地衣芽孢杆菌作为一种嗜碱和降解氰化物微生物的呼吸链改造。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-11-05 DOI:10.1007/s10863-024-10041-y
Daniel Uribe-Ramírez, Lucero Romero-Aguilar, Héctor Vázquez-Meza, Eliseo Cristiani-Urbina, Juan Pablo Pardo
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引用次数: 0

摘要

地衣芽孢杆菌可以利用氰化物作为其生长的氮源。然而,地衣芽孢杆菌在这种化合物(哺乳动物细胞色素 c 氧化酶的典型抑制剂)存在的情况下也能进行有氧呼吸,这表明地衣芽孢杆菌有一条带有多种末端氧化酶的分支呼吸链。在此,我们研究了地衣芽孢杆菌在营养肉汤、含铵的碱性培养基或含氰化物的碱性培养基中培养细胞时呼吸链的变化。然后,我们测量了完整细胞和细胞膜的耗氧量、酶活性,进行了一维和二维-BN-PAGE 分析,并对与 NADH、NADPH 和琥珀酸脱氢酶活性相关的 BN-PAGE 条带进行了质谱分析。我们发现,细胞在营养培养基中比在含有氰化物的碱性培养基中更容易生长。同时,在富营养化培养基和含铵的碱性培养基中培养的细胞呼吸活性逐渐降低,而在含氰的碱性培养基中生长的细胞呼吸活性最低。地衣芽孢杆菌膜含有 NADH、NADPH 和琥珀酸脱氢酶,蛋白质组分析检测到硝酸还原酶和 bc、caa3、aa3 和 bd 复合物。琥珀酸脱氢酶迁移的分子质量为 375 kDa,表明它与硝酸还原酶(分别为 115 kDa + 241 kDa)有关联。地衣芽孢杆菌的 NADH 脱氢酶形成了不同分子质量的聚集体。
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Modifications of the respiratory chain of Bacillus licheniformis as an alkalophilic and cyanide-degrading microorganism.

Bacillus licheniformis can use cyanide as a nitrogen source for its growth. However, it can also carry out aerobic respiration in the presence of this compound, a classic inhibitor of mammalian cytochrome c oxidase, indicating that B. licheniformis has a branched respiratory chain with various terminal oxidases. Here, we studied the modifications in the respiratory chain of B. licheniformis when cells were cultured in Nutrient Broth, an alkaline medium with ammonium, or an alkaline medium with cyanide. Then, we measured oxygen consumption in intact cells and membranes, enzyme activities, carried out 1D and 2D-BN-PAGE, followed by mass spectrometry analysis of BN-PAGE bands associated with NADH, NADPH, and succinate dehydrogenase activities. We found that cell growth was favored in a nutrient medium than in an alkaline medium with cyanide. In parallel, respiratory activity progressively decreased in cells cultured in the rich medium, alkaline medium with ammonium, and the lowest activity was in the cells growing in the alkaline medium with cyanide. B. licheniformis membranes contain NADH, NADPH, and succinate dehydrogenases, and the proteomic analysis detected the nitrate reductase and the bc, caa3, aa3, and bd complexes. The succinate dehydrogenase migrated with a molecular mass of 375 kDa, indicating its association with the nitrate reductase (115 kDa + 241 kDa, respectively). The NADH dehydrogenase of B. licheniformis forms aggregates of different molecular mass.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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