{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T15:51:42Z","timestamp":1786117902669,"version":"build-2736575974"},"reference-count":39,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22478324"],"award-info":[{"award-number":["22478324"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Scientific research foundation of the state key laboratory of vaccines for infectious diseases, Xiang an biomedicine laboratory","award":["2024XAKJ0100007"],"award-info":[{"award-number":["2024XAKJ0100007"]}]},{"DOI":"10.13039\/501100003392","name":"Natural Science Foundation of Fujian Province","doi-asserted-by":"publisher","award":["2022J05223"],"award-info":[{"award-number":["2022J05223"]}],"id":[{"id":"10.13039\/501100003392","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,8,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>Phospholipase D (PLD) has multiple functions in cells and various industrial applications, including the production of high-purity and rare phospholipids, making it a highly valuable enzyme. However, the strong cytotoxicity of PLD and limited understanding of its mechanism in prokaryotic systems have resulted in low levels of heterologous PLD expression. In this study, we used a PLD-expressing strain model to characterize the morphological and physiological changes in PLD-exposed cells. By analyzing the intracellular ion content and cell membrane potential, we found that PLD-induced phosphatidic acid accumulation abnormally stimulated the continuous efflux of potassium ions through voltage-gated potassium channels. Disrupted ion homeostasis affects cell growth and metabolism, leading to a burst of reactive oxygen species levels and causing cellular damage. Transcriptomic analysis suggests that promoting the conversion of fatty acids to phospholipids, reducing phosphatidic acid accumulation, and minimizing phospholipid composition disruptions are key strategies for alleviating PLD cytotoxicity. This study highlights how PLD directly disrupts phospholipid homeostasis through its catalytic activity or indirectly affects cellular metabolism via phosphatidic acid as a signaling molecule. These findings are crucial for understanding the cellular mechanisms of PLD and enhancing heterologous expression levels.<\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        This study provides valuable insights into the cellular mechanisms of enzyme cytotoxicity, using\n                        <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                        expressing phospholipase D (PLD) as a model. By systematically analyzing physiological changes and transcriptomic profiles, it probes key factors such as ion homeostasis and membrane potential, offering a robust framework for understanding the barriers to heterologous protein expression. The discovery that PLD-induced accumulation of phosphatidic acid triggers continuous potassium efflux via voltage-gated channels underscores the critical link between disrupted ion balance and cell survival. These findings advance our understanding of PLD-mediated toxicity and pave the way for future metabolic engineering strategies designed to enhance the industrial production of this and other high-value enzymes.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03763-25","type":"journal-article","created":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T13:01:54Z","timestamp":1782133314000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Cytotoxicity mechanism of heterologous expression of phospholipase D in\n                    <i>Escherichia coli<\/i>"],"prefix":"10.1128","volume":"14","author":[{"given":"Shaofeng","family":"Chen","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00mcjh785","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemical and Biochemical Engineering, College of 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