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Molecular & Cellular Proteomics 5:1105-1118, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

A Proteome Reference Map and Proteomic Analysis of Bifidobacterium longum NCC2705^*,S

Jing Yuan^,,¶, Li Zhu^,¶, Xiankai Liu^,¶, Ting Li, Ying Zhang, Tianyi Ying, Bin Wang^||, Junjun Wang, Hua Dong, Erling Feng, Qiang Li, Jie Wang^**, Hongxia Wang^**, Kaihua Wei^**, Xuemin Zhang^**, Cuifeng Huang, Peitang Huang, Liuyu Huang, Ming Zeng^||, and Hengliang Wang^,

From the Beijing Institute of Biotechnology, State Key Laboratory of Pathogen and Biosecurity, 100071 Beijing, China, College of Food Science and Engineering, Northwest Sci-Tech University of Agriculture and Forestry, 712100 Yangling, China, ^** National Center of Biomedical Analysis, 100850 Beijing, China, and ^|| National Institute for the Control of Pharmaceutical and Biological Products, 100050 Beijing, China

A comprehensive proteomic study was carried out to identify and characterize proteins expressed by Bifidobacterium longum NCC2705. A total of 708 spots representing 369 protein entries were identified by MALDI-TOF-MS and/or ESI-MS/MS. Isoelectric point values estimated by gel electrophoresis matched closely with their predicted ones, although some discrepancies exist suggesting that post-translational protein modifications might be common in B. longum. The identified proteins represent 21.4% of the predicted 1727 ORFs in the genome and correspond to 30% of the predicted proteome. Moreover 95 hypothetical proteins were experimentally identified. This is the first compilation of a proteomic reference map for the important probiotic organism B. longum NCC2705. The study aimed to define a number of cellular pathways related to important physiological processes at the proteomic level. Proteomic comparison of glucose- and fructose-grown cells revealed that fructose and glucose are catabolized via the same degradation pathway. Interestingly the sugar-binding protein specific to fructose (BL0033) and Frk showed higher levels of expression in cells grown on fructose than on glucose as determined by semiquantitative RT-PCR. BL0033 time course and concentration experiments showed that the induction time and fructose concentration correlates to increased expression of BL0033. At the same time, an ABC (ATP-binding cassette) transporter ATP-binding protein (BL0034) was slightly up-regulated in cells grown on fructose compared with glucose. All of the above results suggest that the uptake of fructose into the cell may be conducted by a specific transport system in which BL0033 might play an important role.

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