Preparation of Organic Solventtolerant Mutants from Escherichia coli K-12.

Aono, Rikizo; Aibe, Kazuhiko; Inoue, Akira; Horikoshi, Koki

doi:10.1271/bbb1961.55.1935

Cited by 86 publications

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“…Previously, we have mapped one of the n-hexane resistance determinants at 1?2 min, between pdxA and araD, on the E. coli chromosome by genetic analysis of a n-hexanesensitive derivative, OST4251 (Aono et al, 1994b). This determinant was named ostA.…”

Section: Discussionmentioning

confidence: 99%

“…W2252thy is sensitive to n-hexane (log P ow 3?9) and resistant to diphenyl ether (log P ow 4?2). OST4251 is a JA300 derivative constructed by P1 transduction transferring the n-hexane-sensitive phenotype from W2252thy (Aono et al, 1994b). E. coli BL21(DE3) was used as the host strain to produce a truncated Imp protein.…”

Section: Methodsmentioning

confidence: 99%

“…chromosome by using P1 transduction analysis (Aono et al, 1994b) and observed that OST4251 acquired n-hexane resistance by introduction of pHB4251LN (Aono et al, 1994c). However, nucleotide sequence analysis showed that djlA might participate in n-hexane resistance.…”

Section: Assignment Of the Mutation Responsible For The N-hexane Sensmentioning

confidence: 99%

“…Previously, we have mapped a n-hexane resistance determinant at 1?2 min on the E. coli chromosome by genetic analyses of the n-hexane-resistant strain JA300 and its n-hexane-sensitive derivative (Aono et al, 1994b). At the time of that study, the gene encoding this resistance was designated ostA, denoting 'organic solvent tolerance'.…”

Section: Introductionmentioning

confidence: 99%

“…This was done for each bacterium on the basis of colony formation on LBGMg agar overlaid with an appropriate organic solvent (Aono et al, 1994b). The strength of the bactericidal effect of solvent is correlated inversely with the solvent log P ow value under the conditions tested.…”

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n-Hexane sensitivity of Escherichia coli due to low expression of imp/ostA encoding an 87 kDa minor protein associated with the outer membrane

et al. 2003

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Most Escherichia coli strains are resistant to n-hexane. E. coli OST4251 is a n-hexane-sensitive strain that was constructed by transferring the n-hexane-sensitive phenotype from a n-hexane-sensitive strain by P1 transduction. OST4251 is resistant to diphenyl ether, which is less harmful than n-hexane to micro-organisms. The genetic determinant responsible for this subtle difference in the solvent resistance is mapped at 1·2 min on the E. coli chromosome. Nucleotide sequence analysis showed that IS2 and IS5 had integrated upstream of the imp/ostA structural gene in OST4251. The integration of IS2 decreased the activity of the imp/ostA promoter. A product of the gene was identified immunologically as an 87 kDa minor protein associated with the outer membrane. Upon transformation with plasmids containing the imp/ostA gene, OST4251 produced a high level of the gene product in the membrane and acquired n-hexane resistance. Thus, the low level of promoter activity resulted in low Imp production and the n-hexane-sensitivity phenotype. It is likely that the gene product contributes to n-hexane resistance by reducing the influx of n-hexane.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Assignment Of the Mutation Responsible For The N-hexane Sensmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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n-Hexane sensitivity of Escherichia coli due to low expression of imp/ostA encoding an 87 kDa minor protein associated with the outer membrane

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Toxicity of Organic Solvents to Microorganisms

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2003

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Synthesis of 2,6‐Dihalogenated Purine Nucleosides by Thermostable Nucleoside Phosphorylases

et al. 2015

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The enzymatic transglycosylation of 2,6-dichloropurine (26DCP) and 6-chloro-2-fluoropurine (6C2FP) with uridine, thymidine and 1-(b-d-arabinofuranosyl)-uracil as the pentofuranose donors and recombinant thermostable nucleoside phosphorylases from G. thermoglucosidasius or T. thermophilus as biocatalysts was studied. Selection of 26DCP and 6C2FP as substrates is determined by their higher solubility in aqueous buffer solutions compared to most natural and modified purines and, furthermore, synthesized nucleosides are valuable precursors for the preparation of a large number of biologically important nucleosides. The substrate activity of 26DCP and 6C2FP in the synthesis of their ribo-and 2'-deoxyA C H T U N G T R E N N U N G ribo-nucleosides was closely similar to that of related 2-amino-(DAP), 2-chloro-and 2-fluoroadenines; the efficiency of the synthesis of b-d-arabinofuranosides of 26DCP and 6C2FP was lower vs. that of DAP under similar reaction conditions. For a convenient and easier recovery of the biocatalysts, the thermostable enzymes were immobilized on MagReSyn epoxide beads and the biocatalyst showed high catalytic efficiency in a number of reactions. As an example, 6-chloro-2-fluoro-(b-d-ribofuranosyl)-purine (9), a precursor of various antiviral and antitumour drugs, was synthesized by the immobilized enzymes at 60 8C under high substrate concentrations (uriA C H T U N G T R E N N U N G dine:purine ratio of 2:1, mol). The synthesis was successfully scaled-up [uridine (2.5 mmol), base (1.25 mmol); reaction mixture 50 mL] to afford 9 in 60% yield. The reaction reveals the great practical potential of this enzymatic method for the efficient production of modified purine nucleosides of pharmaceutical interest.

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Preparation of Organic Solventtolerant Mutants from Escherichia coli K-12.

Cited by 86 publications

References 0 publications

n-Hexane sensitivity of Escherichia coli due to low expression of imp/ostA encoding an 87 kDa minor protein associated with the outer membrane

n-Hexane sensitivity of Escherichia coli due to low expression of imp/ostA encoding an 87 kDa minor protein associated with the outer membrane

Toxicity of Organic Solvents to Microorganisms

Synthesis of 2,6‐Dihalogenated Purine Nucleosides by Thermostable Nucleoside Phosphorylases

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