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Molecular & Cellular Proteomics 8:2321-2338, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Proteomics Analysis of Lactobacillus casei Zhang, a New Probiotic Bacterium Isolated from Traditional Home-made Koumiss in Inner Mongolia of China^*,

Rina Wu^,, Weiwei Wang^¶, Dongliang Yu^¶, Wenyi Zhang, Yan Li, Zhihong Sun, Junrui Wu, He Meng^||,** and Heping Zhang^,

From the The Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education, Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China,
||School of Agricultural and Biological, Shanghai Jiao Tong University, Shanghai 200240, China,
¶Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100080, China, and
College of Food Science, Shenyang Agricultural University, Shenyang 11061, China

Lactobacillus casei Zhang, isolated from traditional home-made koumiss in Inner Mongolia of China, was considered as a new probiotic bacterium by probiotic selection tests. We carried out a proteomics study to identify and characterize proteins expressed by L. casei Zhang in the exponential phase and stationary phase. Cytosolic proteins of the strain cultivated in de Man, Rogosa, and Sharpe broth were resolved by two-dimensional gel electrophoresis using pH 4–7 linear gradients. The number of protein spots quantified from the gels was 487 ± 21 (exponential phase) and 494 ± 13 (stationary phase) among which a total of 131 spots were identified by MALDI-TOF/MS and/or MALDI-TOF/TOF according to significant growth phase-related differences or high expression intensity proteins. Accompanied by the cluster of orthologous groups (COG), codon adaptation index (CAI), and GRAVY value analysis, the study provided a very first insight into the profile of protein expression as a reference map of L. casei. Forty-seven spots were also found in the study that showed statistically significant differences between exponential phase and stationary phase. Thirty-three of the spots increased at least 2.5-fold in the stationary phase in comparison with the exponential phase, including 19 protein spots (e.g. Hsp20, DnaK, GroEL, LuxS, pyruvate kinase, and GalU) whose intensity up-shifted above 3.0-fold. Transcriptional profiles were conducted to confirm several important differentially expressed proteins by using real time quantitative PCR. The analysis suggests that the differentially expressed proteins were mainly categorized as stress response proteins and key components of central and intermediary metabolism, indicating that these proteins might play a potential important role for the adaptation to the surroundings, especially the accumulation of lactic acid in the course of growth, and the physiological processes in bacteria cell.

To whom correspondence may be addressed. Tel.: 86-471-4319940; Fax: 86-471-4300122; E-mail: hepingdd{at}vip.sina.com.

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