Gluconacetobacter diazotrophicus levansucrase is involved in tolerance to NaCl, sucrose and desiccation, and in biofilm formation
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Gluconacetobacter diazotrophicus is a nitrogen-fixing bacterium and endophyte of sugarcane, which expresses levansucrase, a fructosyltransferase exoenzyme with sucrose hydrolytic and levan biosynthetic activities. As a result of their physical properties, the levan can provide protection against stress caused by abiotic or biotic factors and participate in the formation of biofilms. In this study, we investigated the construction and function of a levansucrase-defective mutant of G. diazotrophicus. The lsdA mutant showed a decreased tolerance (65.5%) to 50–150 mM NaCl and a decrease of 89% in 876 mM (30%) sucrose, a reduction (99%) in tolerance to desiccation after 18 h, and a decrease (36.9–58.5%) in the ability to form cell aggregates on abiotic surfaces. Complementation of the mutant with the complete lsdA gene leads to a recovery of the ability to grow on sucrose-containing medium and to form slimy colonies, the ability to form the cell aggregates on abiotic surfaces and the tolerance to NaCl. This report demonstrates the importance of levansucrase in environmental adaptation of G. diazotrophicus under high osmotic stress and in biofilm formation.
KeywordsLevansucrase Osmotic stress Biofilm Diazotroph
This work was supported with grants from Consejo Nacional de Ciencia y Tecnología—México (No. J31278-B) and from the Coordinación de la Investigación Científica—U.M.S.N.H. to J.J.V.A. M.L.V.H. and F.C.V. were recipients of PhD and MSc scholarships from the Consejo Nacional de Ciencia y Tecnología—México, respectively. The pBB1RMCS-3 plasmid was a kind gift from A. García de los Santos. We are grateful to J.S. Bayuelo-Jiménez for the use of the osmometer and sonicator and J. E. López-Meza for sequencing support.
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