González Alberto, Osorio Héctor, Romero Stephanie, Méndez Patricia, Sepúlveda Muriel, Laporte Daniel, Gutierrez-Cutiño Marlen, Santander Rocío, Castro-Nallar Eduardo, Moenne Alejandra
Laboratory of Marine Biotechnology, Faculty of Chemistry and Biology, University of Santiago of Chile, Santiago, Chile.
Laboratorio Multidisciplinario, Instituto de Ciencias Biomédicas, Universidad Autónoma de Chile, Talca, Chile.
Front Plant Sci. 2022 Sep 20;13:955601. doi: 10.3389/fpls.2022.955601. eCollection 2022.
To analyze the mechanisms involved in anthracene (ANT) degradation in the marine alga , total RNA was obtained from the alga cultivated without ANT and with 5 μM of ANT for 24 h, and transcriptomic analyses were performed. A transcriptome was assembled, transcripts differentially expressed were selected, and those overexpressed were identified. Overexpressed transcripts potentially involved in ANT degradation were: one aromatic ring dioxygenase, three 2-oxoglutarate Fe (II) dioxygenases (2-OGDOs), and three dienelactone hydrolases that may account for anthraquinone, phthalic anhydride, salicylic acid, and phthalic acid production (pathway 1). In addition, two flavin adenine dinucleotide (FAD)-dependent monooxygenases, four cytP450 monooxygenases, two epoxide hydrolase, one hydroxyphenylpyruvic acid dioxygenase (HPPDO), and two homogentisic acid dioxygenases (HGDOs) were identified that may also participate in ANT degradation (pathway 2). Moreover, an alkane monooxygenase (alkB), two alcohol dehydrogenases, and three aldehyde dehydrogenases were identified, which may participate in linear hydrocarbon degradation (pathway 3). Furthermore, the level of transcripts encoding some of mentioned enzymes were quantified by qRT-PCR are in the alga cultivated with 5 μM of ANT for 0-48 h, and those more increased were 2-OGDO, HGDO, and alkB monooxygenase. Thus, at least three pathways for ANT and linear hydrocarbons degradation may be existed in . In addition, ANT metabolites were analyzed by gas chromatography and mass spectrometry (GC-MS), allowing the identification of anthraquinone, phthalic anhydride, salicylic acid, and phthalic acid, thus validating the pathway 1.
为分析海洋藻类中蒽(ANT)降解所涉及的机制,从未添加ANT以及添加5 μM ANT培养24小时的藻类中提取总RNA,并进行转录组分析。组装了转录组,选择了差异表达的转录本,并鉴定了过表达的转录本。可能参与ANT降解的过表达转录本有:一种芳香环双加氧酶、三种2-氧代戊二酸铁(II)双加氧酶(2-OGDOs)和三种二烯内酯水解酶,它们可能参与蒽醌、邻苯二甲酸酐、水杨酸和邻苯二甲酸的生成(途径1)。此外,还鉴定出两种黄素腺嘌呤二核苷酸(FAD)依赖性单加氧酶、四种细胞色素P450单加氧酶、两种环氧化物水解酶、一种对羟基苯丙酮酸双加氧酶(HPPDO)和两种尿黑酸双加氧酶(HGDOs),它们也可能参与ANT降解(途径2)。此外,还鉴定出一种烷烃单加氧酶(alkB)、两种醇脱氢酶和三种醛脱氢酶,它们可能参与直链烃的降解(途径3)。此外,通过qRT-PCR对藻类中培养0 - 48小时添加5 μM ANT时上述一些酶的编码转录本水平进行了定量,其中增加较多的是2-OGDO、HGDO和alkB单加氧酶。因此,藻类中可能至少存在三种ANT和直链烃降解途径。此外,通过气相色谱和质谱(GC-MS)分析了ANT代谢产物,鉴定出了蒽醌、邻苯二甲酸酐、水杨酸和邻苯二甲酸,从而验证了途径1。