26abd7bfe21aa12403e30ee9a8101859 MarDru_a2020v19n9p477iENG.pdf f41a12ae79268a53d0d807b6627ce38217a24e4a MarDru_a2020v19n9p477iENG.pdf 70fef6ad0832e9f18944049688c5649ee772e667ec7b5300c2541dd7df3fb828 MarDru_a2020v19n9p477iENG.pdf Title: Insights into the Light Response of Skeletonema marinoi: Involvement of Ovothiol Subject: Diatoms are one of the most widespread groups of microalgae on Earth. They possess extraordinary metabolic capabilities, including a great ability to adapt to different light conditions. Recently, we have discovered that the diatom Skeletonema marinoi produces the natural antioxidant ovothiol B, until then identified only in clams. In this study, we investigated the light-dependent modulation of ovothiol biosynthesis in S. marinoi. Diatoms were exposed to different light conditions, ranging from prolonged darkness to low or high light, also differing in the velocity of intensity increase (sinusoidal versus square-wave distribution). The expression of the gene encoding the key ovothiol biosynthetic enzyme, ovoA, was upregulated by high sinusoidal light mimicking natural conditions. Under this situation higher levels of reactive oxygen species and nitric oxide as well as ovothiol and glutathione increase were detected. No ovoA modulation was observed under prolonged darkness nor low sinusoidal light. Unnatural conditions such as continuous square-wave light induced a very high oxidative stress leading to a drop in cell growth, without enhancing ovoA gene expression. Only one of the inducible forms of nitric oxide synthase, nos2, was upregulated by light with consequent production of NO under sinusoidal light and darkness conditions. Our data suggest that ovothiol biosynthesis is triggered by a combined light stress caused by natural distribution and increased photon flux density, with no influence from the daily light dose. These results open new perspectives for the biotechnological production of ovothiols, which are receiving a great interest for their biological activities in human model systems. Keywords: algae; antioxidant; diatoms; light; nitric oxide; ovothiol; oxidative stress Author: Alfonsina Milito, Ida Orefice, Arianna Smerilli, Immacolata Castellano, Alessandra Napolitano, Christophe Brunet and Anna Palumbo Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Sun Sep 20 13:15:30 2020 CEST ModDate: Thu Mar 3 13:33:24 2022 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2091743 bytes Optimized: yes PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JNLWDW+URWPalladioL-Ital Type 1 Custom yes yes no 681 0 XNYQKJ+URWPalladioL-Bold Type 1 Custom yes yes no 684 0 CGLOJB+URWPalladioL-BoldItal Type 1 Custom yes yes no 687 0 UJPFVT+URWPalladioL-Roma Type 1 Custom yes yes no 690 0 ZUAAPI+Rpxr Type 1 Builtin yes yes no 693 0 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Nitric Oxide (NO) Determination Item: Title: Reactive Oxygen Species (ROS) Determination Item: Title: Thiols Determination Item: Title: Data Analysis Item: Title: Nitric Oxide Synthase (Nos) Protein Sequence Analysis Item: Title: References Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: UJPFVT+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: UJPFVT+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: ZUAAPI+Rpxr FontSubset: true FirstChar: 11 LastChar: 124 FontDescriptor: FontName: ZUAAPI+Rpxr Flags: Symbolic FontBBox: -40, -283, 904, 728 FontFile: true Font: BaseFont: XUNDIL+EURM10 FontSubset: true FirstChar: 11 LastChar: 25 FontDescriptor: FontName: XUNDIL+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: JNLWDW+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 122 FontDescriptor: FontName: 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HQBQWV+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: VSHNAE+Rpcxr FontSubset: true FirstChar: 169 LastChar: 169 FontDescriptor: FontName: VSHNAE+Rpcxr Flags: Symbolic FontBBox: -212, -286, 1344, 973 FontFile: true Font: BaseFont: CGLOJB+URWPalladioL-BoldItal FontSubset: true FirstChar: 49 LastChar: 118 FontDescriptor: FontName: CGLOJB+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, -300, 1073, 935 FontFile: true EncodingDictionary: Differences: true XMP: application/pdf Alfonsina Milito, Ida Orefice, Arianna Smerilli, Immacolata Castellano, Alessandra Napolitano, Christophe Brunet and Anna Palumbo Diatoms are one of the most widespread groups of microalgae on Earth. They possess extraordinary metabolic capabilities, including a great ability to adapt to different light conditions. Recently, we have discovered that the diatom Skeletonema marinoi produces the natural antioxidant ovothiol B, until then identified only in clams. In this study, we investigated the light-dependent modulation of ovothiol biosynthesis in S. marinoi. Diatoms were exposed to different light conditions, ranging from prolonged darkness to low or high light, also differing in the velocity of intensity increase (sinusoidal versus square-wave distribution). The expression of the gene encoding the key ovothiol biosynthetic enzyme, ovoA, was upregulated by high sinusoidal light mimicking natural conditions. Under this situation higher levels of reactive oxygen species and nitric oxide as well as ovothiol and glutathione increase were detected. No ovoA modulation was observed under prolonged darkness nor low sinusoidal light. Unnatural conditions such as continuous square-wave light induced a very high oxidative stress leading to a drop in cell growth, without enhancing ovoA gene expression. Only one of the inducible forms of nitric oxide synthase, nos2, was upregulated by light with consequent production of NO under sinusoidal light and darkness conditions. Our data suggest that ovothiol biosynthesis is triggered by a combined light stress caused by natural distribution and increased photon flux density, with no influence from the daily light dose. These results open new perspectives for the biotechnological production of ovothiols, which are receiving a great interest for their biological activities in human model systems. 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