Effect of culture conditions at lab‐scale on metabolite composition and antibacterial and antibiofilm activities of Dunaliella tertiolecta

Dunaliella tertiolecta RCC6 was cultivated indoors in glass bubble column photobioreactors operated under batch and semi‐continuous regimens and using two different conditions of light and temperature. Biomass was harvested by centrifugation, frozen, and then lyophilized. The soluble material was obtained by sequential extraction of the lyophilized biomass with solvents with a gradient of polarity (hexane, ethyl acetate, and methanol) and its metabolic composition was investigated through nuclear magnetic resonance (NMR) spectroscopy. The effect of light on chlorophyll biosynthesis was clearly shown through the relative intensities of the 1H NMR signals due to pheophytins. The highest signal intensity was observed for the biomasses obtained at lower light intensity, resulting in a lower light availability per cell. Under high temperature and light conditions, the 1H NMR spectra of the hexane extracts showed an incipient accumulation of triacylglycerols. In these conditions and under semi‐continuous regimen, an enhancement of β‐carotene and sterols production was observed. The antibacterial and antibiofilm activities of the extracts were also tested. Antibacterial activity was not detected, regardless of culture conditions. In contrast, the minimal biofilm inhibitory concentrations (MBICs) against Escherichia coli for the hexane extract obtained under semi‐continuous regimen using high temperature and irradiance conditions was promising.

[1]  M. Calijuri,et al.  Bioproducts from microalgae biomass: Technology, sustainability, challenges and opportunities. , 2022, Chemosphere.

[2]  K. Agrawal,et al.  Microalgae Dunaliella as biofuel feedstock and β-carotene production: An influential step towards environmental sustainability , 2022, Energy Conversion and Management: X.

[3]  N. Cordeiro,et al.  Microalgae as Sustainable Biofactories to Produce High-Value Lipids: Biodiversity, Exploitation, and Biotechnological Applications , 2021, Marine drugs.

[4]  Z. Dubinsky,et al.  Unconventional high-value products from microalgae: A review. , 2021, Bioresource technology.

[5]  C. Cárdenas,et al.  Cyanobacteria and Eukaryotic Microalgae as Emerging Sources of Antibacterial Peptides , 2020, Molecules.

[6]  Vaseeharan Baskaralingam,et al.  Identification and characterization of bioactive pigment carotenoids from shrimps and their biofilm inhibition , 2020 .

[7]  N. Moheimani,et al.  Microalgae: A potential sustainable commercial source of sterols , 2020 .

[8]  Porooshat Dadgostar,et al.  Antimicrobial Resistance: Implications and Costs , 2019, Infection and drug resistance.

[9]  S. Casal,et al.  Biorefinery of Dunaliella salina: Sustainable recovery of carotenoids, polar lipids and glycerol. , 2019, Bioresource technology.

[10]  I. Probert,et al.  NMR characterization and evaluation of antibacterial and antiobiofilm activity of organic extracts from stationary phase batch cultures of five marine microalgae (Dunaliella sp., D. salina, Chaetoceros calcitrans, C. gracilis and Tisochrysis lutea). , 2019, Phytochemistry.

[11]  I. Probert,et al.  Inhibition of Bacterial and Fungal Biofilm Formation by 675 Extracts from Microalgae and Cyanobacteria , 2019, Antibiotics.

[12]  G. Dönmez,et al.  Bioactive Compounds Produced by Dunaliella species, Antimicrobial Effects and Optimization of the Efficiency , 2019 .

[13]  C. González-López,et al.  NMR Metabolomics as an Effective Tool To Unravel the Effect of Light Intensity and Temperature on the Composition of the Marine Microalgae Isochrysis galbana. , 2019, Journal of agricultural and food chemistry.

[14]  I. Linossier,et al.  Anti-Bacterial Adhesion Activity of Tropical Microalgae Extracts , 2018, Molecules.

[15]  Y. Ghasemi,et al.  Antibacterial Potential of Chlorella vulgaris and Dunaliella salina Extracts Against Streptococcus mutans , 2018 .

[16]  A. Tempone,et al.  Ergosterol isolated from the basidiomycete Pleurotus salmoneostramineus affects Trypanosoma cruzi plasma membrane and mitochondria , 2017, Journal of Venomous Animals and Toxins including Tropical Diseases.

[17]  H. Coutinho,et al.  Cholesterol and ergosterol affect the activity of Staphylococcus aureus antibiotic efflux pumps. , 2017, Microbial pathogenesis.

[18]  Kenneth Wei Min Tan,et al.  Nitrogen-induced metabolic changes and molecular determinants of carbon allocation in Dunaliella tertiolecta , 2016, Scientific Reports.

[19]  M. Bhattacharjee.  PHARMACEUTICALLY VALUABLE BIOACTIVE COMPOUNDS OF ALGAE , 2016 .

[20]  W. Ruth,et al.  Trimethylated homoserine functions as the major compatible solute in the globally significant oceanic cyanobacterium Trichodesmium , 2016, Proceedings of the National Academy of Sciences.

[21]  Sanjoy Banerjee,et al.  Availability and Utilization of Pigments from Microalgae , 2016, Critical reviews in food science and nutrition.

[22]  B. Morga,et al.  Antimicrobial Compounds from Eukaryotic Microalgae against Human Pathogens and Diseases in Aquaculture , 2016, Marine drugs.

[23]  I. Linossier,et al.  Rapid identification of osmolytes in tropical microalgae and cyanobacteria by (1)H HR-MAS NMR spectroscopy. , 2016, Talanta.

[24]  M. Kumar,et al.  Metabolomics: an emerging frontier of systems biology in marine macrophytes , 2016 .

[25]  F. Esposito,et al.  Bioactivity Screening of Microalgae for Antioxidant, Anti-Inflammatory, Anticancer, Anti-Diabetes, and Antibacterial Activities , 2016, Front. Mar. Sci..

[26]  M. Domingues,et al.  Lipidomic Approaches towards Deciphering Glycolipids from Microalgae as a Reservoir of Bioactive Lipids , 2016, Marine drugs.

[27]  N. Abu-Ghannam,et al.  Antibacterial Derivatives of Marine Algae: An Overview of Pharmacological Mechanisms and Applications , 2016, Marine drugs.

[28]  Andrew N Lane,et al.  Applications of NMR spectroscopy to systems biochemistry. , 2016, Progress in nuclear magnetic resonance spectroscopy.

[29]  J. E. Olsen,et al.  Whole-Genome Sequence of Staphylococcus aureus S54F9 Isolated from a Chronic Disseminated Porcine Lung Abscess and Used in Human Infection Models , 2015, Genome Announcements.

[30]  Y. Chow,et al.  The role of micronutrients and strategies for optimized continual glycerol production from carbon dioxide by Dunaliella tertiolecta , 2015, Biotechnology and bioengineering.

[31]  Chunxiang Hu,et al.  Effect of light intensity on physiological changes, carbon allocation and neutral lipid accumulation in oleaginous microalgae. , 2015, Bioresource technology.

[32]  N. Thajuddin,et al.  Biofilm Inhibitory Effect of Spirulina platensis Extracts on Bacteria of Clinical Significance , 2015, Proceedings of the National Academy of Sciences, India Section B: Biological Sciences.

[33]  J. Costa,et al.  Biologically Active Metabolites Synthesized by Microalgae , 2015, BioMed research international.

[34]  M. Stanley,et al.  Unlocking nature’s treasure-chest: screening for oleaginous algae , 2015, Scientific Reports.

[35]  Wei Zhang,et al.  Advances in Microalgae-Derived Phytosterols for Functional Food and Pharmaceutical Applications , 2015, Marine drugs.

[36]  M. Morais-Braga,et al.  Enhancement of the antibiotic activity of aminoglycosides by alpha-tocopherol and other cholesterol derivates. , 2014, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[37]  David Lebeaux,et al.  Biofilm-Related Infections: Bridging the Gap between Clinical Management and Fundamental Aspects of Recalcitrance toward Antibiotics , 2014, Microbiology and Molecular Reviews.

[38]  L. Gigova,et al.  Biological activity of microalgae can be enhanced by manipulating the cultivation temperature and irradiance , 2014, Central European Journal of Biology.

[39]  R. Su,et al.  Antifouling potential of the marine microalga Dunaliella salina , 2014, World Journal of Microbiology and Biotechnology.

[40]  Guangli Yu,et al.  Total Synthesis and Structure-Activity Relationship of Glycoglycerolipids from Marine Organisms , 2014, Marine drugs.

[41]  U. Römling,et al.  Characterization of Biofilm Formation and the Role of BCR1 in Clinical Isolates of Candida parapsilosis , 2013, Eukaryotic Cell.

[42]  J. Schleucher,et al.  Two-dimensional ³¹P,¹H NMR spectroscopic profiling of phospholipids in cheese and fish. , 2013, Journal of agricultural and food chemistry.

[43]  Vishal Gupta,et al.  Central metabolic processes of marine macrophytic algae revealed from NMR based metabolome analysis , 2013 .

[44]  P. McGinn,et al.  New diacylglyceryltrimethylhomoserines from the marine microalga Nannochloropsis granulata and their nitric oxide inhibitory activity , 2013, Journal of Applied Phycology.

[45]  M. Francavilla,et al.  A mixture of phytosterols from Dunaliella tertiolecta affects proliferation of peripheral blood mononuclear cells and cytokine production in sheep. , 2012, Veterinary immunology and immunopathology.

[46]  V. Cuomo,et al.  Extraction, characterization and in vivo neuromodulatory activity of phytosterols from microalga Dunaliella tertiolecta. , 2012, Current medicinal chemistry.

[47]  R. Airs,et al.  Type I and Type II chlorophyll-a transformation products associated with algal senescence , 2011 .

[48]  S. Molin,et al.  The clinical impact of bacterial biofilms , 2011, International Journal of Oral Science.

[49]  Johanna Bernáldez-Sarabia,et al.  Evaluation of the antibacterial activity of different species of phytoplankton , 2010 .

[50]  R. Luque,et al.  Phytosterols from Dunaliella tertiolecta and Dunaliella salina: a potentially novel industrial application. , 2010, Bioresource technology.

[51]  E. Ibáñez,et al.  Screening for bioactive compounds from algae. , 2010, Journal of pharmaceutical and biomedical analysis.

[52]  K. Srinivasakumar,et al.  In vitro Studies on Bactericidal Activity and Sensitivity Pattern of Isolated Marine Microalgae against Selective Human Bacterial Pathogens , 2009 .

[53]  M. Shariati,et al.  Dunaliella biotechnology: methods and applications , 2009, Journal of applied microbiology.

[54]  E. Ibáñez,et al.  Antimicrobial activity of sub- and supercritical CO2 extracts of the green alga Dunaliella salina. , 2008, Journal of food protection.

[55]  E. Ibáñez,et al.  Dunaliella salina microalga pressurized liquid extracts as potential antimicrobials. , 2006, Journal of food protection.

[56]  A. Segre,et al.  Metabolic profile of lettuce leaves by high‐field NMR spectra , 2005, Magnetic resonance in chemistry : MRC.

[57]  J. van Staden,et al.  Screening microalgae for some potentially useful agricultural and pharmaceutical secondary metabolites , 2004, Journal of Applied Phycology.

[58]  J. Hannan,et al.  Identification of trehalose and glycine betaine as compatible solutes in the moderately halophilic sulfate reducing bacterium, Desulfovibrio halophilus , 1996 .

[59]  A. Kawashima,et al.  Antibiotic effect of linolenic acid fromChlorococcum strain HS-101 andDunaliella primolecta on methicillin-resistantStaphylococcus aureus , 1995, Journal of Applied Phycology.

[60]  Roberto Rizzo,et al.  Lipid extracts from different algal species:1H and13C-NMR spectroscopic studies as a new tool to screen differences in the composition of fatty acids, sterols and carotenoids , 1992, Journal of Applied Phycology.

[61]  Y. Shibano,et al.  Expression of human P-glycoprotein in yeast cells--effects of membrane component sterols on the activity of P-glycoprotein. , 1991, Agricultural and biological chemistry.

[62]  S. Kellam,et al.  Antibacterial activity from marine microalgae in laboratory culture , 1989 .

[63]  Y. Kanazawa,et al.  Nitrogen-14 nuclear quadrupole coupling constants effective for nuclear relaxation in molecules of trimethylalkylammonium type , 1983 .

[64]  Jeffrey L. C. Wright.  The occurrence of ergosterol and (22E,24R)-24-ethylcholesta-5,7,22-trien-3β-ol in the unicellular chlorophyte Dunaliellatertiolecta , 1979 .

[65]  A. Ben‐Amotz,et al.  The Role of Glycerol in the Osmotic Regulation of the Halophilic Alga Dunaliella parva. , 1973, Plant physiology.

[66]  R. Guillard,et al.  Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt, and Detonula confervacea (cleve) Gran. , 1962, Canadian journal of microbiology.

[67]  H. Spoehr,et al.  CHLORELLIN, AN ANTIBACTERIAL SUBSTANCE FROM CHLORELLA. , 1944, Science.

[68]  R. Pratt.  STUDIES ON CHLORELLA VULGARIS. V. SOME PROPERTIES OF THE GROWTH-INHIBITOR FORMED BY CHLORELLA CELLS , 1942 .

[69]  M. Borowitzka.  Biology of microalgae , 2018 .

[70]  S. Krishnakumar,et al.  EVALUATION OF BIOACTIVE METABOLITES FROM HALOPHILIC MICROALGAE DUNALIELLA SALINA BY GC - MS ANALYSIS , 2013 .

[71]  A. Pheophorbides,et al.  NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY OF CHLOROPHYLL , 2009 .

[72]  R. Hancock,et al.  Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances , 2008, Nature Protocols.

[73]  A. Amrane,et al.  Effect of culture conditions of , 1998 .

[74]  T. Kashimoto,et al.  Antibiotic substances produced by a marine green alga, Dunaliella primolecta , 1993 .

[75]  B. Lustigman.  Comparison of antibiotic production from four ecotypes of the marine alga,Dunaliella , 1988, Bulletin of environmental contamination and toxicology.