Functional analysis of genetic modules encoded within plasmid pP32BP2 of psychrophile Psychrobacter sp. DAB_AL32B strain – plasmid role in biofilm formation, osmoprotection, cryoprotection, and glycine betaine, choline, and carnitine metabolism

Grant number: DI2013 012543
Funded by:

Project description:

Temperature, as one of the basic factors limiting life on Earth, has a considerable effect on life of many organisms. There is an increasing number of studies dedicated to bacteria adapted to live in low temperatures – psychrophiles. Although it is well known that these bacteria produce cold-active enzymes which may be used in many biotechnology applications, the general knowledge of these microorganisms’ biology and their environment adaptation mechanisms is still limited.

Recent studies have shown that genes encoding adaptive features are sometimes carried within mobile genetic elements (MGEs) such as plasmids. The detailed functional analyses of these elements may provide valuable insights into role of MGEs in adaptation and evolution of psychrophiles.

In this project, functional analyses of plasmid pP32BP2 of psychrophilic Psychrobacter sp. DAB_AL32B strain isolated from the Arctic, will be conducted. This plasmid carries not only plasmid backbone genetic modules (REP and PAR), but also three phenotypic modules, which may be responsible for host adaptation to extreme habitat. Proteins encoded within these modules are potentially involved in: (i) type 3 fimbriae formation (MRK) and (ii-iii) transport and metabolism of glycine betaine, choline, and carnitine (BCC and CAI). Special emphasis will be put on functional analyses of these phenotypic modules.

The impact of the MRK module on cells adhesion and biofilm formation will be analysed.  It will also be investigated whether the BCC module plays a role in osmo- and cryoprotection and whether the CAI module enables bacterial growth under anaerobic conditions. For both BCC and CAI modules their role in carnitine metabolism will be demonstrated.

The analyses will also include plasmid backbone modules, REP and PAR. Construction of cloning and expression vectors useful in genetic analyses of psychrophiles will be conducted.

This project will enrich the general knowledge of psychrophilic bacteria. These studies will also provide unique information on the role of MGEs in determining bacterial adaptation to extremely cold environments.