Mechanistic Insights into Dideoxygenation in Gentamicin Biosynthesis

Autor(es):Mostrar menos -

Li, Sicong ^[1] ; Bury, Priscila dos Santos ^[2] ; Huang, Fanglu ^[3] ; Guo, Junhong ^[1] ; Sun, Guo ^[1] ; Reva, Anna ^[3] ; Huang, Chuan ^[1] ; Jian, Xinyun ^[1] ; Li, Yuan ^[1] ; Zhou, Jiahai ^[4] ; Deng, Zixin ^[1] ; Leeper, Finian J. ^[5] ; Leadlay, Peter F. ^[3] ; Dias, Marcio V. B. ^{[2, 6]} ; Sun, Yuhui ^[1]

Afiliação do(s) autor(es)

^[1] Wuhan Univ, Key Lab Combinatorial Biosynth & Drug Discovery, Minist Educ, Wuhan 430071 - Peoples R China

^[2] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, BR-05508000 Sao Paulo - Brazil

^[3] Univ Cambridge, Dept Biochem, Cambridge CB2 1GA - England

^[4] Chinese Acad Sci, CAS Key Lab Quantitat Engn Biol, Shenzhen Inst Synthet Biol, Shenzhen Inst Adv Technol, Shenzhen 518055 - Peoples R China

^[5] Univ Cambridge, Dept Chem, Cambridge CB2 1EW - England

^[6] Univ Warwick, Dept Chem, Coventry CV4 7AL, W Midlands - England

• Tipo de documento: Artigo Científico
• Fonte: ACS CATALYSIS; v. 11, n. 19, p. 12274-12283, OCT 1 2021.
• Citações Web of Science: 0

Resumo

Gentamicin is an important aminoglycoside antibiotic used for treatment of infections caused by Gram-negative bacteria. Although most of the biosynthetic pathways of gentamicin have been elucidated, a remaining intriguing question is how the intermediates JI-20A and JI-20B undergo a dideoxygenation to form gentamicin C complex. Here we show that the dideoxygenation process starts with GenP-catalyzed phosphorylation of JI-20A and JI-20Ba. The phosphorylated products are successively modified by concerted actions of two PLP (pyridoxal 5'-phosphate)-dependent enzymes: elimination of water and then phosphate by GenB3 and double bond migration by GenB4. Each of these reactions liberates an imine which hydrolyses to a ketone or aldehyde and is then reaminated by GenB3 using an amino donor. Importantly, crystal structures of GenB3 and GenB4 have guided site-directed mutagenesis to reveal crucial residues for the enzymes' functions. We propose catalytic mechanisms for GenB3 and GenB4, which shed light on the already unrivalled catalytic versatility of PLP-dependent enzymes. (AU)

• Processo FAPESP: 14/07843-6 - Análise estrutural de enzimas chaves para biossíntese de gentamicina e sisomicina
• Beneficiário: Priscila dos Santos Bury
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 15/09188-8 - Biossíntese de antibióticos poliéteres e aminoglicosídeos: investigação estrutural de enzimas não usuais ou com aplicabilidade em biologia sintética
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 10/15971-3 - Caracterização estrutural de enzimas envolvidas em vias de biossíntese de antibióticos com interesse biotecnológico
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 18/00351-1 - Biologia Estrutural aplicada às enzimas envolvidas na biossíntese de produtos naturais: aplicações biotecnológicas e entendimento molecular de reações enzimáticas pouco usuais
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Regular