrate the colorimeter using a white ceramic plate. Fill a cell with approximately 28 grams of the sample and analyze its color. The color stability of the yogurts with pigment was evaluated for 12 days at 4 degrees celsius.The incorporation of the extracts in yogurts showed good color attention with a color difference of 7.01 plus or minus 2.36 after 12 days of storage.