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A combined physiological and proteomic approach to reveal lactic-acid-induced alterations in Lactobacillus casei Zhang and its mutant with enhanced lactic acid tolerance

  • Genomics, transcriptomics, proteomics
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Abstract

Lactobacillus casei has traditionally been recognized as a probiotic and frequently used as an adjunct culture in fermented dairy products, where acid stress is an environmental condition commonly encountered. In the present study, we carried out a comparative physiological and proteomic study to investigate lactic-acid-induced alterations in Lactobacillus casei Zhang (WT) and its acid-resistant mutant. Analysis of the physiological data showed that the mutant exhibited 33.8% higher glucose phosphoenolpyruvate:sugar phosphotransferase system activity and lower glycolytic pH compared with the WT under acidic conditions. In addition, significant differences were detected in both cells during acid stress between intracellular physiological state, including intracellular pH, H+-ATPase activity, and intracellular ATP pool. Comparison of the proteomic data based on 2D-DIGE and i-TRAQ indicated that acid stress invoked a global change in both strains. The mutant protected the cells against acid damage by regulating the expression of key proteins involved in cellular metabolism, DNA replication, RNA synthesis, translation, and some chaperones. Proteome results were validated by Lactobacillus casei displaying higher intracellular aspartate and arginine levels, and the survival at pH 3.3 was improved 1.36- and 2.10-fold by the addition of 50-mM aspartate and arginine, respectively. To our knowledge, this is the first demonstration that aspartate may be involved in acid tolerance in Lactobacillus casei. Results presented here may help us understand acid resistance mechanisms and help formulate new strategies to enhance the industrial applications of this species.

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Acknowledgements

This project was financially supported by the Key Program of the National Natural Science Foundation of China (No. 20836003), the National Natural Science Foundation of China (No. 30900013), Major State Basic Research Development Program of China (973 Program, No. 2010CB535014), and the Self-determined Research Program of Jiangnan University(JUSRP 21009).

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, People’s Republic of China

    Chongde Wu, Juan Zhang, Guocheng Du & Jian Chen

  2. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, People’s Republic of China

    Wei Chen

  3. School of Food Science and Technology, Jiangnan University, Wuxi, 214122, People’s Republic of China

    Miao Wang

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  1. Chongde Wu
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  2. Juan Zhang
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  3. Wei Chen
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  4. Miao Wang
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Correspondence to Guocheng Du or Jian Chen.

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Chongde Wu and Juan Zhang contributed equally to this paper.

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Fig. S1

2D-gels for spot-picker. A total of 1200 μμg of protein was focused on 24 cm pH 4–7 IEF strips and stained with Coomassie blue R-250. Indicated protein spots were picked and subjected to in-gel digestion and mass spectrometry. (DOC 426 kb)

Fig. S2

Hierarchical clustering of proteomic data arranged by functional categories according to the COG classification. C, Energy production and conversion; D, Cell-cycle control, mitosis and meiosis; E, Amino acid transporter and metabolism; F, Nucleotide transport and metabolism; G, Carbohydrate transport and metabolism; H, Coenzyme transport and metabolism; I, Lipid transport and metabolism; J, Translation, ribosomal structure and biogenesis; K, Transcription; L, Replication recombination and repair; M, Cell wall/membrane biogenesis; O, Posttranslational modification, protein turnover, chaperones; P, Inorganic ion transport and metabolism; R, General function prediction only; UN, Function unknown and not in COG. (DOC 311 kb)

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Wu, C., Zhang, J., Chen, W. et al. A combined physiological and proteomic approach to reveal lactic-acid-induced alterations in Lactobacillus casei Zhang and its mutant with enhanced lactic acid tolerance. Appl Microbiol Biotechnol 93, 707–722 (2012). https://doi.org/10.1007/s00253-011-3757-6

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