TY - JOUR

T1 - The impact of PrsA over-expression on the Bacillus subtilis transcriptome during fed-batch fermentation of alpha-amylase production

AU - Geissler, Adrian S.

AU - Poulsen, Line D.

AU - Doncheva, Nadezhda T.

AU - Anthon, Christian

AU - Seemann, Stefan E.

AU - González-Tortuero, Enrique

AU - Breüner, Anne

AU - Jensen, Lars J.

AU - Hjort, Carsten

AU - Vinther, Jeppe

AU - Gorodkin, Jan

N1 - Publisher Copyright: Copyright © 2022 Geissler, Poulsen, Doncheva, Anthon, Seemann, González-Tortuero, Breüner, Jensen, Hjort, Vinther and Gorodkin.

PY - 2022

Y1 - 2022

N2 - The production of the alpha-amylase (AMY) enzyme in Bacillus subtilis at a high rate leads to the accumulation of unfolded AMY, which causes secretion stress. The over-expression of the PrsA chaperone aids enzyme folding and reduces stress. To identify affected pathways and potential mechanisms involved in the reduced growth, we analyzed the transcriptomic differences during fed-batch fermentation between a PrsA over-expressing strain and control in a time-series RNA-seq experiment. We observe transcription in 542 unannotated regions, of which 234 had significant changes in expression levels between the samples. Moreover, 1,791 protein-coding sequences, 80 non-coding genes, and 20 riboswitches overlapping UTR regions of coding genes had significant changes in expression. We identified putatively regulated biological processes via gene-set over-representation analysis of the differentially expressed genes; overall, the analysis suggests that the PrsA over-expression affects ATP biosynthesis activity, amino acid metabolism, and cell wall stability. The investigation of the protein interaction network points to a potential impact on cell motility signaling. We discuss the impact of these highlighted mechanisms for reducing secretion stress or detrimental aspects of PrsA over-expression during AMY production.

AB - The production of the alpha-amylase (AMY) enzyme in Bacillus subtilis at a high rate leads to the accumulation of unfolded AMY, which causes secretion stress. The over-expression of the PrsA chaperone aids enzyme folding and reduces stress. To identify affected pathways and potential mechanisms involved in the reduced growth, we analyzed the transcriptomic differences during fed-batch fermentation between a PrsA over-expressing strain and control in a time-series RNA-seq experiment. We observe transcription in 542 unannotated regions, of which 234 had significant changes in expression levels between the samples. Moreover, 1,791 protein-coding sequences, 80 non-coding genes, and 20 riboswitches overlapping UTR regions of coding genes had significant changes in expression. We identified putatively regulated biological processes via gene-set over-representation analysis of the differentially expressed genes; overall, the analysis suggests that the PrsA over-expression affects ATP biosynthesis activity, amino acid metabolism, and cell wall stability. The investigation of the protein interaction network points to a potential impact on cell motility signaling. We discuss the impact of these highlighted mechanisms for reducing secretion stress or detrimental aspects of PrsA over-expression during AMY production.

KW - alpha-amylase

KW - Bacillus subtilis

KW - enzyme produced by microorganism

KW - PrsA

KW - RNA sequencing (RNA-seq)

U2 - 10.3389/fmicb.2022.909493

DO - 10.3389/fmicb.2022.909493

M3 - Journal article

C2 - 35992681

AN - SCOPUS:85136486443

VL - 13

JO - Frontiers in Microbiology

JF - Frontiers in Microbiology

SN - 1664-302X

M1 - 909493

ER -