Life Science Research Center, College of Bioresource Sciences, Nihon University, Kanagawa, Japan.
Mizkan Holdings Co., Ltd, Aichi, Japan.
Sci Rep. 2021 Nov 3;11(1):21567. doi: 10.1038/s41598-021-00998-w.
Acetic acid bacteria (AAB) are industrial microorganisms used for vinegar fermentation. Herein, we investigated the distribution and genome structures of mitomycin C-inducible temperate phages in AAB. Transmission electron microscopy analysis revealed phage-like particles in 15 out of a total 177 acetic acid bacterial strains, all of which showed morphology similar to myoviridae-type phage. The complete genome sequences of the six phages derived from three strains each of Acetobacter and Komagataeibacter strains were determined, harboring a genome size ranging from 34,100 to 53,798 bp. A phage AP1 from A. pasteurianus NBRC 109446 was predicted as an active phage based on the genomic information, and actually had the ability to infect its phiAP1-cured strain. The attachment sites for phiAP1 were located in the 3'-end region of the tRNA gene. We also developed a chromosome-integrative vector, p2096int, based on the integrase function of phiAP1, and it was successfully integrated into the attachment site of the phiAP1-cured strain, which may be used as a valuable tool for the genetic engineering. Overall, this study showed the distribution of mitomycin C-inducible temperate phages in AAB, and identified the active temperate phage o f A. pasteurianus.
醋酸菌(AAB)是用于醋发酵的工业微生物。在此,我们研究了 AAB 中米托霉素诱导的温和噬菌体的分布和基因组结构。透射电子显微镜分析显示,在总共 177 株醋酸细菌菌株中,有 15 株存在噬菌体样颗粒,它们的形态均与肌病毒科噬菌体相似。从三株醋杆菌和三株 komagataeibacter 菌株中各获得了 6 种噬菌体的完整基因组序列,基因组大小从 34100bp 到 53798bp 不等。根据基因组信息,预测来自 A. pasteurianus NBRC 109446 的噬菌体 AP1 为活性噬菌体,实际上它具有感染其 phiAP1 修复株的能力。phiAP1 的附着位点位于 tRNA 基因的 3'-末端区域。我们还基于 phiAP1 的整合酶功能开发了一种染色体整合载体 p2096int,并成功整合到 phiAP1 修复株的附着位点,这可能成为遗传工程的有用工具。总的来说,这项研究显示了 AAB 中米托霉素诱导的温和噬菌体的分布,并鉴定了 A. pasteurianus 的活性温和噬菌体。