Assuntos
Infecções por Coronavirus/epidemiologia , Acessibilidade aos Serviços de Saúde , Pneumonia Viral/epidemiologia , Centros de Traumatologia/organização & administração , Betacoronavirus , COVID-19 , Protocolos Clínicos , Colorado/epidemiologia , Cuidados Críticos/organização & administração , Humanos , Salas Cirúrgicas/organização & administração , Pandemias , Equipamento de Proteção Individual , Admissão e Escalonamento de Pessoal , Guias de Prática Clínica como Assunto , SARS-CoV-2 , Capacidade de Resposta ante Emergências , TelemedicinaRESUMO
BACKGROUND: The basidiomycetous yeast Xanthophyllomyces dendrorhous has been described as a potential biofactory for terpenoid-derived compounds due to its ability to synthesize astaxanthin. Functional knowledge of the genes involved in terpenoid synthesis would create opportunities to enhance carotenoid production. A thiolase enzyme catalyzes the first step in terpenoid synthesis. RESULTS: Two potential thiolase-encoding genes were found in the yeast genome; bioinformatically, one was identified as an acetyl-CoA C-acetyltransferase (ERG10), and the other was identified as a 3-ketoacyl Co-A thiolase (POT1). Heterologous complementation assays in Saccharomyces cerevisiae showed that the ERG10 gene from X. dendrorhous could complement the lack of the endogenous ERG10 gene in S. cerevisiae, thereby allowing cellular growth and sterol synthesis. X. dendrorhous heterozygous mutants for each gene were created, and a homozygous POT1 mutant was also obtained. This mutant exhibited changes in pigment composition and higher ERG10 transcript levels than the wild type strain. CONCLUSIONS: The results support the notion that the ERG10 gene in X. dendrorhous is a functional acetyl-CoA C-acetyltransferase essential for the synthesis of mevalonate in yeast. The POT1 gene would encode a functional 3-ketoacyl Co-A thiolase that is non-essential for cell growth, but its mutation indirectly affects pigment production.
Assuntos
Acetil-CoA C-Aciltransferase/genética , Basidiomycota/enzimologia , Basidiomycota/genética , Carotenoides/biossíntese , Acetil-CoA C-Acetiltransferase/genética , Acetil-CoA C-Acetiltransferase/metabolismo , Acetil-CoA C-Aciltransferase/metabolismo , Sequência de Bases , Basidiomycota/metabolismo , Vias Biossintéticas , DNA Fúngico/genética , Genes Fúngicos , Engenharia Metabólica/métodos , Mutação , Reação em Cadeia da Polimerase , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Esteróis/biossíntese , Terpenos/metabolismo , Xantofilas/metabolismoRESUMO
BACKGROUND: Xanthophyllomyces dendrorhous is a basidiomycetous yeast that synthesizes astaxanthin, a carotenoid with great biotechnological impact. The ergosterol and carotenoid synthetic pathways derive from the mevalonate pathway and involve cytochrome P450 enzymes. Among these enzymes, the CYP51 family, which is involved in ergosterol biosynthesis, is one of the most remarkable that has C14-demethylase activity. RESULTS: In this study, the CYP51 gene from X. dendrorhous was isolated and its function was analyzed. The gene is composed of ten exons and encodes a predicted 550 amino acid polypeptide that exhibits conserved cytochrome P450 structural characteristics and shares significant identity with the sterol C14-demethylase from other fungi. The functionality of this gene was confirmed by heterologous complementation in S. cerevisiae. Furthermore, a CYP51 gene mutation in X. dendrorhous reduced sterol production by approximately 40% and enhanced total carotenoid production by approximately 90% compared to the wild-type strain after 48 and 120 h of culture, respectively. Additionally, the CYP51 gene mutation in X. dendrorhous increased HMGR (hydroxy-methylglutaryl-CoA reductase, involved in the mevalonate pathway) and crtR (cytochrome P450 reductase) transcript levels, which could be associated with reduced ergosterol production. CONCLUSIONS: These results suggest that the CYP51 gene identified in X. dendrorhous encodes a functional sterol C14-demethylase that is involved in ergosterol biosynthesis.