Characterization of newly identified DnaA and DnaB proteins from Acetobacter

Bugala, Juraj; Cimová, Viera; Grones, Peter; Grones, Jozef

doi:10.1016/j.resmic.2016.06.010

Cited by 1 publication

(3 citation statements)

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“…Varying the length of the P lac region can increase or decrease target gene expression in Acetobacter likely due to 5'-UTR effects in the mRNA (Tonouchi et al 1998a). Only two studies reported to have used LacI-dependent IPTG-induced target gene expression (Bugala et al 2016;Liu et al 2020b). Only in one study the LacI-dependent IPTG-induced expression performance has been reported (Liu et al 2020b).…”

Section: Discussionmentioning

confidence: 99%

“…In one Acetobacter study, the IPTG-inducible P T7 -based Novagen vector pET-30a(+) with pBR322 origin was described to have been used in A. pasteurianus for inducible expression of the endogenous genes of DnaA-and DnaBlike proteins as well as the dnaA and dnaB genes from E. coli to study the replacement of the corresponding endogenous proteins expressed from the chromosome (Bugala et al 2016). A strong induction of the plasmid-based gene expression in A. orleanensis was reported, yet it was not explained whether it resulted from the T7 promoter present on pET-30a(+) or from another promoter included by the cloning procedure.…”

Section: Other Expression Plasmids Used In Acetobactermentioning

confidence: 99%

“…A strong induction of the plasmid-based gene expression in A. orleanensis was reported, yet it was not explained whether it resulted from the T7 promoter present on pET-30a(+) or from another promoter included by the cloning procedure. Furthermore, plasmid backbone pBAD18 carrying pBR322 origin, the classical L-arabinose-dependent AraC regulator gene araC and the P araBAD target promoter of AraC have been used in A. orleanensis for arabinose-inducible expression of asRNA fragments (121 nucleotides) which are antisense to the C-terminal coding sequence of the dnaA and dnaB gene sequences studied (Bugala et al 2016). The data of the asRNA induction by arabinose have not been shown, thus a conclusion about leakiness and induction performance is not possible yet.…”

Section: Other Expression Plasmids Used In Acetobactermentioning

confidence: 99%

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On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

Fricke

Klemm

Bott

et al. 2021

Appl Microbiol Biotechnol

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Acetic acid bacteria (AAB) are valuable biocatalysts for which there is growing interest in understanding their basics including physiology and biochemistry. This is accompanied by growing demands for metabolic engineering of AAB to take advantage of their properties and to improve their biomanufacturing efficiencies. Controlled expression of target genes is key to fundamental and applied microbiological research. In order to get an overview of expression systems and their applications in AAB, we carried out a comprehensive literature search using the Web of Science Core Collection database. The Acetobacteraceae family currently comprises 49 genera. We found overall 6097 publications related to one or more AAB genera since 1973, when the first successful recombinant DNA experiments in Escherichia coli have been published. The use of plasmids in AAB began in 1985 and till today was reported for only nine out of the 49 AAB genera currently described. We found at least five major expression plasmid lineages and a multitude of further expression plasmids, almost all enabling only constitutive target gene expression. Only recently, two regulatable expression systems became available for AAB, an N-acyl homoserine lactone (AHL)-inducible system for Komagataeibacter rhaeticus and an l-arabinose-inducible system for Gluconobacter oxydans. Thus, after 35 years of constitutive target gene expression in AAB, we now have the first regulatable expression systems for AAB in hand and further regulatable expression systems for AAB can be expected. Key points • Literature search revealed developments and usage of expression systems in AAB. • Only recently 2 regulatable plasmid systems became available for only 2 AAB genera. • Further regulatable expression systems for AAB are in sight.

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