Yufang GU, Xiaowei FANG, Chen WANG, Xiongyan LIANG, Chun FANG, Yuying YANG, Xueyang CHEN, Weihuan FANG

Asidik Koşullar Altında Listeria monocytogenes’in Glutamat Dekarboksilazlarının Asit Direnç Sistemlerine Katkılarının Değerlendirilmesi ve Farklı Rollerinin Araştırılması

Listeria monocytogenes, duyarlı insan ve hayvanlarda yüksek ölüm oranı ile seyreden bulaşıcı hastalıklara neden olabilen, önemli bir gıda kaynaklı zoonotik patojendir. L. monocytogenes doğal ortamlarda yaygın olarak bulunduğundan, gıda işleme tesislerinin ve konakçı tarafından tüketilen ürünlerin bakteri ile kontaminasyonu kolaydır. Ancak bir saprofitten hücre içi patojene dönüşmesi sırasında, L. monocytogenes’in karşılaştığı en büyük güçlüklerden biri asit stresidir. Asidik ortamlarla savaşmak için, bakteri, asit tolerans yanıtı (ATR), F0F1-ATPase, glutamat dekarboksilaz (GAD), arginin deiminaz (ADI) ve agmatin deiminaz (AgDI) dahil olmak üzere çeşitli asit direnç sistemleri geliştirmiştir. Bu çalışmada, farklı asit direnç sistemlerinin katkıları kapsamlı bir şekilde değerlendirildi ve üç GAD bileşeninin asidik koşullar altında farklı rolleri araştırıldı. GAD sistemindeki GadD2’nin, L. monocytogenes’in yapay mide sıvısı (AGJ) ve asidik beyin kalp infüzyonunda (BHI) hayatta kalmasına en büyük katkıyı yaptığı ve bunu GAD sisteminden global stres regülatörü SigB, GadD3 ile AgDI sisteminden AguA1 ve ADI sisteminden ArcA’nın izlediği belirlendi. Transkripsiyon analizi, üç GAD’nin mRNA seviyesinin, asit direncine katkıları ile tutarlı olduğunu gösterdi. Benzer sonuçlar, diğer üç temsilci suş olan EGDe, Lm850658 ve M7’de de gözlendi. Ayrıca, üç GAD’nin katkısının, L. monocytogenes’teki protein seviyeleri tarafından belirlendiğini göstermek için saflaştırılmış GAD’ler ve bunların poliantikorlarını elde ettik. GAD sisteminin farklı ekspresyonlarının düzenlenme mekanizmasının anlaşılabilmesi için daha fazla çalışmaya ihtiyaç vardır.

Evaluating the Contribution of Acid Resistance Systems and Probing the Different Roles of the Glutamate Decarboxylases of Listeria monocytogenes Under Acidic Conditions

Listeria monocytogenes is an important zoonotic foodborne pathogen, which can cause a severe invasive illness to susceptible humans and animals with high mortality. As L. monocytogenes is widely distributed in natural environments, the bacterium is easy to contaminate food processing facilities and the productsto be ingested by host. But during the transition from a saprophyte to intracellular pathogen, one of the biggest challenge L. monocytogenes encounters is the acid stress. To combat the acidic environments, the bacterium developed several acid resistance systems, including acid tolerance response (ATR), F0F1-ATPase, glutamate decarboxylase (GAD), arginine deiminase (ADI) and agmatine deiminase (AgDI). In this study, we comprehensively evaluated the contributions of different acid resistance systems and explored the different roles of the three GAD components under acidic conditions. We found that the GadD2 of GAD system made the largest contribution to the survival of L. monocytogenes in artificial gastric juice (AGJ) and acidic brain heart infusion (BHI), which was followed by the global stress regulator SigB, GadD3 of GAD system, AguA1 of AgDI system and ArcA of ADI system. Transcription analysis showed that the mRNA level of the three GADs were consistent with their contribution to acid resistance. Similar results were observed in the other three representative strains EGDe, Lm850658 and M7. We further obtained the purified GADs and their poly-antibodies to demonstrate that the contribution of the three GADs were determined by the protein levels in L. monocytogenes. Further studies are needed to focus on the regulation of different expression of the GAD system.

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