Survival strategy of intestinal lactic acid bacteria in the gut : functional analysis of cell surface structure involved in bile acid adaptation

肠道乳酸菌在肠道中的生存策略：参与胆汁酸适应的细胞表面结构的功能分析

• 批准号: 21380053
• 负责人: YOKOTA Atsushi
• 金额: $ 12.15万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21380053/
• 关键词: 微生物機能; 乳酸菌; 胆汁酸; コール酸; Lactobacillus gasseri; リン脂質; 糖脂質; カルジオリピン; テイコ酸

Free bile acids (FBAs) are considered the most deleterious stress compounds that inhibit the growth of intestinal bacteria by their membrane-damaging effect. Previously, we found adaptation in Lactobacillus gasseri JCM1131T to cholic acid (CA), one of the major FBAs in human intestine. In this adaptation, exponentially growing cells were first exposed to 4 mM CA, a sub-lethal concentration, for 30 min and then to lethal concentration at 15 mM. The adapted cells did not show any appreciable decrease in viability in the presence of 15 mM CA even after 7 h, while non-adapted cells rapidly lost viability by the factors of 10-3~-4. In the adapted cells a significant relief of membrane damage was observed in the presence of CA, suggesting alterations in lipid composition in the cell envelope.Total lipids extracted from adapted and non-adapted cells were separated into three fractions, neutral lipids, glycolipids and phospholipids. Determination of amount of each fraction revealed no signific … More ant changes between adapted and non-adapted cells. Likewise, significant differences were neither found in fatty acids compositions of each fraction. However, significant alterations were detected in glycolipids and phospholipids compositions. In glycolipids, increases in the length of sugar chain were detected during the adaptation. Furthermore, a significant increase in relative amount of cardiolipin (CL) was observed in the phospholipid fraction.To evaluate physiological importance of CL in CA adaptation, vesicle experiment was conducted. It was found that CL confers resistance to the liposome against CA attack, suggesting a pivotal role of CL in adaptation mechanism.Mutants having deletion in two putative cardiolipin synthase genes (cls) singly or in combination were then derived, and their abilities in CA adaptation were investigated. Although CL levels of the both single-knockout mutants were lower than that of the wild type JCM1131T, the levels were found to increase after exposure to 4 mM CA, and a comparable CA adaptation to the wild type strain was observed in both the mutants. The double-knockout mutant possessed negligible amount of CL but still showed CA adaptation, suggesting a dispensable role of CL in CA adaptation. However, in this mutant, significant decrease in glycolipid fraction and significant increase in phospholipid fraction were observed as compared to wild-type strain and these shifts were more prominent after CA adaptation. Therefore, CL was found to work as a key determinant for the lipid composition of the bacterial cell membrane. Less

游离胆汁酸（FBA）被认为是最有害的应激化合物，其通过其膜损伤作用抑制肠道细菌的生长。先前，我们发现加氏乳杆菌JCM 1131 T适应胆酸（CA），这是人类肠道中的主要FBA之一。在这种适应中，指数生长的细胞首先暴露于4 mM CA（亚致死浓度）30 min，然后暴露于15 mM CA的致死浓度。在15 mM CA存在下，适应的细胞即使在7 h后也没有表现出任何明显的存活率下降，而非适应的细胞迅速丧失存活率，丧失的因子为10-3~-4。在适应的细胞中，观察到的CA的存在下，膜损伤的显着救济，这表明在细胞envelope.Total脂质组合物中的改变，从适应和非适应的细胞中提取的被分离成三个馏分，中性脂质，糖脂和磷脂。测定各组分的含量， ...更多信息 蚂蚁在适应和非适应细胞之间变化。同样，各馏分的脂肪酸组成也无显著差异。然而，糖脂和磷脂成分中检测到显着的变化。在糖脂中，在适应过程中检测到糖链长度的增加。此外，心磷脂（CL）的相对量显着增加，观察到的磷脂馏分。结果表明，CL能抵抗脂质体对CA的攻击，表明CL在适应机制中起着关键作用，因此我们获得了两个假定的心磷脂合酶基因（cls）单独或联合缺失的突变体，并研究了它们在CA适应中的能力。尽管两种单敲除突变体的CL水平均低于野生型JCM 1131 T，但发现暴露于4 mM CA后水平增加，并且在两种突变体中观察到对野生型菌株的可比CA适应。双敲除突变体具有可忽略的CL量，但仍显示CA适应，表明CL在CA适应中的调节作用。然而，在这个突变体中，糖脂部分的显着减少和磷脂部分的显着增加，观察到相比，野生型菌株，这些变化后CA适应更加突出。因此，CL被认为是细菌细胞膜脂质组成的关键决定因素。少

项目成果

Lactobacillus gasseri JCM1131Tのリポテイコ酸:胆汁酸適応への関与

加氏乳杆菌 JCM1131T 中的脂磷壁酸：参与胆汁酸适应

• 发表时间: 2012
• 作者: Kouzai Y;Mochizuki S;Saito A;Ando A;Minami E;Nishizawa Y;鈴木健策;白石宗,冨田理,田中尚人,吹谷智,森田直樹,横田伸一,岡田早苗,横田篤
• 通讯作者: 白石宗,冨田理,田中尚人,吹谷智,森田直樹,横田伸一,岡田早苗,横田篤

Bile Acid is a host facor that regulated the composition of rat cecal microbiota

胆汁酸是调节大鼠盲肠微生物群组成的宿主因子

• 发表时间: 2011
• 作者: K.B.M. Saiful Islam;Masahito Hagio;Nobuyuki Fujii;Satoshi Ishizuka;Satoru Fukiya;Atsushi Yokota
• 通讯作者: Atsushi Yokota

乳酸菌・ビフィズス菌における胆汁酸ストレス応答

乳酸菌和双歧杆菌的胆汁酸应激反应

• 发表时间: 2010
• 期刊: Japanese Journal of Lactic Acid Bacteria
• 作者: 鈴木健策;大森幸美;横田篤
• 通讯作者: 横田篤

コール酸添加食摂取がラット盲腸内の細菌叢および胆汁酸代謝に与える影響s

添加胆酸饮食对大鼠盲肠细菌菌群和胆汁酸代谢的影响。

• 发表时间: 2011
• 作者: Yamamoto-Tamura;K.;Ohno M.;Fujii T.;Kataoka;S.;Numata;S.;Nakajima;M.;Hasebe A.;Akutsu K.;slam K.B.M. Saiful,吹谷智,萩尾真人,塚敏,横田篤
• 通讯作者: slam K.B.M. Saiful,吹谷智,萩尾真人,塚敏,横田篤

Lactobacillus gasseri JCM1131^Tにおける推定カルジオリピンシンターゼ遺伝子欠失がコール酸適応現象に与える影響

加氏乳杆菌 JCM1131^T 中假定的心磷脂合酶基因缺失对胆酸适应现象的影响

• 发表时间: 2011
• 作者: 戸部はるひ;澤田真理子;加藤慎二;森田直樹;佐々木泰子;扇谷悟;吹谷智;横田篤
• 通讯作者: 横田篤