Structural characterization of neutral and acidic glycolipids from Thermus thermophilus HB8.

Yasuo Suda,Fumiaki Okazaki,Yushi Hasegawa,Koichi Fukase,Seiki Kuramitsu,Susumu Kokubo,Shoichi Kusumoto,Seiji Adachi,Richard H Barton

doi:10.1371/journal.pone.0035067

Yasuo Suda, Fumiaki Okazaki + Show 7 more

Open Access

https://doi.org/10.1371/journal.pone.0035067

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Journal: PloS one	Publication Date: Jul 16, 2012
Citations: 26	License type: CC BY 4.0

Affiliation: Osaka University, SPring-8, Kagoshima University

Abstract

The structural characterization of glycolipids from Thermus thermophilus HB8 was performed in this study. Two neutral and one acidic glycolipids were extracted and purified by the modified TLC-blotting method, after which their chemical structures were determined by chemical composition analysis, mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. The structure of one of the neutral glycolipids, NGL-A, was Galp(α1-6)GlcpNacyl(β1-2)Glcp(α1-)acyl2Gro, and the other, NGL-C, was Galf(β1-2)Galp(α1-6)GlcpNacyl(β1-2)Glcp(α1-)acyl2Gro. The structure of NGL-C was identical to that reported previously [Oshima, M. and Ariga, T. (1976) FEBS Lett. 64, 440]. Both neutral glycolipids shared a common structural unit found in the Thermus species. The acyl groups found in NGL-A and NGL-C, iso-type pentadecanoxy and heptadecanoxy fatty acid, were also the same as those found in this species. In contrast, the acidic glycolipid, AGL-B, possessed the structure of N-(((GlcpNAc(α1-)acyl2Gro)P-2)GroA)alkylamine. The alkyl group in AGL-B was an iso-type heptadecanyl, suggesting that the iso-type structure of the long alkyl chain is responsible for the thermal stability of the bacteria.

Highlights

Thermus thermophilus HB8 is an aerobic, rod-shaped, nonsporulating, Gram-negative eubacterium, which can grow at temperatures over 75uC [1]
We focused on cell surface glycolipids, which were extracted with chloroform/methanol at room temperature, and analyzed their structural characteristics by mass spectrometry (MS) and nuclear magnetic resonance (NMR)
It was suggested that the component B was an acidic glycolipid, and the other two were neutral ones

Summary

Introduction

Thermus thermophilus HB8 is an aerobic, rod-shaped, nonsporulating, Gram-negative eubacterium, which can grow at temperatures over 75uC [1]. The number of genes (ORF) is about 1/15 of humans, and the produced proteins are heat-stable and crystallized, so are suitable for detailed physicochemical analyses [2,3,4]. This bacterium is being used for the ‘‘Structural-Biological Whole Cell Project’’, which aims to understand all biological phenomena necessary for cell life based on the structural information of biomolecules [5,6]. The structures elucidated were compared to glycolipids from Deinococcus radiodurans [7,8,9], which is a Gram-negative and radiation-resistant bacterium possessing genomes similar to those of T. thermophilus [10]

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Results

Conclusion