Glucose-Mediated Transcriptional Repression of PCB/Biphenyl Catabolic Genes in Rhodococcus jostii RHA1

Araki, Naoto; Niikura, Yuji; Miyauchi, Keisuke; Kasai, Daisuke; Masai, Eiji; Fukuda, Masao

doi:10.1159/000323509

Cited by 12 publications

(8 citation statements)

References 65 publications

(35 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The bacterial strains and plasmids used in this study are listed in Table 1. Rhodococcus jostii RHA1 and its mutant derivatives were routinely grown at 30 °C in Luria-Bertani (LB) medium, 0.2× LB medium, or W minimal salt medium [30] containing 10 or 20 mM n -alkanes (C9 to C30) or 20 mM pyruvate. In solid cultures on W minimal medium agar, n -alkanes (C5 to C13) were supplemented as vapor.…”

Section: Methodsmentioning

confidence: 99%

Characterization and Transcriptional Regulation of n-Alkane Hydroxylase Gene Cluster of Rhodococcus jostii RHA1

et al. 2019

View full text Add to dashboard Cite

Gram-positive actinomycete Rhodococcus jostii RHA1 is able to grow on C10 to C19 n-alkanes as a sole source of carbon and energy. To clarify, the n-alkane utilization pathway—a cluster of 5 genes (alkBrubA1A2BalkU) which appeared to be involved in n-alkane degradation—was identified and the transcriptional regulation of these genes was characterized. Reverse transcription-PCR analyses revealed that these genes constituted an operon and were transcribed in the presence of n-alkane. Inactivation of alkB led to the absence of the ability to utilize n-undecane. The alkB mutation resulted in reduction of growth rates on C10 and C12 n-alkanes; however, growths on C13 to C19 n-alkanes were not affected by this mutation. These results suggested that alkB was essential for the utilization of C10 to C12 n-alkanes. Inactivation of alkU showed the constitutive expression of alkB. Purified AlkU is able to bind to the putative promoter region of alkB, suggesting that AlkU played a role in repression of the transcription of alk operon. The results of this study indicated that alkB was involved in the medium-chain n-alkanes degradation of strain RHA1 and the transcription of alk operon was negatively regulated by alkU-encoded regulator. This report is important to understand the n-alkane degradation pathway of R. jostii, including the transcriptional regulation of alk gene cluster.

show abstract

Section: Methodsmentioning

confidence: 99%

Characterization and Transcriptional Regulation of n-Alkane Hydroxylase Gene Cluster of Rhodococcus jostii RHA1

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Having established the conversion of protocatechuic acid to 2,4-PDCA, the production of 2,4-PDCA was first tested using an aromatic precursor 4-hydroxybenzoic acid as carbon source. Growth of R. jostii pcaHG:ligAB(P tpc5 ) in LB media containing 0.1% 4-hydroxybenzoic acid resulted in efficient conversion to 2,4-PDCA, but growth in M9 minimal media in the presence of 0.1% glucose gave very weak production of 2,4-PDCA, likely due to catabolite repression by glucose [22]. However, growth of this strain using 0.1% yeast extract as media additive in place of glucose was found to generate 2,4-PDCA and PCA from M9/0.1% 4-hydroxybenzoic acid (see Additional file 1: Figure S16).…”

Section: Production Of 24-pdca From Polymeric Ligninmentioning

confidence: 99%

Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin

Spence

Calvo-Bado

Mines³

et al. 2021

Microb Cell Fact

View full text Add to dashboard Cite

Genetic modification of Rhodococcus jostii RHA1 was carried out in order to optimise the production of pyridine-2,4-dicarboxylic acid and pyridine-2,5-dicarboxylic acid bioproducts from lignin or lignocellulose breakdown, via insertion of either the Sphingobium SYK-6 ligAB genes or Paenibacillus praA gene respectively. Insertion of inducible plasmid pTipQC2 expression vector containing either ligAB or praA genes into a ΔpcaHG R. jostii RHA1 gene deletion strain gave 2–threefold higher titres of PDCA production from lignocellulose (200–287 mg/L), compared to plasmid expression in wild-type R. jostii RHA1. The ligAB genes were inserted in place of the chromosomal pcaHG genes encoding protocatechuate 3,4-dioxygenase, under the control of inducible Picl or PnitA promoters, or a constitutive Ptpc5 promoter, producing 2,4-PDCA products using either wheat straw lignocellulose or commercial soda lignin as carbon source. Insertion of Amycolatopsis sp. 75iv2 dyp2 gene on a pTipQC2 expression plasmid led to enhanced titres of 2,4-PDCA products, due to enhanced rate of lignin degradation. Growth in minimal media containing wheat straw lignocellulose led to the production of 2,4-PDCA in 330 mg/L titre in 40 h, with > tenfold enhanced productivity, compared with plasmid-based expression of ligAB genes in wild-type R. jostii RHA1. Production of 2,4-PDCA was also observed using several different polymeric lignins as carbon sources, and a titre of 240 mg/L was observed using a commercially available soda lignin as feedstock.

show abstract

“…Rhodococcus jostii strain RHA1 was grown at 30°C in one-fifth-diluted LB (1/5 LB) broth or in W minimal salt medium (2). W minimal salt medium contains neither nitrate nor nitrite.…”

Section: Methodsmentioning

confidence: 99%

“…DNA manipulations were performed essentially as described by Ausubel et al and Sambrook et al (4,21). Gene deletion mutants were constructed by allelic exchange using sacB counterselection and confirmed by Southern hybridization and nucleotide sequencing (2). Southern hybridization was performed using the DIG system (Boehringer Mannheim Biochemicals, Indianapolis, IN) according to the manufacturer's protocol.…”

Section: Methodsmentioning

confidence: 99%

Specific Gene Responses of Rhodococcus jostii RHA1 during Growth in Soil

Iino

Wang

Miyauchi

et al. 2012

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

ABSTRACTTranscriptome analysis ofRhodococcus jostiiRHA1 during growth in sterilized soil was performed. A total of 165 soil-specific genes were identified by subtracting genes upregulated in late growth phases and on solid medium from 264 genes commonly upregulated during growth on biphenyl or pyruvate in sterilized soil. Classification of the 165 genes into functional categories indicated that this soil-specific group is rich in genes for the metabolism of fatty acids, amino acids, carbohydrates, and nitrogen and relatively poor in those for cellular processes and signaling. The ro06365–ro06369 gene cluster, in which ro06365 to ro06368 were highly upregulated in transcriptome analysis, was characterized further. ro06365 and ro06366 show similarity to a nitrite/nitrate transporter and a nitrite reductase, respectively, suggesting their involvement in nitrogen metabolism. A strain with an ro06366 deletion, D6366, showed growth retardation when we used nitrate as the sole nitrogen source and no growth when we used nitrite. A strain with a deletion of ro06365 to ro06368, DNop, utilized neither nitrite nor nitrate and recovered growth using nitrite and nitrate by introduction of the deleted genes. Both of the mutants showed growth retardation in sterilized soil, and the growth retardation of DNop was more significant than that of D6366. When these mutants were cultivated in medium containing the same proportions of ammonium, nitrate, and nitrite ions as those in the sterilized soil, they showed growth retardation similar to that in the soil. These results suggest that the ro06365–ro06369 gene cluster has a significant role in nitrogen utilization in sterilized soil.

show abstract

Glucose-Mediated Transcriptional Repression of PCB/Biphenyl Catabolic Genes in Rhodococcus jostii RHA1

Cited by 12 publications

References 65 publications

Characterization and Transcriptional Regulation of n-Alkane Hydroxylase Gene Cluster of Rhodococcus jostii RHA1

Characterization and Transcriptional Regulation of n-Alkane Hydroxylase Gene Cluster of Rhodococcus jostii RHA1

Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin

Specific Gene Responses of Rhodococcus jostii RHA1 during Growth in Soil

Contact Info

Product

Resources

About