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

Jiang, Yunyun; Ren, Fazheng; Liu, Songling; Zhao, Liang; Guo, Huiyuan; Hou, Caiyun

doi: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 원문보기

Journal of microbiology and biotechnology, v.26 no.3, 2016년, pp.452 - 460

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)

Abstract ▼ AI-Helper

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.

주제어

AI 본문요약
AI-Helper

문제 정의

Strain BBMN68 plays vital roles in maintaining digestive function, improving constipation, and increasing the immunologic function of the host, per studies carried out in mice [9,13,31]. This study aimed to improve the acid resistance of BBMN68 through adaptive evolution. The stable resistant mutant of BBMN68 was screened under acid stress.

데이터처리

0 (Chicago, USA). Comparisons were done using one-way analysis of variance, followed by the least significant differences comparison test (set at 5%).

이론/모형

5 kDa Visking dialysis membrane at 4℃, with renewal of water per 12 h. The exopolysaccharide (EPS) concentration was determined using the phenol-sulfuric acid colorimetric method.
longum subsp. longum BBMN68 was obtained using the adaptive evolution method. The EPS change may be the critical factor for the acid tolerance of BBMN68m, which is caused by the cpsD gene.

성능/효과

The mutant E72 from Bacillus subtlis168 exhibited higher growth rate and xylose consumption rate with three mutations (araR, sinR, and comP) [32]. In this study, comparative whole-genome analysis showed that BBMN68m contained 19 mutant genes, of which six genes represented changes in the amino acid sequences. One of the mutant genes participated in EPS synthesis and changed the content of the production, which may be the reason of affecting the acid tolerance in the strains.
A wild-type Lactobacillus crispatus showed a cell aggregation phenotype and had improved gastrointestinal persistence compared with its spontaneous non-aggregating mutant [3]. In this study, the BBMN68m strain was more aggregative and was lower in EPS producing content compared with that of the wild-type strain (Fig. 2).
7). Perhaps the high level of EPS originating from the WT strain itself caused its low aggregation, and the effect of exogenous mutant-EPS was limited. Moreover, there were differences in the main molecular weight and structure but not in monomer composition functional groups (data unpublished).
The EPS change may be the critical factor for the acid tolerance of BBMN68m, which is caused by the cpsD gene. Taken together, the results presented here indicate that we not only obtained an acid-resistant strain for the food industry, but also contributed to revealing the possible mechanisms of acid tolerance of Bifidobacterium.
However, there were no reports that demonstrated the relationship between the structure, monomer composition, molecular weight, and acid tolerance. The results suggest the EPS may be a key influencing factor for the response to acid stress in BBMN68 and BBMN68m strains.

참고문헌 (35)

Alp G, Aslim B. 2010. Relationship between the resistance to bile salts and low pH with exopolysaccharide (EPS) production of Bifidobacterium spp. isolated from infants feces and breast milk. Anaerobe 16: 101-105.

상세보기
Brennan TCR, Williams TC, Schulz BL, Palfreyman RW, Kromer JO, Nielsen LK. 2015. Evolutionary engineering improves tolerance for replacement jet fuels in Saccharomyces cerevisiae. Appl. Environ. Microbiol. 81: 3316-3325.

상세보기
Cesena C, Morelli L, Alander M, Siljander T, Tuomola E, Salminen S, et al. 2001. Lactobacillus crispatus and its nonaggregating mutant in human colonization trials. J. Dairy Sci. 84: 1001-1010.

상세보기
Colvin KM, Gordon VD, Murakami K, Borlee BR, Wozniak DJ, Wong GCL, et al. 2011. The Pel polysaccharide can serve a structural and protective role in the biofilm matrix of Pseudomonas aeruginosa. PloS Pathog. 7: e1001264.

상세보기
Colvin KM, Irie Y, Tart CS, Urbano R, Whitney JC, Ryder C, et al. 2012. The Pel and Psl polysaccharides provide Pseudomonas aeruginosa structural redundancy within the biofilm matrix. Environ. Microbiol. 14: 1913-1928.

상세보기
Fanning S, Hall LJ, Cronin M, Zomer A, MacSharry J, Goulding D, et al. 2012. Bifidobacterial surface-exopolysaccharide facilitates commensal-host interaction through immune modulation and pathogen protection. Proc Natl. Acad. Sci. USA 109: 2108-2113.

상세보기
Fong SS, Joyce AR, Palsson BO. 2005. Parallel adaptive evolution cultures of Escherichia coli lead to convergent growth phenotypes with different gene expression states. Genome Res. 15: 1365-1372.

상세보기
Ginalski K, von Grotthuss M, Grishin NV, Rychlewski L. 2004. Detecting distant homology with Meta-BASIC. Nucleic Acids Res. 32: W576-W581.

상세보기
Hao YL, Huang DW, Guo HY, Xiao M, An HR, Zhao LA, et al. 2011. Complete genome sequence of Bifidobacterium longum subsp. longum BBMN68, a new strain from a healthy Chinese centenarian. J. Bacteriol. 193: 787-788.

상세보기
Hidalgo-Cantabrana, Sanchez C, Milani B, Ventura C, Margolles M, Ruas-Madiedo AP. 2014. Genomic overview and biological functions of exopolysaccharide biosynthesis in Bifidobacterium spp. Appl. Environ. Microbiol. 80: 9-18.

상세보기
Hidalgo-Cantabrana C, Sanchez B, Moine D, Berger B, de los Reyes-Gavilan CG, Gueimonde M, et al. 2013. Insights into the ropy phenotype of the exopolysaccharide-producing strain Bifidobacterium animalis subsp. lactis A1dOxR. Appl. Environ. Microbiol. 79: 3870-3874.

상세보기
Horn N, Wegmann U, Dertli E, Mulholland F, Collins SR, Waldron KW, et al. 2013. Spontaneous mutation reveals influence of exopolysaccharide on Lactobacillus johnsonii surface characteristics. PloS One 8: e59957.

상세보기
Jin JH, Zhang B, Guo HY, Cui J, Jiang L, Song S, et al. 2012. Mechanism analysis of acid tolerance response of Bifidobacterium longum subsp. longum BBMN68 by gene expression profile using RNA-sequencing. PloS One 7: 581-589.
Kolodkin-Gal I, Cao SG, Chai L, Bottcher T, Kolter R, Clardy J, et al. 2012. A self-produced trigger for biofilm disassembly that targets exopolysaccharide. Cell 149: 684-692.

상세보기
Krasteva PV, Giglio KM, Sondermann H. 2012. Sensing the messenger: the diverse ways that bacteria signal through cdi-GMP. Protein Sci. 21: 929-948.

상세보기
Lairson LL, Henrissat B, Davies GJ, Withers SG. 2008. Glycosyltransferases: structures, functions, and mechanisms. Annu. Rev. Biochem. 77: 521-555.

상세보기
Lee JH, Karamychev VN, Kozyavkin SA, Mills D, Pavlov AR, Pavlova NV, et al. 2008. Comparative genomic analysis of the gut bacterium Bifidobacterium longum reveals loci susceptible to deletion during pure culture growth. BMC Genomics 9: 247-263.

상세보기
Ling H, Teo WS, Chen BB, Leong SSJ, Chang MW. 2014. Microbial tolerance engineering toward biochemical production: from lignocellulose to products. Curr. Opin. Biotechnol. 29: 99-106.

상세보기
Margolles A, Sanchez B. 2012. Selection of a Bifidobacterium animalis subsp. lactis strain with a decreased ability to produce acetic acid. Appl. Environ. Microbiol. 78: 3338-3342.

상세보기
Mills S, Stanton S, Fitzgerald C, Ross GF. 2011. Enhancing the stress responses of probiotics for a lifestyle from gut to product and back again. Microb. Cell Fact. 10: S19.

상세보기
Morona J, Paton JC, Miller DC, Morona R. 2000. Tyrosine phosphorylation of CpsD negatively regulates capsular polysaccharide biosynthesis in Streptococcus pneumoniae. Mol. Microbiol. 35: 1431-1442.

상세보기
Pitino I, Randazzo CL, Cross KL, Parker ML, Bisignano C, Wickham MSJ, et al. 2012. Survival of Lactobacillus rhamnosus strains inoculated in cheese matrix during simulated human digestion. Food Microbiol. 31: 57-63.

상세보기
Royce LA, Yoon JM, Chen YX, Rickenbach E, Shanks JV, Jarboe LR. 2015. Evolution for exogenous octanoic acid tolerance improves carboxylic acid production and membrane integrity. Metab. Eng. 29: 180-188.

상세보기
Sanchez B, Champomier-Verges MC, Collado MD, Anglade P, Baraige F, Sanz Y, et al. 2007. Low-pH adaptation and the acid tolerance response of Bifidobactetium longum biotype longum. Appl. Environ. Microbiol. 73: 6450-6459.

상세보기
Serra DO, Hengge R. 2014. Stress responses go three dimensional - the spatial order of physiological differentiation in bacterial macrocolony biofilms. Environ. Microbiol. 16: 1455-1471.

상세보기
Tenaillo O, Rodriguez-Verdugo A, Gaut RL, McDonald P, Bennett AF, Long AD, et al. 2012. The molecular diversity of adaptive convergence. Science 335: 457-461.

상세보기
Wai SN, Mizunoe Y, Takade A, Kawabata S, Yoshida S. 1998. Vibrio cholerae O1 strain TSI-4 produces the exopolysaccharide materials that determine colony morphology, stress resistance, and biofilm formation. Appl. Environ. Microbiol. 64: 3648-3655.

상세보기
Xiao J, Klein MI, Falsetta ML, Lu BW, Delahunty CM, Yates JR, et al. 2012. The exopolysaccharide matrix modulates the interaction between 3D architecture and virulence of a mixed-species oral biofilm. PloS Pathog. 8: e1002623.

상세보기
Xiao M, Xu P, Zhao JY, Wang Z, Zuo FL, Zhang JW, et al. 2011. Oxidative stress-related responses of Bifidobacterium longum subsp. longum BBMN68 at the proteomic level after exposure to oxygen. Microbiology 157: 1573-1588.

상세보기
Yang HX, Deng JJ, Yuan Y, Fan DD, Zhang Y, Zhang RJ, et al. 2015. Two novel exopolysaccharides from Bacillus amyloliquefaciens C-1: antioxidation and effect on oxidative stress. Curr. Microbiol. 70: 298-306.

상세보기
Yang HY, Liu SL, Ibrahim SA, Zhao L, Jiang JL, Sun WF, et al. 2009. Oral administration of live Bifidobacterium substrains isolated from healthy centenarians enhanced immune function in BALB/c mice. Nutr. Res. 29: 281-289.

상세보기
Zhang B, Li N, Wang ZW, Tang YJ, Chen T, Zhao XM. 2015. Inverse metabolic engineering of Bacillus subtilis for xylose utilization based on adaptive evolution and whole-genome sequencing. Appl. Microbiol. Biotechnol. 99: 885-896.

상세보기
Zhang J , Wu CD, Du GC, Chen J. 2012. Enhanced a cid tolerance in Lactobacillus casei by adaptive evolution and compared stress response during acid stress. Biotechnol. Bioprocess Eng. 17: 283-289.

원문보기 상세보기
Zhang L, Liu CH, Li D, Zhao YJ, Zhang X, Zeng XP, et al. 2013. Antioxidant activity of an exopolysaccharide isolated from Lactobacillus plantarum C88. Int. J. Biol. Macromol. 54: 270-275.

상세보기
Zhao L, Qiao X, Zhu J, Zhang X, Jiang J, Hao Y, et al. 2011. Correlations of fecal bacterial communities with age and living region for the elderly living in Bama, Guangxi, China. J. Microbiol. 49: 186-192.

원문보기 상세보기

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증