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http://dx.doi.org/10.4014/jmb.1508.08030

Enhanced Acid Tolerance in Bifidobacterium longum by Adaptive Evolution: Comparison of the Genes between the Acid-Resistant Variant and Wild-Type Strain  

Jiang, Yunyun (The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University)
Ren, Fazheng (The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University)
Liu, Songling (The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University)
Zhao, Liang (The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University)
Guo, Huiyuan (The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University)
Hou, Caiyun (The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University)
Publication Information
Journal of Microbiology and Biotechnology / v.26, no.3, 2016 , pp. 452-460 More about this Journal
Abstract
Acid stress can affect the viability of probiotics, especially Bifidobacterium. This study aimed to improve the acid tolerance of Bifidobacterium longum BBMN68 using adaptive evolution. The stress response, and genomic differences of the parental strain and the variant strain were compared by acid stress. The highest acid-resistant mutant strain (BBMN68m) was isolated from more than 100 asexual lines, which were adaptive to the acid stress for 10th, 20th, 30th, 40th, and 50th repeats, respectively. The variant strain showed a significant increase in acid tolerance under conditions of pH 2.5 for 2 h (from 7.92 to 4.44 log CFU/ml) compared with the wild-type strain (WT, from 7.87 to 0 log CFU/ml). The surface of the variant strain was also smoother. Comparative whole-genome analysis showed that the galactosyl transferase D gene (cpsD, bbmn68_1012), a key gene involved in exopolysaccharide (EPS) synthesis, was altered by two nucleotides in the mutant, causing alteration in amino acids, pI (from 8.94 to 9.19), and predicted protein structure. Meanwhile, cpsD expression and EPS production were also reduced in the variant strain (p < 0.05) compared with WT, and the exogenous WT-EPS in the variant strain reduced its acid-resistant ability. These results suggested EPS was related to acid responses of BBMN68.
Keywords
Bifidobacterium; adaptive evolution; genome; mutant; exopolysaccharide;
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