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地衣芽孢杆菌作为嗜碱且能降解氰化物的微生物，其呼吸链的修饰。

Modifications of the respiratory chain of Bacillus licheniformis as an alkalophilic and cyanide-degrading microorganism.

作者信息

Uribe-Ramírez Daniel, Romero-Aguilar Lucero, Vázquez-Meza Héctor, Cristiani-Urbina Eliseo, Pardo Juan Pablo

机构信息

Departamento de Ingeniería Bioquímica, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Av. Wilfrido Massieu s/n, Unidad Profesional Adolfo López Mateos, Gustavo A. Madero, Ciudad de México, 07738, México.

Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Circuito Interior S/N, Ciudad Universitaria, Coyoacán, Ciudad de México, 04510, México.

出版信息

J Bioenerg Biomembr. 2024 Dec;56(6):591-605. doi: 10.1007/s10863-024-10041-y. Epub 2024 Nov 5.

DOI:10.1007/s10863-024-10041-y
PMID:39496989
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11624218/
Abstract

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.

摘要

地衣芽孢杆菌能够利用氰化物作为其生长的氮源。然而，在这种化合物（一种哺乳动物细胞色素c氧化酶的经典抑制剂）存在的情况下，它也能进行有氧呼吸，这表明地衣芽孢杆菌具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时呼吸链的变化。然后，我们测量了完整细胞和细胞膜中的氧气消耗、酶活性，进行了一维和二维蓝 native 聚丙烯酰胺凝胶电泳（1D 和 2D-BN-PAGE），随后对与烟酰胺腺嘌呤二核苷酸（NADH）、烟酰胺腺嘌呤二核苷酸磷酸（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，与在含氰化物的碱性培养基中相比，地衣芽孢杆菌在营养培养基中生长更有利。同时，在丰富培养基、含铵的碱性培养基中培养的细胞呼吸活性逐渐降低，而在含氰化物的碱性培养基中生长的细胞呼吸活性最低。地衣芽孢杆菌的细胞膜含有NADH、NADPH和琥珀酸脱氢酶，蛋白质组学分析检测到了硝酸还原酶以及bc、caa3、aa3和bd复合物。琥珀酸脱氢酶迁移时的分子量为375 kDa，表明它与硝酸还原酶相关（分别为115 kDa和241 kDa）。地衣芽孢杆菌的NADH脱氢酶形成不同分子量的聚集体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/11624218/1ce80709c301/10863_2024_10041_Fig11_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/11624218/869febd503c9/10863_2024_10041_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/11624218/380701f52234/10863_2024_10041_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/11624218/1a073f2c5d8e/10863_2024_10041_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/11624218/2a268b22e456/10863_2024_10041_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea2d/11624218/cccd0fae155c/10863_2024_10041_Fig10_HTML.jpg
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