Phosphoenolpyruvate phosphotransferase system components positively regulate Klebsiella biofilm formation

Horng, Yu-Tze; Wang, Chi-Jane; Chung, Wen-Ting; Chao, Huei-Jen; Chen, Yih-Yuan; Soo, Po-Chi

doi:10.1016/j.jmii.2017.01.007

Cited by 24 publications

(24 citation statements)

References 37 publications

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“…We observed reduced 1,3-PD production in this study ( Figure 3A ) and increased capsular polysaccharide production in the previous study ( Horng et al, 2017 ) in K. pneumoniae overexpressing KPN00353-KPN00352 - KPN00351 incubated under microaerobic conditions compared with that of the vector control. This finding is consistent with the study by Guo et al (2010) reporting that non-capsulated K. pneumoniae provided a higher 1,3-PD yield.…”

Section: Resultssupporting

confidence: 71%

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The Negative Effects of KPN00353 on Glycerol Kinase and Microaerobic 1,3-Propanediol Production in Klebsiella pneumoniae

et al. 2017

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1,3-Propanediol (1,3-PD) is a valuable chemical intermediate in the synthesis of polyesters, polyethers, and polyurethanes, which have applications in various products such as cloth, bottles, films, tarpaulins, canoes, foam seals, high-resilience foam seating, and surface coatings. Klebsiella pneumoniae can produce 1,3-PD from glycerol. In this study, KPN00353, an EIIA homologue in the phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS), was found to play a negative regulatory role in 1,3-PD production under microaerobic conditions via binding to glycerol kinase (GlpK). The primary sequence of KPN00353 is similar to those of the fructose-mannitol EIIA (EIIFru and EIIAMtl) family. The interaction between KPN00353 and GlpK resulted in inhibition of the synthesis of glycerol-3-phosphate (G3P) and correlated with reductions in glycerol uptake and the production of 1,3-PD. Based on structure modeling, we conclude that residue H65 of KPN00353 plays an important role in the interaction with GlpK. We mutated this histidine residue to aspartate, glutamate, arginine and glutamine to assess the effects of each KPN00353 variant on the interaction with GlpK, on the synthesis of G3P and on the production of 1,3-PD. Our results illuminate the role of KPN00353 in 1,3-PD production by K. pneumoniae under microaerobic conditions.

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Section: Resultssupporting

confidence: 71%

“…KPN00353 encodes 147-amino-acid (aa) putative EIIA homologue. KPN00352 and KPN00351 encode putative EIIB (95 aa) and EIIC (445 aa) homologues respectively ( Horng et al, 2017 ). Therefore, the purpose of this study was to examine the effects of KPN00353-KPN00352 - KPN00351 on the 1,3-PD production.…”

Section: Introductionmentioning

confidence: 99%

The Negative Effects of KPN00353 on Glycerol Kinase and Microaerobic 1,3-Propanediol Production in Klebsiella pneumoniae

et al. 2017

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“…Our previous data showed that overexpression of KPN00353-KPN00352-KPN00351 increased biofilm formation by K. pneumoniae MGH78578 (Horng et al, 2018). In STU1, we found homologs having high sequence identity to KPN00353 (98%), KPN00352 (100%), and KPN00351 (99%) of MGH78578, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…However, these three assays in the report by Luo et al (2017) did not allow direct observation of type 3 fimbriae synthesis such as Western blotting using antibody specific to type 3 fimbriae. Some other factors in K. pneumoniae may affect the results of MRHA (Stahlhut et al, 2012), mannan-binding assay (Madison et al, 1994), and bacterial adhesion assay (Chung, 2016; Horng et al, 2018). Therefore, the regulation of MRHA, mannan-binding assay and bacterial adhesion assay by KP1-4563 depending on type 3 fimbriae or not is unclear.…”

Section: Discussionmentioning

confidence: 99%

“…We previously found the putative EII complex – KPN00353 (EIIA homolog), KPN00352 (EIIB homolog), and KPN00351 (EIIC homolog) – in the genome of K. pneumoniae MGH78578 (Jeng et al, 2017; Horng et al, 2018). We found the etcA , etcB , and etcC ( etc for enzyme two complex), homologous to KPN00353 , KPN00352 , and KPN00351 , respectively, in K. pneumoniae STU1 in this study.…”

Section: Introductionmentioning

confidence: 99%

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EtcABC, a Putative EII Complex, Regulates Type 3 Fimbriae via CRP-cAMP Signaling in Klebsiella pneumoniae

et al. 2019

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Biofilm formation by Klebsiella pneumoniae on indwelling medical devices increases the risk of infection. Both type 1 and type 3 fimbriae are important factors in biofilm formation by K. pneumoniae . We found that a putative enzyme II (EII) complex of the phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS), etcA (EIIA)- etcB (EIIB)- etcC (EIIC), regulated biofilm and type 3 fimbriae formation by K. pneumoniae STU1. In this study, the regulatory mechanism of etcABC in K. pneumoniae type 3 fimbriae formation was investigated. We found via quantitative RT-PCR that overexpression of etcABC enhanced the transcription level of the mrk operon, which is involved in type 3 fimbriae synthesis, and reduced the transcription level of the fim operon, which is involved in type 1 fimbriae synthesis. To gain further insight into the role of etcABC in type 3 fimbriae synthesis, we analyzed the region upstream of the mrk operon and found the potential cyclic 3′5′-adenosine monophosphate (cAMP) receptor protein (CRP) binding site. After crp was deleted in K. pneumoniae STU1 and two clinical isolates, these three crp mutant strains could not express MrkA, the major subunit of the fimbrial shaft, indicating that CRP positively regulated type 3 fimbriae synthesis. Moreover, a crp mutant overexpressing etcABC could not express MrkA, indicating that the regulation of type 3 fimbriae by etcABC was dependent on CRP. In addition, deletion of cyaA , which encodes the adenylyl cyclase that synthesizes cAMP, and deletion of crr , which encodes the glucose-specific EIIA, led to a reduction in lac operon regulation and therefore bacterial lactose uptake in K. pneumoniae . Exogenous cAMP but not etcABC overexpression compensated for the role of cyaA in bacterial lactose uptake. However, either etcABC overexpression or exogenous cAMP compensated for the role of crr in bacterial lac operon regulation that would eventually restore lactose uptake. We also found via ELISA and the luxCDABE reporter system that overexpression of etcABC increased intracellular cAMP levels and the transcription level of crp , respectively, in K. pneumoniae . In conclusion, overexpression of etcABC positively regulated cAMP production and cAMP-CRP activity to activate the mrk oper...

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Reduction of gut microbial diversity and short chain fatty acids in BALB/c mice exposure to microcystin-LR

Zhang

et al. 2020

Ecotoxicology

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Phosphoenolpyruvate phosphotransferase system components positively regulate Klebsiella biofilm formation

Cited by 24 publications

References 37 publications

The Negative Effects of KPN00353 on Glycerol Kinase and Microaerobic 1,3-Propanediol Production in Klebsiella pneumoniae

The Negative Effects of KPN00353 on Glycerol Kinase and Microaerobic 1,3-Propanediol Production in Klebsiella pneumoniae

EtcABC, a Putative EII Complex, Regulates Type 3 Fimbriae via CRP-cAMP Signaling in Klebsiella pneumoniae

Reduction of gut microbial diversity and short chain fatty acids in BALB/c mice exposure to microcystin-LR

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