[SCAN] Amazing Extremophiles: the Journey from Basic Microbial Physiology to Applications

Scan Seminar

Title: Amazing Extremophiles: the Journey from Basic Microbial Physiology to Applications

Speaker: Helena Santos

From: Cell Physiology & NMR Lab, ITQB

Abstract

Extremophiles are microorganisms that can thrive in the most extreme environments and appeared to be an ideal source of robust enzymes and exotic metabolites. In particular, the discovery of hyperthermophiles was a great incentive to investigate the determinants of protein stability. Surprisingly, in addition to intrinsically highly stable proteins, hyperthermophiles possess also intracellular proteins that are not particularly stable, thus indicating alternative strategies for their stabilization in vivo. In fact, (hyper)thermophiles isolated from saline environments synthesize de novo low-molecular mass compounds that have not been found, or have been rarely encountered in mesophilic organisms leading to the view that these solutes could play a role in the stabilization of cell components at high temperature. At least in vitro, these solutes are highly efficient in the protection of enzymes and proteins from thermophilic as well as mesophilic origin against thermal inactivation and other stresses. Interestingly, the compatible solutes from hyperthermophiles are generally negatively charged, while mesophilic organisms accumulate primarily neutral or zwitterionic solutes. The question then arises whether those charged solutes were selected by organisms adapted to grow at high temperatures because they are more suitable to protect proteins and other cell components against thermal denaturation.
Our team discovered many compatible solutes, identified biosynthetic routes, characterized the relevant enzymes and in collaboration with X-ray crystallographers determined the structures of some of them. Moreover, we assessed the efficacy of the novel solutes to protect enzymes against heat inactivation and/or aggregation. In a few cases, deletion mutants were used to study the physiological role of these solutes. Recently, we used a yeast model of Parkinson´s disease overexpressing α synuclein, the major protein in intraneuronal inclusions found in the brains of patients. This cell model was further engineered to synthesize mannosylglycerate (MG). The results demonstrate the ability of this solute to inhibit the formation of α synuclein inclusions in vivo. MG acts as a chemical chaperone and stabilizes the protein by direct solute/protein interactions.

A few recent publications of our team:

MANNOSYLGLYCERATE AND DI-MYO-INOSITOL-PHOSPHATE HAVE INTERCHANGEABLE ROLES DURING ADAPTATION OF Pyrococcus furiosus TO HEAT STRESS
A. M. Esteves, S. K. Chandrayan, P. M. McTernan, N. Borges, M. W.W. Adams & H. Santos
Applied and Environmental Microbiology, published online 2 May 2014, doi:10.1128/AEM.00559-14.

X-RAY STRUCTURE OF A CDP-ALCOHOL PHOSPHATIDYLTRANSFERASE MEMBRANE ENZYME AND INSIGHTS INTO ITS CATALYTIC MECHANISM
P. Nogly, I. Gushchin, A. Remeeva, A. M. Esteves, A. Ishchenko, P. Ma, S. Grudinin, N. Borges, E. Round, I. Moraes, V. Borshchevskiy, H. Santos, V. Gordeliy & M. Archer
Nature Communications, in press.

MANNOSYLGLYCERATE – STRUCTURAL ANALYSIS OF BIOSYNTHESIS AND EVOLUTIONARY HISTORY
N. Borges, C. Jorge, L. G. Gonçalves, S. Gonçalves, P. M. Matias & H. Santos
Extremophiles, in press.

INHIBITION OF FORMATION OF α-SYNUCLEIN INCLUSIONS BY MANNOSYLGLYCERATE IN A YEAST MODEL OF PARKINSON´S DISEASE
C. Faria, C. D. Jorge, N. Borges, S. Tenreiro, T. F. Outeiro & H. Santos
Biochimica Biophysica Acta – General Subjects, 1830, 4065-4072 (2013).

MANNOSYLGLYCERATE STABILIZES STAPHYLOCOCCAL NUCLEASE WITH RESTRICTION OF SLOW BETA-SHEET MOTIONS
T. M. Pais, P. Lamosa, M. Matzapetakis, D. L. Turner & H. Santos
Protein Science, 21, 1126-1137 (2012).

THE THREE-DIMENSIONAL STRUCTURE OF MANNOSYL-3-PHOSPHOGLYCERATE PHOSPHATASE FROM Thermus thermophilus HB27: A NEW MEMBER OF THE HALOALKANOID ACID DEHALOGENASE SUPERFAMILY
S. Gonçalves, A. M. Esteves, H. Santos, N. Borges, & P. M. Matias
Biochemistry, 50, 9551-9567 (2011).

EVOLUTION OF THE BIOSYNTHESIS OF DI-MYO-INOSITOL PHOSPHATE, A MARKER OF ADAPTATION TO HOT MARINE ENVIRONMENTS
L. G. Gonçalves, N. Borges, F. Serra, P. L. Fernandes, H. Dopazo & H. Santos
Environmental Microbiology, 14, 691-701 (2012).

Thermococcus kodakaraensis MUTANTS DEFICIENT IN DI-MYO-INOSITOL PHOSPHATE USE ASPARTATE TO COPE WITH HEAT STRESS
N. Borges, R. Matsumi, T. Imanaka, H. Atomi & H. Santos
Journal of Bacteriology, 192, 191-197 (2010).

A UNIQUE BETA-1,2-MANNOSYLTRANSFERASE OF Thermotoga maritima THAT USES DI-MYO-INOSITOL PHOSPHATE AS MANNOSYL ACCEPTOR
M. V. Rodrigues, N. Borges, C. P. Almeida, P. Lamosa & H. Santos
Journal of Bacteriology, 191, 6105-6115 (2009).

A few Patent applications

• Identification of a bifunctional gene for mannosylglycerate synthesis and development of a high-scale-production heterologous system based on Saccharomyces cerevisiae
  M. S. da Costa, N. Empadinhas, L. Albuquerque & H. Santos
  Patente Internacional submetida em 17/10/2003 (processo nº 3398007Y). Requerente: BIOCANT.
• Glycerophosphoinositol as a stabilizer and/or preservative of biological materials
  H. Santos, P. Lamosa, N. D. Raven, L. Gafeira & M. V. Rodrigues
  Patente submetida em 24 de Março de 2005 ao European Patent Office (EP 05398004, EP 1705246). Requerente: IBET.
• Derivados sintéticos de manosilglicerato para a estabilização e/ou preservação de biomateriais
  P. Lamosa, T. Q. Faria, M. R. Ventura, C. D. Maycock & H. Santos
  Patente PAT 103442 P; 2006/02/24. Requerente: STAB VIDA, Lda.
• Hexose derivatives, preparation and uses thereof
  C. D. Maycock, R. Ventura, E. C. Lourenço, H. Santos & A. S. Miguel
  United States Provisional Patent, Application No. 61/953,392, filed March 14, 2014.