Rewiring the Glucose Transportation and Central Metabolic Pathways for Overproduction of N ‐Acetylglucosamine in Bacillus subtilis

2017 ◽  
Vol 12 (10) ◽  
pp. 1700020 ◽  
Author(s):  
Yang Gu ◽  
Jieying Deng ◽  
Yanfeng Liu ◽  
Jianghua Li ◽  
Hyun‐dong Shin ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Pathways

scholarly journals Proteomic responses of spores of Bacillus subtilis to thermosonication involve large-scale alterations in metabolic pathways

2020 ◽  
Vol 64 ◽  
pp. 104992
Author(s):  
Lihua Fan ◽  
Furong Hou ◽  
Aliyu Idris Muhammad ◽  
Balarabe Bilyaminu Ismail ◽  
Ruiling lv ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Pathways ◽  
Large Scale ◽  
Proteomic Responses

scholarly journals Design of a programmable biosensor-CRISPRi genetic circuits for dynamic and autonomous dual-control of metabolic flux in Bacillus subtilis

2019 ◽  
Vol 48 (2) ◽  
pp. 996-1009 ◽  
Author(s):  
Yaokang Wu ◽  
Taichi Chen ◽  
Yanfeng Liu ◽  
Rongzhen Tian ◽  
Xueqin Lv ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Pathways ◽  
Metabolic Flux ◽  
Target Genes ◽  
Fine Tuning ◽  
Dual Control ◽  
Genetic Circuits ◽  
Dynamic Regulation ◽  
Batch Bioreactor ◽  
Acetoin Production

Abstract Dynamic regulation is an effective strategy for fine-tuning metabolic pathways in order to maximize target product synthesis. However, achieving dynamic and autonomous up- and down-regulation of the metabolic modules of interest simultaneously, still remains a great challenge. In this work, we created an autonomous dual-control (ADC) system, by combining CRISPRi-based NOT gates with novel biosensors of a key metabolite in the pathway of interest. By sensing the levels of the intermediate glucosamine-6-phosphate (GlcN6P) and self-adjusting the expression levels of the target genes accordingly with the GlcN6P biosensor and ADC system enabled feedback circuits, the metabolic flux towards the production of the high value nutraceutical N-acetylglucosamine (GlcNAc) could be balanced and optimized in Bacillus subtilis. As a result, the GlcNAc titer in a 15-l fed-batch bioreactor increased from 59.9 g/l to 97.1 g/l with acetoin production and 81.7 g/l to 131.6 g/l without acetoin production, indicating the robustness and stability of the synthetic circuits in a large bioreactor system. Remarkably, this self-regulatory methodology does not require any external level of control such as the use of inducer molecules or switching fermentation/environmental conditions. Moreover, the proposed programmable genetic circuits may be expanded to engineer other microbial cells and metabolic pathways.

scholarly journals Clp-Dependent Proteolysis Down-Regulates Central Metabolic Pathways in Glucose-Starved Bacillus subtilis

2007 ◽  
Vol 190 (1) ◽  
pp. 321-331 ◽  
Author(s):  
Ulf Gerth ◽  
Holger Kock ◽  
Ilja Kusters ◽  
Stephan Michalik ◽  
Robert L. Switzer ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Pathways ◽  
Time Course ◽  
Polyacrylamide Gel Electrophoresis ◽  
Nucleotide Metabolism ◽  
Clp Proteases ◽  
In The Wild ◽  
Branched Chain ◽  
Expression Program ◽  
Intracellular Proteolysis

ABSTRACT Entry into stationary phase in Bacillus subtilis is linked not only to a redirection of the gene expression program but also to posttranslational events such as protein degradation. Using 35S-labeled methionine pulse-chase labeling and two-dimensional polyacrylamide gel electrophoresis we monitored the intracellular proteolysis pattern during glucose starvation. Approximately 200 protein spots diminished in the wild-type cells during an 8-h time course. The degradation rate of at least 80 proteins was significantly reduced in clpP, clpC, and clpX mutant strains. Enzymes of amino acid and nucleotide metabolism were overrepresented among these Clp substrate candidates. Notably, several first-committed-step enzymes for biosynthesis of aromatic and branched-chain amino acids, cell wall precursors, purines, and pyrimidines appeared as putative Clp substrates. Radioimmunoprecipitation demonstrated GlmS, IlvB, PurF, and PyrB to be novel ClpCP targets. Our data imply that Clp proteases down-regulate central metabolic pathways upon entry into a nongrowing state and thus contribute to the adaptation to nutrient starvation. Proteins that are obviously nonfunctional, unprotected, or even “unemployed” seem to be recognized and proteolyzed by Clp proteases when the resources for growth become limited.

scholarly journals Bacillus subtilis Homologs of MviN (MurJ), the Putative Escherichia coli Lipid II Flippase, Are Not Essential for Growth

2009 ◽  
Vol 191 (19) ◽  
pp. 6020-6028 ◽  
Author(s):  
Allison Fay ◽  
Jonathan Dworkin
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Metabolic Pathways ◽  
Growth Defect ◽  
Membrane Translocation ◽  
E Coli ◽  
Peptidoglycan Synthesis ◽  
Lipid Ii

ABSTRACT Although peptidoglycan synthesis is one of the best-studied metabolic pathways in bacteria, the mechanism underlying the membrane translocation of lipid II, the undecaprenyl-disaccharide pentapeptide peptidoglycan precursor, remains mysterious. Recently, it was proposed that the essential Escherichia coli mviN gene encodes the lipid II flippase. Bacillus subtilis contains four proteins that are putatively homologous to MviN, including SpoVB, previously reported to be necessary for spore cortex peptidoglycan synthesis during sporulation. MviN complemented the sporulation defect of a ΔspoVB mutation, and SpoVB and another of the B. subtilis homologs, YtgP, complemented the growth defect of an E. coli strain depleted for MviN. Thus, these B. subtilis proteins are likely to be MviN homologs. However, B. subtilis strains lacking these four proteins have no defects in growth, indicating that they likely do not serve as lipid II flippases in this organism.

Transcriptional profiling of gene expression in response to glucose in Bacillus subtilis: regulation of the central metabolic pathways

2003 ◽  
Vol 5 (2) ◽  
pp. 133-149 ◽  
Author(s):  
Hans-Matti Blencke ◽  
Georg Homuth ◽  
Holger Ludwig ◽  
Ulrike Mäder ◽  
Michael Hecker ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Metabolic Pathways ◽  
Transcriptional Profiling

Improve uridine production by modifying related metabolic pathways in Bacillus subtilis

2020 ◽  
Vol 42 (4) ◽  
pp. 551-555
Author(s):  
Xueting Zhang ◽  
Chen Wang ◽  
Lu Liu ◽  
Rui Ban
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Pathways

scholarly journals Study on Mechanism of γ-PGA and Nattokinase Produced by Bacillus Subtilis Natto Based on RNA-Seq Analysis

2020 ◽  
Author(s):  
Min Li ◽  
Zilong Zhang ◽  
Shenwei Li ◽  
Zhengan Tian ◽  
Xia Ma
Keyword(s):  
Bacillus Subtilis ◽  
Solid State ◽  
Metabolic Pathways ◽  
Fermentation Process ◽  
Molecular Targets ◽  
Commercial Production ◽  
Production Mechanism ◽  
Rna Seq ◽  
Bacillus Subtilis Natto ◽  
Application Prospects

Abstract Poly-γ-glutamic acid (γ-PGA) and nattokinase (NK) are the main substances produced by Bacillus subtilis natto in solid-state fermentation and have wide application prospects. We found that our strains have higher nattokinase activity when soybean is used as a substrate to improve the yield of γ-PGA. Commercial production of γ-PGA and nattokinase requires an understanding of the co-production mechanism. Here, we monitored the metabolites of the fermentation process firstly, analyzed the transcriptome of Bacillus subtilis natto when co-producing γ-PGA and NK, and obtained the maximum γ-PGA yield (11.39 ± 0.38%, w/w) and highest activity of NK during fermentation. By comparing the up-regulated genes and down-regulated genes of key enzymes and product-related metabolic pathways for genetic engineering, the co-production mechanism of γ-PGA and nattokinase can be summarized. This study firstly provides new insights into the co-production mechanism of γ-PGA and nattokinase by Bacillus subtilis natto, and reveals potential molecular targets for promoting the co-production of γ-PGA and nattokinase.

scholarly journals Bacillus subtilis yckG andyckF Encode Two Key Enzymes of the Ribulose Monophosphate Pathway Used by Methylotrophs, and yckH Is Required for Their Expression

1999 ◽  
Vol 181 (23) ◽  
pp. 7154-7160 ◽  
Author(s):  
Hisashi Yasueda ◽  
Yoshio Kawahara ◽  
Shin-ichi Sugimoto
Keyword(s):  
Bacillus Subtilis ◽  
Gene Product ◽  
Gene Structure ◽  
Primary Structure ◽  
Metabolic Pathways ◽  
Sequence Similarity ◽  
Carbon Carbon Bonds ◽  
Detoxification System ◽  
Moderate Sensitivity ◽  
Ribulose Monophosphate Pathway

ABSTRACT The ribulose monophosphate (RuMP) pathway is one of the metabolic pathways for the synthesis of compounds containing carbon-carbon bonds from one-carbon units and is found in many methane- and methanol-utilizing bacteria, which are known as methylotrophs. The characteristic enzymes of this pathway are 3-hexulose-6-phosphate synthase (HPS) and 6-phospho-3-hexuloisomerase (PHI), neither of which was thought to exist outside methylotrophs. However, the presumedyckG gene product (YckG) of Bacillus subtilisshows a primary structure similar to that of methylotroph HPS (F. Kunst et al., Nature 390:249–256, 1997). We have also investigated the sequence similarity between the yckF gene product (YckF) and methylotroph PHI (Y. Sakai, R. Mitsui, Y. Katayama, H. Yanase, and N. Kato, FEMS Microbiol. Lett. 176:125–130, 1999) and found that the yckG and yckF genes of B. subtilis express enzymatic activities of HPS and PHI, respectively. Both of these activities were concomitantly induced inB. subtilis by formaldehyde, with induction showing dependence on the yckH gene, but were not induced by methanol, formate, or methylamine. Disruption of either gene caused moderate sensitivity to formaldehyde, suggesting that these enzymes may act as a detoxification system for formaldehyde in B. subtilis. In conclusion, we found an active yckG (for HPS)-yckF (for PHI) gene structure (now namedhxlA-hxlB) in a nonmethylotroph, B. subtilis, which inherently preserves the RuMP pathway.

scholarly journals Screening and identification of thermophilic cellulolytic bacteria isolated from sawdust compost

2021 ◽  
pp. 348-357
Author(s):  
D. T. H. Phuong ◽  
D. T. Tuyen ◽  
L. V. Thang
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Megaterium ◽  
Metabolic Pathways ◽  
Rrna Gene ◽  
Cellulolytic Bacteria ◽  
16S Rrna Gene Analysis ◽  
Wide Range ◽  
Commercial Enzymes ◽  
Screening And Identification ◽  
Physiological And Biochemical

Composting process mainly depends on the metabolic pathways of the microorganism and involves the activity of different enzymes. Thermophilic cellulase-producing bacteria isolated from sawdust compost were tested for formation of a visible zone around the colonies on the agar plates medium containing carboxymethyl cellulose at 50ºC. Screening of carboxymethyl cellulase producing isolates was further realized on the basis in liquid medium by DNS method. Among 29 isolates investigated, V1 and V11 strains exhibited maximum enzyme activity of 1.9 and 2.3 U/mL, respectively. These isolates were selected for morphological, physiological and biochemical studies and 16S rRNA gene analysis. They were found a Gram-positive, rod-shaped spore forming cells, which were identified as Bacillus megaterium (V1) and Bacillus subtilis (V11) based on cell morphology, nucleotide homology and phylogenetic analysis. The optimal temperature for activity of endoglucanases (CMCase) ranged from 35–45°C (strain V1) and 40– 50ºC (strain V11). Our findings showed that Bacillus megaterium (V1) and Bacillus subtilis (V11) cellulase demonstrate thermophilic characteristics within wide range of temperature and meets the requirements for commercial enzymes. 

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