title: Enhanced Acid Tolerance in Bifidobacterium longum by Adaptive Evolution: Comparison of the Genes between the Acid-Resistant Variant and Wild-Type Strain.
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 10(th), 20(th), 30(th), 40(th), and 50(th) 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 wildtype 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.
authors.affiliation: The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University, Beijing 100083, P.R. China.|Beijing Higher Institution Engineering Research Center of Animal Product, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China.
authors.forename: Yunyun
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University, Beijing 100083, P.R. China.|Beijing Laboratory of Food Quality and Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China.
authors.forename: Fazheng
authors.initials: F
authors.lastname: Ren
authors.affiliation: The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University, Beijing 100083, P.R. China.|Beijing Higher Institution Engineering Research Center of Animal Product, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China.
authors.forename: Songling
authors.initials: S
authors.lastname: Liu
authors.affiliation: The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University, Beijing 100083, P.R. China.|Beijing Higher Institution Engineering Research Center of Animal Product, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University, Beijing 100083, P.R. China.|Key Laboratory of Functional Dairy, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China.
authors.forename: Huiyuan
authors.initials: H
authors.lastname: Guo
authors.affiliation: The Innovation Centre of Food Nutrition and Human Health (Beijing), China Agricultural University, Beijing 100083, P.R. China.|Beijing Laboratory of Food Quality and Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China.
authors.forename: Caiyun
authors.initials: C
authors.lastname: Hou
mesh_terms: D000143:Acids / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D000069978:Bifidobacterium longum / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D005075:Biological Evolution; D015964:Gene Expression Regulation, Bacterial; D006863:Hydrogen-Ion Concentration; D017354:Point Mutation
chemical_list: D000143:Acids; D001426:Bacterial Proteins
keywords: Adaptive Evolution; Bifidobacterium; Exopolysaccharide; Genome; Mutant