Structural elucidation of the EPS of slime producing Brevundimonas vesicularis sp isolated from a paper machine

The slime forming bacteria Brevundimonas vesicalaris sp. was isolated from a paper mill and its EPS was produced on laboratory scale. After production, the exopolysaccharide (EPS) was purified and analysed for its purity and homogeneity, HPSEC revealed one distinct population with a molecular mass of more than 2,000 kDa. The protein content was around 9 w/w%. The sample was analysed to determine its chemical structure. The EPS was found to consist of rhamnose, glucose, galacturonic acid and glucuronic acid. Due to the presence of uronic acids the molar ratio between the four sugars found varies from 3:5:2:4 by sugar composition analyses after methanolysis to 1:11:1 found by NMR. A repeating unit with a molecular mass of 678 Da was confirmed by MALDI-TOF mass spectrometry after mild acid treatment. C-13 and H-1 hetero- and homonuclear 2D NMR spectroscopy of the native and partial hydrolysed EPS revealed a -->4)-alpha-L-GlcpA-(1-->4)-alpha-D-GalpA-(1-->4)-beta-L-Rhap-(1-->4)-beta-D-Glcp(1--> repeating unit, no non-sugar substituents were present. (C) 2002 Elsevier Science Ltd. All rights reserved.