MOLECULAR AND PROTEOMIC ANALYSIS OF PSYCHROTOLERANT Brevundimonas sp. CI19

ALVARENGA A. E; SADAÑOSKI M. A.; ZAPATA P.D.; VILLEGAS L.; AMOROSO M. J.

Congreso; 50th Annual Meeting Argentine Society for Biochemistry and Molecular Biology; 2014

SAIB

Brevundimonas sp. Ci19 was isolated from seawater samples from sub-Antarctic marine ecosystems. In preview studies showed production of α-L-rhamnosidase with a broad range of properties that allows its application in several biotechnological processes. The aim of this work was to analyze the differential protein expression by Brevundimonas sp. Ci19 in the presence of rhamnose or glucose as only carbon source under cold-adapted conditions by two-dimensional polyacrylamide gel electrophoresis. Simultaneously, primers were designed for amplification of α-L-rhamnosidase sequence. For proteomic analysis, the bacteria were incubated in LBM medium containing glucose or rhamnose, during 48 h at 15°C and 200 rpm. The cells were recollected and broken using press French. Thesupernatants of the cell lysates were used as protein samples. Two-dimensional gel electrophoresis of the whole proteins pattern of Brevundimonas sp. Ci19 revealed protein differential expression by rhamnose and glucose presence. Sequencing by MALDI-TOF of the more relevant proteins suggested that different regulation schemes are involved in response to carbon source. In relation of molecular study, 542 bp were amplified with designed primers and the sequence presented 57% identity to the conserved domain pfam05592, belonging to the superfamily cl18469 of rhamnosidases.