A<i>Corynebacterium glutamicum rnhA recG</i>Double Mutant Showing Lysozyme- sensitivity, Temperature-sensitive Growth, and UV-Sensitivity

Hirasawa, Takashi; Kumagai, Yutaro; Nagai, Kazuo; Wachi, Masaaki

doi:10.1271/bbb.67.2416

Cited by 13 publications

(11 citation statements)

References 41 publications

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“…The RNase H and APase activities of the full length SCO2299 protein depend on its N-terminal RNase H domain and C-terminal APase domain, respectively, and do not interfere or overlap with each other. Although C. glutamicum RNase HI, the SCO2299-like protein, was shown to be active as an RNase H, its phosphatase activity had not previously been examined [24]. Therefore, the SCO2299 protein is the first reported example of this bifunctional RNase HI.…”

Section: Discussionmentioning

confidence: 99%

“…Previous work has shown that Corynebacterium glutamicum RNase HI (Type 1 RNase H), whose additional C-terminal region showed a high amino acid sequence similarity to the CobC protein, exhibited RNase H activity in vivo in a complementation assay with an E. coli RNase H-deficient strain [24]. Although the RNase HI with the extra CobC-like region is a novel style in the Type 1 RNase H family, the C. glutamicum enzyme itself has not been characterized and a function of its C-terminal region remains unknown [24]. The C-terminal region of the enzyme certainly shows a similarity to the CobC protein, which has been reported to be involved in synthesis of cobalamin, one of the precursors of vitamin B 12 synthesis [25].…”

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confidence: 99%

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The SCO2299 gene from Streptomyces coelicolor A3(2) encodes a bifunctional enzyme consisting of an RNase H domain and an acid phosphatase domain

et al. 2005

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The SCO2299gene from Streptomyces coelicolor encodes a single peptide consisting of 497 amino acid residues. Its N‐terminal region shows high amino acid sequence similarity to RNase HI, whereas its C‐terminal region bears similarity to the CobC protein, which is involved in the synthesis of cobalamin. The SCO2299 gene suppressed a temperature‐sensitive growth defect of an Escherichia coli RNase H‐deficient strain, and the recombinant SCO2299 protein cleaved an RNA strand of RNA·DNA hybrid in vitro. The N‐terminal domain of the SCO2299 protein, when overproduced independently, exhibited RNase H activity at a similar level to the full length protein. On the other hand, the C‐terminal domain showed no CobC‐like activity but an acid phosphatase activity. The full length protein also exhibited acid phosphatase activity at almost the same level as the C‐terminal domain alone. These results indicate that RNase H and acid phosphatase activities of the full length SCO2299 protein depend on its N‐terminal and C‐terminal domains, respectively. The physiological functions of the SCO2299 gene and the relation between RNase H and acid phosphatase remain to be determined. However, the bifunctional enzyme examined here is a novel style in the Type 1 RNase H family. Additionally, S. coelicolor is the first example of an organism whose genome contains three active RNase H genes.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

The SCO2299 gene from Streptomyces coelicolor A3(2) encodes a bifunctional enzyme consisting of an RNase H domain and an acid phosphatase domain

et al. 2005

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“…Database searches using the Sto ‐RNase HI sequence indicate that not only the archaeal genomes, but also the bacterial and eukaryotic genomes contain the genes encoding a Sto ‐RNase HI homologue. Of them, the Sto ‐RNase HI homologue from B. subtilis (YpdQ) is inactive [3], whereas those from Streptomyces coelicolor (SCO7284 [18] and SCO2299 [19]) and Corynebacterium glutamicum (Cg12236) [20] are active. Phylogenetic analyses of the type 1 RNase H sequences show that the Sto ‐RNase HI homologues form an independent domain, which is different from those of bacterial, eukaryotic and retroviral type 1 RNases H [18].…”

Section: Identification Of Archaeal Type 1 Rnases Hmentioning

confidence: 99%

Ribonuclease H: molecular diversities, substrate binding domains, and catalytic mechanism of the prokaryotic enzymes

2009

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The prokaryotic genomes, for which complete nucleotide sequences are available, always contain at least one RNase H gene, indicating that RNase H is ubiquitous in all prokaryotic cells. Coupled with its unique substrate specificity, the enzyme has been expected to play crucial roles in the biochemical processes associated with DNA replication, gene expression and DNA repair. The physiological role of prokaryotic RNases H, especially of type 1 RNases H, has been extensively studied using Escherichia coli strains that are defective in RNase HI activity or overproduce RNase HI. However, it is not fully understood yet. By contrast, significant progress has been made in this decade in identifying novel RNases H with respect to their biochemical properties and structures, and elucidating catalytic mechanism and substrate recognition mechanism of RNase H. We review the results of these studies.

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“…The E. coli udhA gene was amplified by polymerase chain reaction (PCR) from E. coli W3110 genomic DNA using KOD-plus-DNA polymerase and the following primers: 5 -TGGCCGGATCCTTTACGTACAGCGG-3 and 5 -AAGC CGGATCCACGCCGCACTATTGG-3 . The amplified fragment was digested with restriction endonuclease BamHI and then cloned into the BamHI site of vector pHT1 as described previously (Hirasawa et al, 2003). The resulting plasmid was named pHT1-udhA, and was transformed into C. glutamicum ATCC 13032.…”

Section: Introductionmentioning

confidence: 99%

Enhanced acetic acid and succinic acid production under microaerobic conditions by Corynebacterium glutamicum harboring Escherichia coli transhydrogenase gene pntAB

Yamauchi¹,

Hirasawa²,

Nishii³

et al. 2014

J. Gen. Appl. Microbiol.

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Some microorganisms, such as Escherichia coli, harbor transhydrogenases that catalyze the interconversion between NADPH and NADH. However, such transhydrogenase genes have not been found in the genome of a glutamic acid-producing bacterium Corynebacterium glutamicum. In this study, the E. coli transhydrogenase genes udhA and pntAB were introduced into the C. glutamicum wild-type strain ATCC 13032, and the metabolic characteristics of the recombinant strains under aerobic and microaerobic conditions were examined. No major metabolic changes were observed following the introduction of the E. coli transhydrogenase genes under aerobic conditions. Under microaerobic conditions, significant metabolic change was not observed following the introduction of the udhA gene. However, the specific production rates of lactic acid, acetic acid, and succinic acid, and the overall production levels of acetic acid and succinic acid were increased by introducing the E. coli pntAB gene. Moreover, the NADH/NAD + ratio was increased by introduction of pntAB. Our results suggest that the E. coli PntAB transhydrogenase enhances the conversion of NADPH to NADH in C. glutamicum under microaerobic conditions, and the increased NADH/NAD + ratio results in increased succinic acid production. In addition, acetic acid production might be enhanced to supply ATP to the anaplerotic reaction catalyzed by pyruvate carboxylase.Key words: acetic acid; Corynebacterium glutamicum; pntAB; redox balance; succinic acid; transhydrogenases IntroductionMany oxidation and reduction reactions are involved in cellular metabolism, most of which require pyrimidine nucleotide cofactors, such as nicotinamide adenine dinucleotide (NAD + ) and nicotinamide adenine dinucleotide phosphate (NADP + ) and their reduced forms NADH and NADPH, to supply oxidizing and reducing equivalents. Generally, NADH is produced via catabolism, and is oxidized in the respiratory chain under aerobic conditions or by fermentation under anaerobic conditions. Anabolic reactions to produce cell biomass components, such as fatty acid and amino acid biosynthesis, require NADPH as a reducing equivalent. Therefore, maintaining the intracellular balance of NAD(P) + and NAD(P)H is very important for most intracellular metabolic reactions to occur.Some organisms contain the enzymes catalyzing the following interconversion of NADH and NADPH, called transhydrogenase:NADPH + NAD + NADP + + NADH. To date, two types of transhydrogenases have been identified; the cytoplasmic-soluble and membrane-bound types. Membrane-bound transhydrogenases have been found in the inner mitochondrial membrane of eukaryotic cells and in the cytoplasmic membrane of many bacteria, and simultaneously catalyze proton translocation across a membrane. Full PaperEnhanced acetic acid and succinic acid production under microaerobic conditions by Corynebacterium glutamicum harboring Escherichia coli transhydrogenase gene pntAB (Received April 3, 2014; Accepted April 8, 2014) Yuto Yamauchi, None of the authors of this manuscript ha...

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ACorynebacterium glutamicum rnhA recGDouble Mutant Showing Lysozyme- sensitivity, Temperature-sensitive Growth, and UV-Sensitivity

Cited by 13 publications

References 41 publications

The SCO2299 gene from Streptomyces coelicolor A3(2) encodes a bifunctional enzyme consisting of an RNase H domain and an acid phosphatase domain

The SCO2299 gene from Streptomyces coelicolor A3(2) encodes a bifunctional enzyme consisting of an RNase H domain and an acid phosphatase domain

Ribonuclease H: molecular diversities, substrate binding domains, and catalytic mechanism of the prokaryotic enzymes

Enhanced acetic acid and succinic acid production under microaerobic conditions by Corynebacterium glutamicum harboring Escherichia coli transhydrogenase gene pntAB

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