Cloning, Expression, and Transcription Analysis of<scp>L</scp>-Arabinose Isomerase Gene from<i>Mycobacterium smegmatis</i>SMDU

Takata, Goro; Poonperm, Wayoon; Rao, Dingqi; Souda, Akane; Nishizaki, Tomoe; Morimoto, Kenji; Izumori, Ken

doi:10.1271/bbb.70177

Cited by 12 publications

(7 citation statements)

References 39 publications

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“…Both top-ranked hits, MSMEG_1704 and MSMEG_1712, are SBPs with an annotated role in carbohydrate transport. MSMEG_1704 belongs to a putative ATP-dependent xylose transporter in close proximity to MSMEG_1712, an SBP potentially involved in arabinose import (Niederweis, 2008;Takata et al, 2007;Titgemeyer et al, 2007). Titgemeyer et al (2007) reported that M. smegmatis grows readily on L-arabinose as the sole carbon source and identified MSMEG_1709-1712 by in silico analysis as an arabinose transporter, naming the corresponding genes of the operon araGFKE.…”

Section: Resultsmentioning

confidence: 99%

Detection and Characterization of a Mycobacterial L-Arabinofuranose ABC Transporter Identified with a Rapid Lipoproteomics Protocol

Müller

Fröhlich

et al. 2019

Cell Chemical Biology

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Section: Resultsmentioning

confidence: 99%

Detection and Characterization of a Mycobacterial L-Arabinofuranose ABC Transporter Identified with a Rapid Lipoproteomics Protocol

Müller

Fröhlich

et al. 2019

Cell Chemical Biology

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“…The utilization of L-arabinose by both E. coli and B. subtilis is highly sensitive to CCR mediated by a global regulator, such as E. coli cyclic AMP receptor protein (23) and B. subtilis CcpA (19). In contrast, L-arabinose-inducible expression of the araA gene in M. smegmatis is not repressed by D-glucose, although the regulatory mechanism is unclear (50). Similar phenomena were observed in the present study, which showed that expression of the L-arabinose regulon of strain ATCC 31831 was not inhibited in the presence of D-glucose ( Table 3), indicating that the LacI-type regulator AraR controls the L-arabinose regulon but the regulatory mechanism is not subject to D-glucose repression.…”

Section: Discussionmentioning

confidence: 99%

“…Among the high-GϩC-content gram-positive bacteria phylogenetically closer to C. glutamicum, on the other hand, a gene cluster for L-arabinose utilization was recently found in two Mycobacterium smegmatis strains, although the genetic arrangements are not identical and the functions of the genes, except araA, have not been determined (50,53).…”

mentioning

confidence: 99%

Identification and Functional Analysis of the Gene Cluster for l -Arabinose Utilization in Corynebacterium glutamicum

Kawaguchi

Sasaki

Vertès

et al. 2009

Appl Environ Microbiol

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Corynebacterium glutamicum ATCC 31831 grew on L-arabinose as the sole carbon source at a specific growth rate that was twice that on D-glucose. The gene cluster responsible for L-arabinose utilization comprised a six-cistron transcriptional unit with a total length of 7.8 kb. Three L-arabinose-catabolizing genes, araA (encoding L-arabinose isomerase), araB (L-ribulokinase), and araD (L-ribulose-5-phosphate 4-epimerase), comprised the araBDA operon, upstream of which three other genes, araR (LacI-type transcriptional regulator), araE (L-arabinose transporter), and galM (putative aldose 1-epimerase), were present in the opposite direction. Inactivation of the araA, araB, or araD gene eliminated growth on L-arabinose, and each of the gene products was functionally homologous to its Escherichia coli counterpart. Moreover, compared to the wild-type strain, an araE disruptant exhibited a >80% decrease in the growth rate at a lower concentration of Larabinose (3.6 g liter Corynebacterium glutamicum is a gram-positive soil microorganism able to secrete large amounts of glutamic acid under suitable conditions (32, 57). Consequently, this organism been used for a long time for industrial production of amino acids and nucleotides, among other primary metabolites (17). Moreover, its potential as a commodity chemical producer is the focus of increasing research efforts (21, 39). Commonly, C. glutamicum is able to grow on various carbon sources, such as hexoses, sugar alcohols, and organic acids (9,29,30). In a future where pentose sugar-rich lignocellulosics should play an important role in global renewable energy, growth of C. glutamicum on pentose sugar-based media is vital to its continued relevance as a producer of commodity chemicals. This has not been possible owing to the lack of essential pentose sugar assimilation pathways in C. glutamicum (4). This contrasts with the characteristics of two major industrial workhorse bacteria, the gram-negative organism Escherichia coli and the grampositive organism Bacillus subtilis, which can grow on L-arabinose as the sole carbon source using the same metabolic pathway. L-Arabinose uptake by E. coli and B. subtilis occurs via two distinct systems involving both a low-affinity H ϩ symporter (encoded by araE) and a high-affinity ATP-dependent transport system (araFGH) (5,18,45). After entering the cell, L-arabinose is sequentially converted to L-ribulose, L-ribulose 5-phosphate, and D-xylulose 5-phosphate by the action of Larabinose isomerase (encoded by araA), L-ribulokinase (araB), and L-ribulose-5-phosphate 4-epimerase (araD), respectively (Fig. 1). D-Xylulose 5-phosphate is eventually metabolized through the reductive pentose phosphate pathway (12). No similar genes for L-arabinose metabolism have been identified in any of the corynebacterial genomes sequenced so far (3,24,38,52,60). Nevertheless, L-arabinose metabolism by C. glutamicum was previously achieved by expression of E. coli araA, araB, and araD genes (26). Although the resulting transformant was able to grow on L-ara...

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“…smegmatis, which is phylogenetically related to C. glutamicum, the L-arabinose catabolic genes are upregulated by L-arabinose (44,45), but the involvement of the AraR homolog in this regulation remains to be investigated. Many other strains of actinobacteria identified to date lack the L-arabinose catabolic genes, but the AraR-dependent regulatory system of L-arabinose utilization reported here might be conserved among other strains that have yet to be unidentified.…”

Section: Figmentioning

confidence: 99%

The LacI-Type Transcriptional Regulator AraR Acts as an l -Arabinose-Responsive Repressor of l -Arabinose Utilization Genes in Corynebacterium glutamicum ATCC 31831

et al. 2014

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bThe Corynebacterium glutamicum ATCC 31831 araBDA operon consists of three L-arabinose catabolic genes, upstream of which the galM, araR, and araE genes are located in opposite orientation. araR encodes a LacI-type transcriptional regulator that negatively regulates the L-arabinose-inducible expression of araBDA and araE (encoding an L-arabinose transporter), through a mechanism that has yet to be identified. Here we show that the AraR protein binds in vitro to three sites: one upstream of araBDA and two upstream of araE. We verify that a 16-bp consensus palindromic sequence is essential for binding of AraR, using a series of mutations introduced upstream of araB in electrophoretic mobility shift assays. Moreover, the DNA-binding activity of AraR is reduced by L-arabinose. We employ quantitative reverse transcription-PCR (qRT-PCR) analyses using various mutant strains deficient in L-arabinose utilization genes to demonstrate that the prominent upregulation of araBDA and araE within 5 min of L-arabinose supplementation is dependent on the uptake but independent of the catabolism of L-arabinose. Similar expression patterns, together with the upregulation by araR disruption without L-arabinose, are evident with the apparent galM-araR operon, although attendant changes in expression levels are much smaller than those realized with the expression of araBDA and araE. The AraR-binding site upstream of araB overlaps the ؊10 region of the divergent galM promoter. These observations indicate that AraR acts as a transcriptional repressor of araBDA, araE, and galM-araR and that L-arabinose acts as an intracellular negative effector of the AraR-dependent regulation.

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Cloning, Expression, and Transcription Analysis ofL-Arabinose Isomerase Gene fromMycobacterium smegmatisSMDU

Cited by 12 publications

References 39 publications

Detection and Characterization of a Mycobacterial L-Arabinofuranose ABC Transporter Identified with a Rapid Lipoproteomics Protocol

Detection and Characterization of a Mycobacterial L-Arabinofuranose ABC Transporter Identified with a Rapid Lipoproteomics Protocol

Identification and Functional Analysis of the Gene Cluster for l -Arabinose Utilization in Corynebacterium glutamicum

The LacI-Type Transcriptional Regulator AraR Acts as an l -Arabinose-Responsive Repressor of l -Arabinose Utilization Genes in Corynebacterium glutamicum ATCC 31831

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