Functional characterization of the phosphotransferase system in Parageobacillus thermoglucosidasius

Abstract

Parageobacillus thermoglucosidasius is a thermophilic bacterium characterized by rapid growth, low nutrient requirements, and amenability to genetic manipulation. These characteristics along with its ability to ferment a broad range of carbohydrates make P. thermoglucosidasius a potential workhorse in whole-cell biocatalysis. The phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) catalyzes the transport and phosphorylation of carbohydrates and sugar derivatives in bacteria, making it important for their physiological characterization. In this study, the role of PTS elements on the catabolism of PTS and non-PTS substrates was investigated for P. thermoglucosidasius DSM 2542. Knockout of the common enzyme I, part of all PTSs, showed that arbutin, cellobiose, fructose, glucose, glycerol, mannitol, mannose, N-acetylglucosamine, N-acetylmuramic acid, sorbitol, salicin, sucrose, and trehalose were PTS-dependent on translocation and coupled to phosphorylation. The role of each putative PTS was investigated and six PTS-deletion variants could not grow on arbutin, mannitol, N-acetylglucosamine, sorbitol, and trehalose as the main carbon source, or showed diminished growth on N-acetylmuramic acid. We concluded that PTS is a pivotal factor in the sugar metabolism of P. thermoglucosidasius and established six PTS variants important for the translocation of specific carbohydrates. This study lays the groundwork for engineering efforts with P. thermoglucosidasius towards efficient utilization of diverse carbon substrates for whole-cell biocatalysis.

Genome sequencing of drake semen micobiome with correlation with their compositions, sources and potential mechanisms affecting semen quality

“…In addition, functional prediction analysis of microbes showed that there were statistical differences in the activities of starch and sucrose metabolism, phosphotransferase system, ABC transporters, microbial metabolism in diverse environments, and quorum sensing pathways between cluster 1 and cluster 2. A previous study reported that the phosphotransferase system could catalyze the transport and phosphorylation of carbohydrates in bacteria ( Bidart et al, 2023 ); ABC transporters were the key carriers of carbohydrate transport and metabolism ( Chandravanshi et al, 2019 ); and quorum sensing was a cell density-dependent gene regulation system that played an important role in carbohydrate metabolism ( Zeng et al, 2021 ). All results of the present study suggest that Chelativorans, Devosia, Halomonas , and Oceanicaulis might affect the semen quality of drakes by regulating carbohydrate metabolism.…”

Sugar transport in thermophiles: Bridging lignocellulose deconstruction and bioconversion

“…In P. thermoglucosidasius , deletions of PTS components confirmed PTS-dependent translocation of common lignocellulosic sugars (cellobiose, fructose, glucose, mannose, sucrose, and trehalose) (Bidart et al., 2023 ). Deletion of Enzyme I in a ∆ ptsI strain eliminated growth on these sugars, while normal growth persisted on maltose and xylose thereby indicating alternative transport mechanisms (Bidart et al., 2023 ). Although the ∆ ptsI strain maintained xylose growth, subsequent deletion of xylose-specific EIIB abolished growth on xylose, revealing the continued dependence on facilitated diffusion catalyzed by the xylose-specific enzyme II despite the absence of phosphorylation (Bidart et al., 2023 ).…”

“…PTS transporters have been studied in two thermophiles: Parageobacillus thermoglucosidasius and Thermoanaerobacter saccharolyticum (Tsakraklides et al., 2012 ; Bidart et al., 2023 ). In P. thermoglucosidasius , deletions of PTS components confirmed PTS-dependent translocation of common lignocellulosic sugars (cellobiose, fructose, glucose, mannose, sucrose, and trehalose) (Bidart et al., 2023 ).…”

“…PTS transporters have been studied in two thermophiles: Parageobacillus thermoglucosidasius and Thermoanaerobacter saccharolyticum (Tsakraklides et al., 2012 ; Bidart et al., 2023 ). In P. thermoglucosidasius , deletions of PTS components confirmed PTS-dependent translocation of common lignocellulosic sugars (cellobiose, fructose, glucose, mannose, sucrose, and trehalose) (Bidart et al., 2023 ). Deletion of Enzyme I in a ∆ ptsI strain eliminated growth on these sugars, while normal growth persisted on maltose and xylose thereby indicating alternative transport mechanisms (Bidart et al., 2023 ).…”

“…The superfamily of ABC sugar transporters are divided into the carbohydrate uptake transporter (CUT) 1 and 2 families, and are collectively responsible for the transport of the broad range of monosaccharides and oligosaccharides available for microbial fermentation following lignocellulose degradation (Schneider, 2001 ; Vanfossen et al., 2009 ). Moreover, the phosphoenolpyruvate-dependent phosphotransferase (PTS) sugar transporter system typically mediates carbon catabolite repression in mesophilic hosts (Warner & Lolkema, 2003 ; Deutscher et al., 2006 , 2014 ; Görke & Stülke, 2008 ); however, there have been comparatively few studies on its influence in thermophiles (Saier & Reizer, 1994 ; Lengeler & Jahreis, 2009 ; Pickl et al., 2012 ; McCoy et al., 2015 ; Bidart et al., 2023 ).…”