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Pavel Hrouzek
Laboratory of Algal Biotechnology, Institute of Microbiology of the Czech Academy of Sciences—Center Algatech, Trebon, Czech Republic

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Article
Published: 25 August 2021 in mSphere
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The regulation of the production of cyanopeptides beyond microcystin is essential to understand their ecological role in complex microbial communities, e.g., harmful cyanobacterial blooms. The role of chemical communication between the cyanobacterium and the epibionts within its phycosphere is at an initial stage of research, and little is understood about its specificity.

ACS Style

Subhasish Saha; Paul-Adrian Bulzu; Petra Urajová; Jan Mareš; Grzegorz Konert; João Câmara Manoel; Markéta Macho; Daniela Ewe; Pavel Hrouzek; Jiří Masojídek; Rohit Ghai; Kumar Saurav. Quorum-Sensing Signals from Epibiont Mediate the Induction of Novel Microviridins in the Mat-Forming Cyanobacterial Genus Nostoc. mSphere 2021, 6, e0056221 .

AMA Style

Subhasish Saha, Paul-Adrian Bulzu, Petra Urajová, Jan Mareš, Grzegorz Konert, João Câmara Manoel, Markéta Macho, Daniela Ewe, Pavel Hrouzek, Jiří Masojídek, Rohit Ghai, Kumar Saurav. Quorum-Sensing Signals from Epibiont Mediate the Induction of Novel Microviridins in the Mat-Forming Cyanobacterial Genus Nostoc. mSphere. 2021; 6 (4):e0056221.

Chicago/Turabian Style

Subhasish Saha; Paul-Adrian Bulzu; Petra Urajová; Jan Mareš; Grzegorz Konert; João Câmara Manoel; Markéta Macho; Daniela Ewe; Pavel Hrouzek; Jiří Masojídek; Rohit Ghai; Kumar Saurav. 2021. "Quorum-Sensing Signals from Epibiont Mediate the Induction of Novel Microviridins in the Mat-Forming Cyanobacterial Genus Nostoc." mSphere 6, no. 4: e0056221.

Article
Published: 11 August 2021 in Applied and Environmental Microbiology
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All living organisms depend on iron as an essential cofactor for indispensable enzymes. However, the sources of bioavailable iron are often limited.

ACS Style

Tomáš Galica; Nicola Borbone; Jan Mareš; Andreja Kust; Alessia Caso; Germana Esposito; Kumar Saurav; Jan Hájek; Klára Řeháková; Petra Urajová; Valeria Costantino; Pavel Hrouzek. Cyanochelins, an Overlooked Class of Widely Distributed Cyanobacterial Siderophores, Discovered by Silent Gene Cluster Awakening. Applied and Environmental Microbiology 2021, 87, 1 .

AMA Style

Tomáš Galica, Nicola Borbone, Jan Mareš, Andreja Kust, Alessia Caso, Germana Esposito, Kumar Saurav, Jan Hájek, Klára Řeháková, Petra Urajová, Valeria Costantino, Pavel Hrouzek. Cyanochelins, an Overlooked Class of Widely Distributed Cyanobacterial Siderophores, Discovered by Silent Gene Cluster Awakening. Applied and Environmental Microbiology. 2021; 87 (17):1.

Chicago/Turabian Style

Tomáš Galica; Nicola Borbone; Jan Mareš; Andreja Kust; Alessia Caso; Germana Esposito; Kumar Saurav; Jan Hájek; Klára Řeháková; Petra Urajová; Valeria Costantino; Pavel Hrouzek. 2021. "Cyanochelins, an Overlooked Class of Widely Distributed Cyanobacterial Siderophores, Discovered by Silent Gene Cluster Awakening." Applied and Environmental Microbiology 87, no. 17: 1.

Methods and protocol
Published: 19 June 2021 in Applied Microbiology and Biotechnology
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Microalgal contamination in algal culture is a serious problem hampering the cultivation process, which can result in considerable economic and time losses. With the field of microalgal biotechnology on the rise, development of new tools for monitoring the cultures is of high importance. Here we present a case study of the detection of fast-growing green algae Chlorella vulgaris (as contaminant) in a diatom Phaeodactylum tricornutum culture using various approaches. We prepared mixed cultures of C. vulgaris and P. tricornutum in different cell-to-cell ratios in the range from 1:103 to 1:107. We compared the sensitivity among microscopy, cultivation-based technique, PCR, and qPCR. The detection of C. vulgaris contamination using light microscopy failed in samples containing cell ratios <1:105. Our results confirmed PCR/qPCR to provide the most reliable and sensitive results, with detection sensitivity close to 75 cells/mL. The method was similarly sensitive in a pure C. vulgaris culture as well as in a mixed culture containing 107-times more P. tricornutum cells. A next-generation sequencing analysis revealed a positive discrimination of C. vulgaris during DNA extraction. The method of cultivation media exchange from sea water to fresh water, preferred by the Chlorella contaminant, demonstrated a presence of the contaminant with a sensitivity comparable to PCR approaches, albeit with a much longer detection time. The results suggest that a qPCR/PCR-based approach is the best choice for an early warning in the commercial culturing of microalgae. This method can be conveniently complemented with the substitution-cultivation method to test the proliferating potential of the contaminant. • PCR-based protocol developed for detection of Chlorella cells. • Synergy of various approaches shows deeper insight into a presence of contaminants. • Positive/negative discrimination occurs during DNA extraction in mixed cultures. • Newly developed assays ready to use as in diagnostics of contamination.

ACS Style

Tomáš Grivalský; Antonín Střížek; Pavel Přibyl; Jaromír Lukavský; Radim Čegan; Roman Hobza; Pavel Hrouzek. Comparison of various approaches to detect algal culture contamination: a case study of Chlorella sp. contamination in a Phaeodactylum tricornutum culture. Applied Microbiology and Biotechnology 2021, 105, 5189 -5200.

AMA Style

Tomáš Grivalský, Antonín Střížek, Pavel Přibyl, Jaromír Lukavský, Radim Čegan, Roman Hobza, Pavel Hrouzek. Comparison of various approaches to detect algal culture contamination: a case study of Chlorella sp. contamination in a Phaeodactylum tricornutum culture. Applied Microbiology and Biotechnology. 2021; 105 (12):5189-5200.

Chicago/Turabian Style

Tomáš Grivalský; Antonín Střížek; Pavel Přibyl; Jaromír Lukavský; Radim Čegan; Roman Hobza; Pavel Hrouzek. 2021. "Comparison of various approaches to detect algal culture contamination: a case study of Chlorella sp. contamination in a Phaeodactylum tricornutum culture." Applied Microbiology and Biotechnology 105, no. 12: 5189-5200.

Special issue article
Published: 18 June 2021 in Physiologia Plantarum
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Cyanobacteria produce a variety of chemically diverse cyclic lipopeptides with potent antifungal activities. These cyclic lipopeptides have an amphipathic structure comprised of a polar peptide cycle and hydrophobic fatty acid side chain. Many have antibiotic activity against a range of human and plant fungal pathogens. This review article aims to summarize the present knowledge on the chemical diversity and cellular effects of cyanobacterial cyclic lipopeptides that display antifungal activity. Cyclic antifungal lipopeptides from cyanobacteria commonly fall into four structural classes; hassallidins, puwainaphycins, laxaphycins, and anabaenolysins. Many of these antifungal cyclic lipopeptides act through cholesterol and ergosterol-dependent disruption of membranes. In many cases, the cyclic lipopeptides also exert cytotoxicity in human cells, and a more extensive examination of their biological activity and structure–activity relationship is warranted. The hassallidin, puwainaphycin, laxaphycin, and anabaenolysin structural classes are unified through shared complex biosynthetic pathways that encode a variety of unusual lipoinitiation mechanisms and branched biosynthesis that promote their chemical diversity. However, the biosynthetic origins of some cyanobacterial cyclic lipopeptides and the mechanisms, which drive their structural diversification in general, remain poorly understood. The strong functional convergence of differently organized chemical structures suggests that the production of lipopeptide confers benefits for their producer. Whether these benefits originate from their antifungal activity or some other physiological function remains to be answered in the future. However, it is clear that cyanobacteria encode a wealth of new cyclic lipopeptides with novel biotechnological and therapeutic applications.

ACS Style

David P. Fewer; Jouni Jokela; Lassi Heinilä; Reidun Aesoy; Kaarina Sivonen; Tomáš Galica; Pavel Hrouzek; Lars Herfindal. Chemical diversity and cellular effects of antifungal cyclic lipopeptides from cyanobacteria. Physiologia Plantarum 2021, 1 .

AMA Style

David P. Fewer, Jouni Jokela, Lassi Heinilä, Reidun Aesoy, Kaarina Sivonen, Tomáš Galica, Pavel Hrouzek, Lars Herfindal. Chemical diversity and cellular effects of antifungal cyclic lipopeptides from cyanobacteria. Physiologia Plantarum. 2021; ():1.

Chicago/Turabian Style

David P. Fewer; Jouni Jokela; Lassi Heinilä; Reidun Aesoy; Kaarina Sivonen; Tomáš Galica; Pavel Hrouzek; Lars Herfindal. 2021. "Chemical diversity and cellular effects of antifungal cyclic lipopeptides from cyanobacteria." Physiologia Plantarum , no. : 1.

Journal article
Published: 04 May 2021 in Algal Research
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Pheophorbide a is the major photosensitizer present in algal biomass and its presence in food supplement was linked to human health issues in the past. Although several pheophorbide a quantification methods have been proposed including the United States Pharmacopeia-National Formulary protocol (USP-NFP), there are open methodological issues which make their results questionable. In the present study we have tested optimal extraction conditions for pheophorbide a from a disintegrated biomass of green algae Chlorella and Haematococcus, widely used as food supplement. Further we have compared the quantification of pheophorbide a in algal extracts using HPLC-HRMS with the spectrophotometric quantification after USP-NFP in Haematococcus biomass and oleoresin samples. We have demonstrated that USP-NFP provide considerably (1.7–2.4 times) higher pheophorbide contents compared to HPLC-HRMS quantification using a pheophorbide a analytical standard. This was mainly caused by the fact that USP-NFP integrates the concentration of all pheophorbide variants present in the samples. We have detected nine pheophorbide analogues in the original algal sample as well as in the final sample of USP-NFP, of which pheophorbide a represented only ~40%. However, this explains the values obtained by USP-NFP only partly. We also show that 60% acetone is the most suitable extraction solvent for pheophorbide a from various algal samples; however, we noted that such extracts might exhibit high activity of chlorophyllase generating pheophorbide a during the extraction process. This activity differed between studied Chlorella and Haematococcus biomass extracts. We have achieved the chlorophyllase inactivation by a short-term high-temperature treatment (90 °C, 5 min). Acceptable stability of pheophorbide a and only minor degradation of chlorophyll to pheophorbide a has been observed at these conditions. This study demonstrates the need for 1) an HPLC-DAD-based method employing multiple pheophorbide standards and 2) involvement of chlorophyllase inactivation protocol, to ensure proper quantification of pheophorbide content in algal biomass.

ACS Style

Dominika Činčárová; Jan Hájek; Martin Dobřichovský; Martin Lukeš; Pavel Hrouzek. Recommendations on the quantitative analysis of pheophorbides, photosensitizers present in algal biomass intended as food supplement. Algal Research 2021, 56, 102298 .

AMA Style

Dominika Činčárová, Jan Hájek, Martin Dobřichovský, Martin Lukeš, Pavel Hrouzek. Recommendations on the quantitative analysis of pheophorbides, photosensitizers present in algal biomass intended as food supplement. Algal Research. 2021; 56 ():102298.

Chicago/Turabian Style

Dominika Činčárová; Jan Hájek; Martin Dobřichovský; Martin Lukeš; Pavel Hrouzek. 2021. "Recommendations on the quantitative analysis of pheophorbides, photosensitizers present in algal biomass intended as food supplement." Algal Research 56, no. : 102298.

Preprint content
Published: 24 April 2021
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The regulation of oligopeptides production is essential in understanding their ecological role in complex microbial communities including harmful cyanobacterial blooms. The role of chemical communication between the cyanobacterium and the microbial community harboured as epibionts within its phycosphere is at an initial stage of research and little is understood about its specificity. Herein, we present insight into the role of a bacterial epibiont in regulating production of cyanobacterial oligopeptides microviridins, well-known elastase inhibitors with presumed anti-grazing effects, in an ecologically important cyanobacterial genus Nostoc. Heterologous expression and identification of specific signal molecules from the epibiont suggest the role of a quorum sensing-based interaction. Further, physiological experiments show an increase in microviridin production without affecting cyanobacterial growth and photosynthetic activity. Simultaneously, oligopeptides presenting a selective inhibition pattern provide support for their specific function in response to the presence of cohabitant epibionts. Thus, the chemical interaction revealed in our study provides an example of an interspecies signalling pathway monitoring the bacterial flora around the cyanobacterial filaments and induction of intrinsic species-specific metabolic responses. IMPORTANCE The regulation of cyanopeptide production beyond microcystin is essential to understand their ecological role in complex microbial communities, e.g. harmful cyanobacterial blooms. The role of chemical communication between the cyanobacterium and the epibionts within its phycosphere is at an initial stage of research and little is understood about its specificity. The frequency of cyanopeptide occurrence also demonstrates the need to understand the contribution of cyanobacterial peptides to the overall biological impact of cyanopeptides on aquatic organisms and vertebrates including humans. Our results shed light on the epibiont control of cyanopeptide production via quorum sensing mechanisms and we posit that such mechanisms may be widespread in natural cyanobacterial bloom community regulation.

ACS Style

Subhasish Saha; Paul-Adrian Bulzu; Petra Urajová; Jan Mareš; Grzegorz Konert; João Câmara Manoel; Markéta Macho; Daniela Ewe; Pavel Hrouzek; Jiří Masojídek; Rohit Ghai; Kumar Saurav. Quorum sensing signals from epibiont mediate the induction of bioactive peptides in mat-forming cyanobacteria Nostoc. 2021, 1 .

AMA Style

Subhasish Saha, Paul-Adrian Bulzu, Petra Urajová, Jan Mareš, Grzegorz Konert, João Câmara Manoel, Markéta Macho, Daniela Ewe, Pavel Hrouzek, Jiří Masojídek, Rohit Ghai, Kumar Saurav. Quorum sensing signals from epibiont mediate the induction of bioactive peptides in mat-forming cyanobacteria Nostoc. . 2021; ():1.

Chicago/Turabian Style

Subhasish Saha; Paul-Adrian Bulzu; Petra Urajová; Jan Mareš; Grzegorz Konert; João Câmara Manoel; Markéta Macho; Daniela Ewe; Pavel Hrouzek; Jiří Masojídek; Rohit Ghai; Kumar Saurav. 2021. "Quorum sensing signals from epibiont mediate the induction of bioactive peptides in mat-forming cyanobacteria Nostoc." , no. : 1.

Journal article
Published: 10 April 2021 in Biosensors
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Selenium (Se) is an element with many commercial applications as well as an essential micronutrient. Dietary Se has antioxidant properties and it is known to play a role in cancer prevention. However, the general population often suffers from Se deficiency. Green algae, such as Chlorella vulgaris, cultivated in Se-enriched environment may be used as a food supplement to provide adequate levels of Se. We used Raman microspectroscopy (RS) for fast, reliable, and non-destructive measurement of Se concentration in living algal cells. We employed inductively coupled plasma-mass spectrometry as a reference method to RS and we found a substantial correlation between the Raman signal intensity at 252 cm−1 and total Se concentration in the studied cells. We used RS to assess the uptake of Se by living and inactivated algae and demonstrated the necessity of active cellular transport for Se accumulation. Additionally, we observed the intracellular Se being transformed into an insoluble elemental form, which we further supported by the energy-dispersive X-ray spectroscopy imaging.

ACS Style

Martin Kizovský; Zdeněk Pilát; Mykola Mylenko; Pavel Hrouzek; Jan Kuta; Radim Skoupý; Vladislav Krzyžánek; Kamila Hrubanová; Olga Adamczyk; Jan Ježek; Silvie Bernatová; Tereza Klementová; Alžběta Gjevik; Martin Šiler; Ota Samek; Pavel Zemánek. Raman Microspectroscopic Analysis of Selenium Bioaccumulation by Green Alga Chlorella vulgaris. Biosensors 2021, 11, 115 .

AMA Style

Martin Kizovský, Zdeněk Pilát, Mykola Mylenko, Pavel Hrouzek, Jan Kuta, Radim Skoupý, Vladislav Krzyžánek, Kamila Hrubanová, Olga Adamczyk, Jan Ježek, Silvie Bernatová, Tereza Klementová, Alžběta Gjevik, Martin Šiler, Ota Samek, Pavel Zemánek. Raman Microspectroscopic Analysis of Selenium Bioaccumulation by Green Alga Chlorella vulgaris. Biosensors. 2021; 11 (4):115.

Chicago/Turabian Style

Martin Kizovský; Zdeněk Pilát; Mykola Mylenko; Pavel Hrouzek; Jan Kuta; Radim Skoupý; Vladislav Krzyžánek; Kamila Hrubanová; Olga Adamczyk; Jan Ježek; Silvie Bernatová; Tereza Klementová; Alžběta Gjevik; Martin Šiler; Ota Samek; Pavel Zemánek. 2021. "Raman Microspectroscopic Analysis of Selenium Bioaccumulation by Green Alga Chlorella vulgaris." Biosensors 11, no. 4: 115.

Journal article
Published: 15 December 2020 in Separations
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Global demand for natural pigments has increased in the past few years. Myxoxanthophyll, a glycosylated monocyclic carotenoid, is a pigment that occurs naturally in cyanobacteria but no scalable isolation process has been developed to obtain it from its natural source to date. In this study, myxoxanthophyll was isolated from unicellular cyanobacterium Synechocystis salina (S. salina) using high-performance countercurrent chromatography (HPCCC), where the lower phase of the biphasic solvent system composed of n-heptane–ethanol–water (2:4:4, v/v/v) was used as a mobile phase, whereas its upper phase was the stationary phase. For the HPCCC isolation, a multi-injection method was developed, and four consecutive sample injections (70 mg each) were performed, obtaining, in total, 20 mg of myxoxanthophyll, which was finally purified with high-performance liquid chromatography (HPLC). Overall, a final myxoxanthophyll yield of 15 mg (98% purity) was obtained. The target pigment showed a weak antioxidant and tyrosinase inhibitory effect, and exhibited immune-stimulating properties by activating human granulocytes. The results presented here form a basis for the large-scale production of myxoxanthophyll, and show the potential benefits of this pigment for human health.

ACS Style

Michaela Nováková; Tereza Fábryová; Doris Vokurková; Iva Dolečková; Jiří Kopecký; Pavel Hrouzek; Lenka Tůmová; José Cheel. Separation of the Glycosylated Carotenoid Myxoxanthophyll from Synechocystis Salina by HPCCC and Evaluation of Its Antioxidant, Tyrosinase Inhibitory and Immune-Stimulating Properties. Separations 2020, 7, 73 .

AMA Style

Michaela Nováková, Tereza Fábryová, Doris Vokurková, Iva Dolečková, Jiří Kopecký, Pavel Hrouzek, Lenka Tůmová, José Cheel. Separation of the Glycosylated Carotenoid Myxoxanthophyll from Synechocystis Salina by HPCCC and Evaluation of Its Antioxidant, Tyrosinase Inhibitory and Immune-Stimulating Properties. Separations. 2020; 7 (4):73.

Chicago/Turabian Style

Michaela Nováková; Tereza Fábryová; Doris Vokurková; Iva Dolečková; Jiří Kopecký; Pavel Hrouzek; Lenka Tůmová; José Cheel. 2020. "Separation of the Glycosylated Carotenoid Myxoxanthophyll from Synechocystis Salina by HPCCC and Evaluation of Its Antioxidant, Tyrosinase Inhibitory and Immune-Stimulating Properties." Separations 7, no. 4: 73.

Journal article
Published: 31 August 2020 in Toxins
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Man-made shallow fishponds in the Czech Republic have been facing high eutrophication since the 1950s. Anthropogenic eutrophication and feeding of fish have strongly affected the physicochemical properties of water and its aquatic community composition, leading to harmful algal bloom formation. In our current study, we characterized the phytoplankton community across three eutrophic ponds to assess the phytoplankton dynamics during the vegetation season. We microscopically identified and quantified 29 cyanobacterial taxa comprising non-toxigenic and toxigenic species. Further, a detailed cyanopeptides (CNPs) profiling was performed using molecular networking analysis of liquid chromatography-tandem mass spectrometry (LC-MS/MS) data coupled with a dereplication strategy. This MS networking approach, coupled with dereplication, on the online global natural product social networking (GNPS) web platform led us to putatively identify forty CNPs: fourteen anabaenopeptins, ten microcystins, five cyanopeptolins, six microginins, two cyanobactins, a dipeptide radiosumin, a cyclooctapeptide planktocyclin, and epidolastatin 12. We applied the binary logistic regression to estimate the CNPs producers by correlating the GNPS data with the species abundance. The usage of the GNPS web platform proved a valuable approach for the rapid and simultaneous detection of a large number of peptides and rapid risk assessments for harmful blooms.

ACS Style

Andreja Kust; Klára Řeháková; Jaroslav Vrba; Vincent Maicher; Jan Mareš; Pavel Hrouzek; Maria-Cecilia Chiriac; Zdeňka Benedová; Blanka Tesařová; Kumar Saurav. Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach. Toxins 2020, 12, 561 .

AMA Style

Andreja Kust, Klára Řeháková, Jaroslav Vrba, Vincent Maicher, Jan Mareš, Pavel Hrouzek, Maria-Cecilia Chiriac, Zdeňka Benedová, Blanka Tesařová, Kumar Saurav. Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach. Toxins. 2020; 12 (9):561.

Chicago/Turabian Style

Andreja Kust; Klára Řeháková; Jaroslav Vrba; Vincent Maicher; Jan Mareš; Pavel Hrouzek; Maria-Cecilia Chiriac; Zdeňka Benedová; Blanka Tesařová; Kumar Saurav. 2020. "Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach." Toxins 12, no. 9: 561.

Journal article
Published: 20 August 2020 in Molecules
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Heterocytous cyanobacteria are among the most prolific sources of bioactive secondary metabolites, including anabaenopeptins (APTs). A terrestrial filamentous Brasilonema sp. CT11 collected in Costa Rica bamboo forest as a black mat, was studied using a multidisciplinary approach: genome mining and HPLC-HRMS/MS coupled with bioinformatic analyses. Herein, we report the nearly complete genome consisting of 8.79 Mbp with a GC content of 42.4%. Moreover, we report on three novel tryptophan-containing APTs; anabaenopeptin 788 (1), anabaenopeptin 802 (2), and anabaenopeptin 816 (3). Furthermore, the structure of two homologues, i.e., anabaenopeptin 802 (2a) and anabaenopeptin 802 (2b), was determined by spectroscopic analysis (NMR and MS). Both compounds were shown to exert weak to moderate antiproliferative activity against HeLa cell lines. This study also provides the unique and diverse potential of biosynthetic gene clusters and an assessment of the predicted chemical space yet to be discovered from this genus.

ACS Style

Subhasish Saha; Germana Esposito; Petra Urajová; Jan Mareš; Daniela Ewe; Alessia Caso; Markéta Macho; Kateřina Delawská; Andreja Kust; Pavel Hrouzek; Josef Juráň; Valeria Costantino; Kumar Saurav. Discovery of Unusual Cyanobacterial Tryptophan-Containing Anabaenopeptins by MS/MS-Based Molecular Networking. Molecules 2020, 25, 3786 .

AMA Style

Subhasish Saha, Germana Esposito, Petra Urajová, Jan Mareš, Daniela Ewe, Alessia Caso, Markéta Macho, Kateřina Delawská, Andreja Kust, Pavel Hrouzek, Josef Juráň, Valeria Costantino, Kumar Saurav. Discovery of Unusual Cyanobacterial Tryptophan-Containing Anabaenopeptins by MS/MS-Based Molecular Networking. Molecules. 2020; 25 (17):3786.

Chicago/Turabian Style

Subhasish Saha; Germana Esposito; Petra Urajová; Jan Mareš; Daniela Ewe; Alessia Caso; Markéta Macho; Kateřina Delawská; Andreja Kust; Pavel Hrouzek; Josef Juráň; Valeria Costantino; Kumar Saurav. 2020. "Discovery of Unusual Cyanobacterial Tryptophan-Containing Anabaenopeptins by MS/MS-Based Molecular Networking." Molecules 25, no. 17: 3786.

Preprint
Published: 12 August 2020
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Man-made shallow fishponds in the Czech Republic have been facing a high eutrophication since 1950s. Anthropogenic eutrophication and feeding of fish have strongly affected the physico-chemical properties of water and its aquatic community composition leading to harmful algal bloom formation. In our current study, we have characterised the phytoplankton community across three hypertrophic ponds to assess the phytoplankton dynamics during the vegetation season. We microscopically identified and quantified 29 cyanobacterial taxa comprised of non-toxigenic and toxigenic species. Further, a detailed cyanopeptides (CNPs) profiling was performed using molecular networking analysis of liquid chromatography tandem mass spectrometry (LC–MS/MS) data coupled with dereplication strategy. This MS networking approach coupled with dereplication on online global natural product social networking (GNPS) web platform led us to putatively identify forty CNPs: fourteen anabaenopeptins, ten microcystins, five cyanopeptolins, six microginins, two cyanobactins, a dipeptide radiosumin, a cyclooctapeptide planktocyclin and epidolastatin12. We have applied the binary logistic regression to estimate the CNPs producer by correlating the GNPS data with the species abundance. Usage of The combination of molecular networking and dereplication on online global natural product social networking (GNPS) web platform has proved as a valuable approach for rapid and simultaneous detection of high number of peptides, and rapidly assessing the risk for harmful bloom.

ACS Style

Andreja Kust; Klára Řeháková; Jaroslav Vrba; Vincent Maicher; Jan Mareš; Pavel Hrouzek; Cecilia Chiriac; Zdeňka Benedová; Blanka Tesařová; Kumar Saurav. Insight Into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using a MS/MS Networking Approach. 2020, 1 .

AMA Style

Andreja Kust, Klára Řeháková, Jaroslav Vrba, Vincent Maicher, Jan Mareš, Pavel Hrouzek, Cecilia Chiriac, Zdeňka Benedová, Blanka Tesařová, Kumar Saurav. Insight Into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using a MS/MS Networking Approach. . 2020; ():1.

Chicago/Turabian Style

Andreja Kust; Klára Řeháková; Jaroslav Vrba; Vincent Maicher; Jan Mareš; Pavel Hrouzek; Cecilia Chiriac; Zdeňka Benedová; Blanka Tesařová; Kumar Saurav. 2020. "Insight Into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using a MS/MS Networking Approach." , no. : 1.

Preprint
Published: 23 July 2020
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Heterocytous cyanobacteria are among the most prolific source of bioactive secondary metabolites, including anabaenopeptins (APTs). A terrestrial filamentous Brasilonema sp. CT11 collected in Costa Rica bamboo forest, as black mat was studied using a multidisciplinary approach: genome mining and HPLC-HRMS/MS coupled with bionformatic analyses. Herein, we report the nearly complete genome consisting 8.79 Mbp with a GC content of 42.4%. Moreover, we report on three novel tryptophane-containing APTs; anabaenopeptin 788 (1), anabaenopeptin 802 (2) and anabaenopeptin 816 (3). Further, the structure of two homologues, i.e., anabaenopeptin 802 (2a) and anabaenopeptin 802 (2b) was determined by spectroscopic analysis (NMR and MS). Both compounds were shown to exert weak to moderate antiproliferative activity against HeLa cell lines. This study also provides the unique and diverse potential of biosynthetic gene clusters and an assessment of the predicted chemical space yet to be discovered from this genus.

ACS Style

Subhasish Saha; Germana Esposito; Petra Urajova; Jan Mareš; Daniela Ewe; Alessia Caso; Marketa Macho; Katerina Delawska; Andreja Kust; Pavel Hrouzek; Josef Juran; Valeria Costantino; Kumar Saurav. Discovery of Unusual Cyanobacterial Tryptophan-containing Anabaenopeptins by MS/MS Based Molecular Networking. 2020, 1 .

AMA Style

Subhasish Saha, Germana Esposito, Petra Urajova, Jan Mareš, Daniela Ewe, Alessia Caso, Marketa Macho, Katerina Delawska, Andreja Kust, Pavel Hrouzek, Josef Juran, Valeria Costantino, Kumar Saurav. Discovery of Unusual Cyanobacterial Tryptophan-containing Anabaenopeptins by MS/MS Based Molecular Networking. . 2020; ():1.

Chicago/Turabian Style

Subhasish Saha; Germana Esposito; Petra Urajova; Jan Mareš; Daniela Ewe; Alessia Caso; Marketa Macho; Katerina Delawska; Andreja Kust; Pavel Hrouzek; Josef Juran; Valeria Costantino; Kumar Saurav. 2020. "Discovery of Unusual Cyanobacterial Tryptophan-containing Anabaenopeptins by MS/MS Based Molecular Networking." , no. : 1.

Journal article
Published: 12 June 2020 in Harmful Algae
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Puwainaphycins (PUW) and minutissamides (MIN) are cyanobacterial lipopeptides found in various cyanobacterial species. The first possible target of human exposure to them is intestinal epithelium but effect of PUW/MIN on enterocytes is not known at all. Using differentiated Caco-2 cells, PUW F was found to be cytotoxic from 5 µM concentration based on lactate dehydrogenase release assay and total protein concentration. However, it is also able to induce production of interleukin 8 in non-cytotoxic concentrations 1 and 2.5 µM detected by ELISA. Effects of MIN A and C were similar but less pronounced compared to PUW F. On the other hand, MIN D was the least toxic compound with no significant pro-inflammatory effects. Surprisingly, pro-inflammatory activation of the cells by PUW F and MIN C resulted in an increase in tight junction (TJ) protein claudin 4 expression determined by western blot analysis and confirmed by confocal microscopy. Furthermore, decrease in expression of zonula occludens 3, another TJ protein, was observed after the exposure to PUW F. Taken together, these cytotoxic lipopeptides, especially PUW F, are to be studied more deeply due to their capability to activate and/or deregulate human enterocytes in low concentrations.

ACS Style

Ondřej Vašíček; Jan Hájek; Lucie Bláhová; Pavel Hrouzek; Pavel Babica; Lukáš Kubala; Lenka Šindlerová. Cyanobacterial lipopeptides puwainaphycins and minutissamides induce disruptive and pro-inflammatory processes in Caco-2 human intestinal barrier model. Harmful Algae 2020, 96, 101849 .

AMA Style

Ondřej Vašíček, Jan Hájek, Lucie Bláhová, Pavel Hrouzek, Pavel Babica, Lukáš Kubala, Lenka Šindlerová. Cyanobacterial lipopeptides puwainaphycins and minutissamides induce disruptive and pro-inflammatory processes in Caco-2 human intestinal barrier model. Harmful Algae. 2020; 96 ():101849.

Chicago/Turabian Style

Ondřej Vašíček; Jan Hájek; Lucie Bláhová; Pavel Hrouzek; Pavel Babica; Lukáš Kubala; Lenka Šindlerová. 2020. "Cyanobacterial lipopeptides puwainaphycins and minutissamides induce disruptive and pro-inflammatory processes in Caco-2 human intestinal barrier model." Harmful Algae 96, no. : 101849.

Journal article
Published: 16 January 2020 in Microorganisms
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Streptomyces are among the most promising genera in terms of production ability to biosynthesize a variety of bioactive secondary metabolites with pharmaceutical interest. Coinciding with the increase in genomic sequencing of these bacteria, mining of their genomes for biosynthetic gene clusters (BGCs) has become a routine component of natural product discovery. Herein, we describe the isolation and characterization of a Streptomyces tendae VITAKN with quorum sensing inhibitory (QSI) activity that was isolated from southern coastal part of India. The nearly complete genome consists of 8,621,231bp with a GC content of 72.2%. Sequence similarity networks of the BGCs detected from this strain against the Minimum Information about a Biosynthetic Gene Cluster (MIBiG) database and 3365 BGCs predicted by antiSMASH analysis of publicly available complete Streptomyces genomes were generated through the BiG-SCAPE-CORASON platform to evaluate its biosynthetic novelty. Crude extract analysis using high-performance liquid chromatography connected to high resolution tandem mass spectrometry (HPLC-HRMS/MS) and dereplication through the Global Natural Product Social Molecular Networking (GNPS) online workflow resulted in the identification of cyclic dipeptides (2, 5-diketopiperazines, DKPs) in the extract, which are known to possess QSI activity. Our results highlight the potential of genome mining coupled with LC-HRMS/MS and in silico tools (GNPS) as a valid approach for the discovery of novel QSI lead compounds. This study also provides the biosynthetic diversity of BGCs and an assessment of the predicted chemical space yet to be discovered.

ACS Style

Nabila Mohammed Ishaque; Ilia Burgsdorf; Jessie James Limlingan Malit; Subhasish Saha; Roberta Teta; Daniela Ewe; Krishnan Kannabiran; Pavel Hrouzek; Laura Steindler; Valeria Costantino; Kumar Saurav. Isolation, Genomic and Metabolomic Characterization of Streptomyces tendae VITAKN with Quorum Sensing Inhibitory Activity from Southern India. Microorganisms 2020, 8, 121 .

AMA Style

Nabila Mohammed Ishaque, Ilia Burgsdorf, Jessie James Limlingan Malit, Subhasish Saha, Roberta Teta, Daniela Ewe, Krishnan Kannabiran, Pavel Hrouzek, Laura Steindler, Valeria Costantino, Kumar Saurav. Isolation, Genomic and Metabolomic Characterization of Streptomyces tendae VITAKN with Quorum Sensing Inhibitory Activity from Southern India. Microorganisms. 2020; 8 (1):121.

Chicago/Turabian Style

Nabila Mohammed Ishaque; Ilia Burgsdorf; Jessie James Limlingan Malit; Subhasish Saha; Roberta Teta; Daniela Ewe; Krishnan Kannabiran; Pavel Hrouzek; Laura Steindler; Valeria Costantino; Kumar Saurav. 2020. "Isolation, Genomic and Metabolomic Characterization of Streptomyces tendae VITAKN with Quorum Sensing Inhibitory Activity from Southern India." Microorganisms 8, no. 1: 121.

Research article
Published: 14 January 2020 in Journal of Agricultural and Food Chemistry
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Microalgae accumulate bioavailable selenium containing amino acids (Se-AAs) and these are useful as a food supplement. While this accumulation has been studied in phototrophic algal cultures, little data exists for heterotrophic cultures. We have determined Se-AAs content, selenium/sulphur (Se/S) substitution rates and overall Se accumulation balance in photo- and heterotrophic Chlorella cultures. Laboratory trials revealed that heterotrophic cultures tolerate Se doses ~8-fold higher compared to phototrophic cultures, resulting in a ~2-3-fold higher Se-AAs content. In large-scale experiments both cultivation regimes provided comparable Se-AAs content. Outdoor phototrophic cultures accumulated up to 400 μg g-1 of total Se-AAs and exhibited a high level of Se/S substitution (5-10%) with 30-60% organic/total Se embedded in the biomass. A slightly higher content of Se-AAs and ratio of Se/S substitution was obtained for a heterotrophic culture in pilot-scale fermentors. The data presented here shows that heterotrophic Chlorella cultures provide an alternative for Se-enriched biomass production and provides information on Se-AAs content and speciation in different cultivation regimes.

ACS Style

Mykola Mylenko; Dai Long Vu; Jan Kuta; Karolína Ranglová; David Kubáč; Gergely Lakatos; Tomáš Grivalský; Martin Pablo Caporgno; João Artur Da Câmara Manoel; Jiří Kopecký; Jiří Masojídek; Pavel Hrouzek. Selenium Incorporation to Amino Acids in Chlorella Cultures Grown in Phototrophic and Heterotrophic Regimes. Journal of Agricultural and Food Chemistry 2020, 68, 1654 -1665.

AMA Style

Mykola Mylenko, Dai Long Vu, Jan Kuta, Karolína Ranglová, David Kubáč, Gergely Lakatos, Tomáš Grivalský, Martin Pablo Caporgno, João Artur Da Câmara Manoel, Jiří Kopecký, Jiří Masojídek, Pavel Hrouzek. Selenium Incorporation to Amino Acids in Chlorella Cultures Grown in Phototrophic and Heterotrophic Regimes. Journal of Agricultural and Food Chemistry. 2020; 68 (6):1654-1665.

Chicago/Turabian Style

Mykola Mylenko; Dai Long Vu; Jan Kuta; Karolína Ranglová; David Kubáč; Gergely Lakatos; Tomáš Grivalský; Martin Pablo Caporgno; João Artur Da Câmara Manoel; Jiří Kopecký; Jiří Masojídek; Pavel Hrouzek. 2020. "Selenium Incorporation to Amino Acids in Chlorella Cultures Grown in Phototrophic and Heterotrophic Regimes." Journal of Agricultural and Food Chemistry 68, no. 6: 1654-1665.

Preprint
Published: 31 December 2019
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Streptomyces, being one of the most promising genera due to its ability to synthesize a variety of bioactive secondary metabolites of pharmaceutical interest, here studied in relation to its genomic and metabolomic potential. Coinciding with the increase in sequenced data, mining of bacterial genomes for biosynthetic gene clusters (BGCs) has become a routine component of natural product discovery. Herein, we describe the isolation and characterization of a Streptomyces tendae VITAKN with quorum sensing inhibitory activity (QSI) that was isolated from southern coastal parts of India. The nearly complete genome consists of 8,621,231bp with a GC content of 72.2%. Utilizing the BiG-SCAPE-CORASON platform, a sequence similarity network predicted from this strain was evaluated through sequence similarity analysis with the MIBiG database and existing 3,365 BGCs predicted by antiSMASH analysis of publicly available complete Streptomyces genomes. Crude extract analyzed on LC-HRMS/MS and Global Natural Product Social Molecular Networking (GNPS) online workflow using dereplication resulted in the identification of cyclic dipeptides (2,5-diketopiperazines, DKPs) in the extract, which are known to possess QSI activity. Our results highlight the potential use of genomic mining coupled with LC-HRMS/MS and bionformatic tools (GNPS) as a potent approach for metabolome studies in discovering novel QSI lead compounds. This study also provides the biosynthetic diversity of these BGCs and an assessment of the predicted chemical space yet to be discovered.

ACS Style

Nabila Imthiyaz; Ilia Burgsdorf; Jessie James Limlingan Malit; Subhasish Saha; Roberta Teta; Daniela Ewe; Krishnan Kannabiran; Pavel Hrouzek; Laura Steindler; Valeria Costantino; Kumar Saurav. Isolation, Genomic and Metabolomic Characterization of Streptomyces tendae VITAKN with Quorum Sensing Inhibitory Activity from Southern India. 2019, 1 .

AMA Style

Nabila Imthiyaz, Ilia Burgsdorf, Jessie James Limlingan Malit, Subhasish Saha, Roberta Teta, Daniela Ewe, Krishnan Kannabiran, Pavel Hrouzek, Laura Steindler, Valeria Costantino, Kumar Saurav. Isolation, Genomic and Metabolomic Characterization of Streptomyces tendae VITAKN with Quorum Sensing Inhibitory Activity from Southern India. . 2019; ():1.

Chicago/Turabian Style

Nabila Imthiyaz; Ilia Burgsdorf; Jessie James Limlingan Malit; Subhasish Saha; Roberta Teta; Daniela Ewe; Krishnan Kannabiran; Pavel Hrouzek; Laura Steindler; Valeria Costantino; Kumar Saurav. 2019. "Isolation, Genomic and Metabolomic Characterization of Streptomyces tendae VITAKN with Quorum Sensing Inhibitory Activity from Southern India." , no. : 1.

Original article
Published: 05 August 2019 in Folia Microbiologica
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The rapid emergence of resistance in pathogenic bacteria together with a steep decline in economic incentives has rendered a new wave in the drug development by the pharmaceutical industry and researchers. Since cyanobacteria are recognized as wide producers of pharmaceutically important compounds, we investigated thirty-four cyanobacterial extracts prepared by solvents of different polarities for their antimicrobial potential. Almost all tested cyanobacterial strains exhibited some degree of antimicrobial bioactivity, with more general effect on fungal strains compared with bacteria. Surprisingly ~50% of cyanobacterial extracts exhibited specific activity against one or few bacterial indicator strains with Gram-positive bacteria being more affected. Extracts of two most promising strains were subjected to activity-guided fractionation and determination of the minimum inhibitory concentration (MIC) against selected bacterial and fungal isolates. Multiple fractions were responsible for their antimicrobial effect with MIC reaching low-micromolar concentrations and in some of them high level of specificity was recorded. Twenty-six bioactive fractions analyzed on LC-HRMS/MS and Global Natural Product Social Molecular Networking (GNPS) online workflow using dereplication resulted in identification of only forty-nine peptide spectrum matches (PSMs) with eleven unique metabolites spectrum matches (MSMs). Interestingly, only three fractions from Nostoc calcicola Lukešová 3/97 and four fractions from Desmonostoc sp. Cc2 showed the presence of unique MSMs suggesting the presence of unknown antimicrobial metabolites among majority of bioactive fractions from both the strains. Our results highlight potential for isolation and discovery of potential antimicrobial bioactive lead molecules from cyanobacterial extracts.

ACS Style

Kumar Saurav; Markéta Macho; Andreja Kust; Kateřina Delawská; Jan Hájek; Pavel Hrouzek. Antimicrobial activity and bioactive profiling of heterocytous cyanobacterial strains using MS/MS-based molecular networking. Folia Microbiologica 2019, 64, 645 -654.

AMA Style

Kumar Saurav, Markéta Macho, Andreja Kust, Kateřina Delawská, Jan Hájek, Pavel Hrouzek. Antimicrobial activity and bioactive profiling of heterocytous cyanobacterial strains using MS/MS-based molecular networking. Folia Microbiologica. 2019; 64 (5):645-654.

Chicago/Turabian Style

Kumar Saurav; Markéta Macho; Andreja Kust; Kateřina Delawská; Jan Hájek; Pavel Hrouzek. 2019. "Antimicrobial activity and bioactive profiling of heterocytous cyanobacterial strains using MS/MS-based molecular networking." Folia Microbiologica 64, no. 5: 645-654.

Journal article
Published: 16 October 2018 in Molecules
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Muscotoxins are cyanobacterial cyclic lipopeptides with potential applications in biomedicine and biotechnology. In this study, Desmonostoc muscorum CCALA125 strain extracts were enriched by polymeric resin treatment, and subjected to HPCCC affording three cyclic lipopeptides (1–3), which were further repurified by semi-preparative HPLC, affording 1, 2, and 3, with a purity of 86%, 92%, and 90%, respectively. The chemical identities of 2–3 were determined as muscotoxins A and B, respectively, by comparison with previously reported ESI-HRMS/MS data, whereas 1 was determined as a novel muscotoxin variant (muscotoxin C) using NMR and ESI-HRMS/MS data. Owing to the high yield (50 mg), compound 2 was broadly screened for its antimicrobial potential exhibiting a strong antifungal activity against Alternaria alternata, Monographella cucumerina, and Aspergillus fumigatus, with minimum inhibitory concentration (MIC) values of 0.58, 2.34, and 2.34 µg/mL; respectively, and weak antibacterial activity against Bacillus subtilis with a MIC value of 37.5 µg/mL. Compounds 1 and 3 were tested only against the plant pathogenic fungus Sclerotinia sclerotiorum due to their low yield, displaying a moderate antifungal activity. The developed chromatographic method proved to be an efficient tool for obtaining muscotoxins with potent antifungal properties.

ACS Style

José Cheel; Jan Hájek; Marek Kuzma; Kumar Saurav; Iva Smýkalová; Eliška Ondráčková; Petra Urajová; Dai Long Vu; Karine Faure; Jiří Kopecký; Pavel Hrouzek. Application of HPCCC Combined with Polymeric Resins and HPLC for the Separation of Cyclic Lipopeptides Muscotoxins A–C and Their Antimicrobial Activity. Molecules 2018, 23, 2653 .

AMA Style

José Cheel, Jan Hájek, Marek Kuzma, Kumar Saurav, Iva Smýkalová, Eliška Ondráčková, Petra Urajová, Dai Long Vu, Karine Faure, Jiří Kopecký, Pavel Hrouzek. Application of HPCCC Combined with Polymeric Resins and HPLC for the Separation of Cyclic Lipopeptides Muscotoxins A–C and Their Antimicrobial Activity. Molecules. 2018; 23 (10):2653.

Chicago/Turabian Style

José Cheel; Jan Hájek; Marek Kuzma; Kumar Saurav; Iva Smýkalová; Eliška Ondráčková; Petra Urajová; Dai Long Vu; Karine Faure; Jiří Kopecký; Pavel Hrouzek. 2018. "Application of HPCCC Combined with Polymeric Resins and HPLC for the Separation of Cyclic Lipopeptides Muscotoxins A–C and Their Antimicrobial Activity." Molecules 23, no. 10: 2653.

Research article
Published: 23 March 2018 in ACS Chemical Biology
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The pederin family includes a number of bioactive compounds isolated from symbiotic organisms of diverse evolutionary origin. Pederin is linked to beetle-induced dermatitis in humans and pederin family members possess potent antitumor activity caused by selective inhibition of the eukaryotic ribosome. Their biosynthesis is accomplished by a polyketide/non-ribosomal peptide synthetase machinery employing an unusual trans-acyltransferase mechanism. Here we report a novel pederin type compound, cusperin, from the free-living cyanobacterium Cuspidothrix issatschenkoi (earlier Aphanizomenon). The chemical structure of cusperin is similar to that of nosperin recently isolated from the lichen cyanobiont Nostoc sharing the tehrahydropyran moiety and major part of the linear backbone. However, the cusperin molecule is extended by a glycine residue and lacks one hydroxyl substituent. Pederins were previously thought to be exclusive to symbiotic relationships. However, C. issatschenkoi is a non-symbiotic planktonic organism and a frequent component of toxic water blooms. Cusperin is devoid of the cytotoxic activity reported for other pederin family members. Hence, our findings raise questions about the role of pederin analogues in cyanobacteria and broaden the knowledge of ecological distribution of this group of polyketides.

ACS Style

Andreja Kust; Jan Mareš; Jouni Jokela; Petra Urajová; Jan Hájek; Kumar Saurav; Kateřina Voráčová; David P. Fewer; Esa Haapaniemi; Perttu Permi; Klára Řeháková; Kaarina Sivonen; Pavel Hrouzek. Discovery of a Pederin Family Compound in a Nonsymbiotic Bloom-Forming Cyanobacterium. ACS Chemical Biology 2018, 13, 1123 -1129.

AMA Style

Andreja Kust, Jan Mareš, Jouni Jokela, Petra Urajová, Jan Hájek, Kumar Saurav, Kateřina Voráčová, David P. Fewer, Esa Haapaniemi, Perttu Permi, Klára Řeháková, Kaarina Sivonen, Pavel Hrouzek. Discovery of a Pederin Family Compound in a Nonsymbiotic Bloom-Forming Cyanobacterium. ACS Chemical Biology. 2018; 13 (5):1123-1129.

Chicago/Turabian Style

Andreja Kust; Jan Mareš; Jouni Jokela; Petra Urajová; Jan Hájek; Kumar Saurav; Kateřina Voráčová; David P. Fewer; Esa Haapaniemi; Perttu Permi; Klára Řeháková; Kaarina Sivonen; Pavel Hrouzek. 2018. "Discovery of a Pederin Family Compound in a Nonsymbiotic Bloom-Forming Cyanobacterium." ACS Chemical Biology 13, no. 5: 1123-1129.

Journal article
Published: 09 March 2018 in Journal of Chromatography B
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Quantification of selenated amino-acids currently relies on methods employing inductively coupled plasma mass spectrometry (ICP-MS). Although very accurate, these methods do not allow the simultaneous determination of standard amino-acids, hampering the comparison of the content of selenated versus non-selenated species such as methionine (Met) and selenomethionine (SeMet). This paper reports two approaches for the simultaneous quantification of Met and SeMet. In the first approach, standard enzymatic hydrolysis employing Protease XIV was applied for the preparation of samples. The second approach utilized methanesulfonic acid (MA) for the hydrolysis of samples, either in a reflux system or in a microwave oven, followed by derivatization with diethyl ethoxymethylenemalonate. The prepared samples were then analyzed by multiple reaction monitoring high performance liquid chromatography tandem mass spectrometry (MRM-HPLC-MS/MS). Both approaches provided platforms for the accurate determination of selenium/sulfur substitution rate in Met. Moreover the second approach also provided accurate simultaneous quantification of Met and SeMet with a low limit of detection, low limit of quantification and wide linearity range, comparable to the commonly used gas chromatography mass spectrometry (GC–MS) method or ICP-MS. The novel method was validated using certified reference material in conjunction with the GC-MS reference method.

ACS Style

Dai Long Vu; Karolína Ranglová; Jan Hájek; Pavel Hrouzek. Quantification of methionine and selenomethionine in biological samples using multiple reaction monitoring high performance liquid chromatography tandem mass spectrometry (MRM-HPLC-MS/MS). Journal of Chromatography B 2018, 1084, 36 -44.

AMA Style

Dai Long Vu, Karolína Ranglová, Jan Hájek, Pavel Hrouzek. Quantification of methionine and selenomethionine in biological samples using multiple reaction monitoring high performance liquid chromatography tandem mass spectrometry (MRM-HPLC-MS/MS). Journal of Chromatography B. 2018; 1084 ():36-44.

Chicago/Turabian Style

Dai Long Vu; Karolína Ranglová; Jan Hájek; Pavel Hrouzek. 2018. "Quantification of methionine and selenomethionine in biological samples using multiple reaction monitoring high performance liquid chromatography tandem mass spectrometry (MRM-HPLC-MS/MS)." Journal of Chromatography B 1084, no. : 36-44.