Elucidation of the biosynthetic pathway of korormicin in Pseudoalteromonas JO10 and heterologous production in E. coli

• Grant number: 17/09695-2
• Support Opportunities: Scholarships abroad - Research
• Start date: September 05, 2017
• End date: September 04, 2018
• Field of knowledge: Engineering - Chemical Engineering
• Principal Investigator: Adilson José da Silva
• Grantee: Adilson José da Silva
• Host Investigator: Mattheos Koffas
• Host Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
• Institution abroad: Rensselaers Polytechnic Institute, United States
• Associated research grant: 16/10636-8 - From the cell factory to the Biodiesel-Bioethanol integrated biorefinery: a systems approach applied to complex problems in micro and macroscales, AP.BIOEN.TEM

Abstract

The emergence of multi-resistant bacteria poses a serious health threat worldwide, and it has driven the search for new antimicrobial agents. In this context, some natural products from marine environments appear as promising antibiotic candidates, among which is korormicin. Although its mechanism of action has not yet been fully elucidated, it is already known that this compound is a potent and selective inhibitor of a Na(+)-pumping respiratory enzyme (Na(+)-NQR), which is responsible for the membrane Na+ gradient in some Gram-negative bacteria. Inhibition of NQR hinders bacterial respiration and some multi-drug efflux pumps, giving korormicin the ability to affect vital processes of some bacteria, and possibly to rescue existing antibiotic drugs to which bacteria have developed resistance. Genome sequencing of Pseudoalteromonas J010 led to the identification of a natural product biosynthesis gene cluster (BGC), possibly related to korormicin, containing an operon composed of 6 genes (krmABCDEF). In this project, we propose to study this BGC using CRISPR/Cas9 and dCas9 -based methods to confirm its role in the synthesis of korormicin. Next, the genes coding for enzymes in the pathway will be cloned in E. coli using the ePathBrick platform. By this approach, we expect to define the biosynthetic pathway of korormicin and also to heterologously produce this compound for the first time. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MAYNARD, ADAM; BUTLER, NICOLE L.; ITO, TAKESHI; DA SILVA, ADILSON JOSE; MURAI, MASATOSHI; CHEN, TSUTE; KOFFAS, MATTHEOS A. G.; MIYOSHI, HIDETO; BARQUERA, BLANCA. Antibiotic Korormicin A Kills Bacteria by Producing Reactive Oxygen Species. Journal of Bacteriology, v. 201, n. 11, JUN 2019. (17/09695-2)

DA SILVA, ADILSON JOSE; CUNHA, JOSIVAN DE SOUZA; HREHA, TERI; MICOCCI, KELLI CRISTINA; SELISTRE-DE-ARAUJO, HELOISA SOBREIRO; BARQUERA, BLANCA; KOFFAS, MATTHEOS A. G.. Metabolic engineering of E. coli for pyocyanin production. METABOLIC ENGINEERING, v. 64, p. 15-25, MAR 2021. (19/11437-7, 17/09695-2)