Uridine phosphorylase from Shewanella oneidensis MR-1: Heterological expression, regulation, transcription, and properties

Mordkovich, N. N.; Manuvera, Valentin A.; Вейко, В. П.; Дебабов, В. Г.

doi:10.1134/s0003683812090037

Cited by 8 publications

(11 citation statements)

References 16 publications

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“…It is possible to assume that a decrease in the con tent of recombinant SEB, SEH, and SAV proteins in the periplasmic fraction of S. oneidensis MR 1 trans formants is a consequence of both an insufficiently successful expression of the studied genes in the S. oneidensis MR 1 cells under the control of selected promoter-operator region (although it contradicts previously obtained data [12,35]) and a decrease in the efficiency of translocation of the recombinant pro proteins in the periplasmic space. Analysis of the proteins of cytoplasmic fraction of the S. oneidensis MR 1/pLSEB/pLSEH/pLSAV/pRHKm transfor mant strains by the method of polyacrylamide gel elec trophoresis under denaturing conditions (Fig.…”

Section: Resultsmentioning

confidence: 57%

“…The plasmids containing pro enterotoxins B, H (SEB, SRH) and pro streptavidin (SAV) genes were previously constructed [7,9,18,19]. The pER plasmid containing the kanamycin resis tance gene and promoter-operator region of the udp gene from E. coli was previously constructed [12]. The pACYC177 plasmid was obtained from RNCIM of the Research Institute for Genetics and Selection of Industrial Microorganisms.…”

Section: Methodsmentioning

confidence: 99%

“…At present, the Shewanella oneidensis MR 1 pro teobacteria are widely used as a recipient strain for the homo and heterologous expression of different genes, by which it is possible to obtain post translationally modified recombinant proteins, including those for medical purposes (particularly, cytochromes), and to determine their structural and functional organization [10][11][12][13].…”

Section: Introductionmentioning

confidence: 99%

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Investigation of protein translocation Sec-system with heterologous gene expression in Shewanella oneidensis MR-1 bacterium cells

Mordkovich

Okorokova

Вейко

2015

Appl Biochem Microbiol

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A comparison of the primary structures of the protein translocation Sec system proteins in the Shewanella oneidensis MR 1 and Escherichia coli bacteria was carried out. The process of translocation of recombinant pro enterotoxins (SEB and SEH) from Staphylococcus aureus and pro streptavidin (SAV) from Streptomyces avidinii in the S. oneidensis MR 1 and E. coli cell periplasm was studied. It was demonstrated that these marker proteins are transferred into the periplasmic space of the S. oneidensis MR 1 transformant strain cells. The identity of N terminal amino acid sequences of mature recombinant SEB, SEH, and SAV proteins (generated during post translation proteolysis of leader peptide by the Sec system both in E. coli and S. oneidensis MR 1) was established.

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Section: Resultsmentioning

confidence: 57%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Investigation of protein translocation Sec-system with heterologous gene expression in Shewanella oneidensis MR-1 bacterium cells

Mordkovich

Okorokova

Вейко

2015

Appl Biochem Microbiol

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“…There are very few reports referring to recombinant expression in Shewanella currently. The heterologous expression of the udp genes from S. oneidensis MR-1 has been reported (Mordkovich et al 2012(Mordkovich et al , 2013. Besides, the promoter, tac, coupled with a synthetic flavin biosynthesis pathway from Bacillus subtilis has been shown to enhance extracellular electron-transfer (EET) rate of S. oneidensis MR-1 (Yang et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Turn on the Mtr pathway genes under pLacI promoter in Shewanella oneidensis MR-1

Guo

Zhou

et al. 2018

Bioresour. Bioprocess.

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Background: Shewanella genus is famous for applications like electron transfer in microbe fuel cells and bioremediation of heavy metals through the Mtr pathway. A potential way to enhance the electron genesis ability of Shewanella is to express exogenous mtr genes via recombinant DNA technology. Thus, to design and develop expression vectors capable of replicating in Shewanella and enhance the genetic toolbox of the same is important. Result: In this study, a plasmid construct with a replication origin, repB, and pLacI promoter is reported for the first time to drive the expression of green fluorescent protein in S. oneidensis MR-1. Based on the same vector, the Mtr pathway genes mtrA, mtrC, and mtrCAB were also successfully expressed in MR-1. The recombinant strains had higher ferric reductase activity compared to the wild type. The highest enzymatic activity of 508.33 U/L in genetic Shewanella with mtrC gene is obtained, which is 1.53-fold higher than that of wild strain. The plasmids were stable up to 90 generations. Conclusion: We have demonstrated an expression system based on pLacI promoter and repB ori in Shewanella. Consequently, the combination of repB and pLacI will have great potential in Shewanella to turn on expression of different genes constitutively.

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“…The gene expression of S. oneidensis MR 1 udp (GenBank GQ294526) was performed in the recipient strain E. coli, and the protein was isolated and purified as was described in [10]. The monomer subunit of SUP consists of 252 amino acid residues and has a molecular weight of 27 kDa.…”

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confidence: 99%

Physicochemical characterization of uridine phosphorylase from Shewanella oneidensis MR-1

Mordkovich

Safonova

Manuvera

et al. 2013

Dokl Biochem Biophys

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Uridine phosphorylase from Shewanella oneidensis MR-1: Heterological expression, regulation, transcription, and properties

Cited by 8 publications

References 16 publications

Investigation of protein translocation Sec-system with heterologous gene expression in Shewanella oneidensis MR-1 bacterium cells

Investigation of protein translocation Sec-system with heterologous gene expression in Shewanella oneidensis MR-1 bacterium cells

Turn on the Mtr pathway genes under pLacI promoter in Shewanella oneidensis MR-1

Physicochemical characterization of uridine phosphorylase from Shewanella oneidensis MR-1

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