Surface display for metabolic engineering of industrially important acetic acid bacteria

Blank, Marshal; Schweiger, Paul

doi:10.7717/peerj.4626

Cited by 4 publications

(3 citation statements)

References 48 publications

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“…As mentioned previously, a few strong G. oxydans promoters have been reported (Hu et al, 2015;Li et al, 2016;Blank and Schweiger, 2018). To verify the strength of our strongest screened promoters, we also obtained four reported strong promoters (P tufB , P dnak , P hp0169 , and P 264 ) from the genomes of WSH-003 and ATCC 621H.…”

Section: Evaluation Of Promoter Strength By Measuring Mcherry Expressionmentioning

confidence: 72%

“…Though some progress were achieved in promoter discovery, it was hard to apply the promoters to other metabolic pathways because the promoters were relatively weak without systematic comparison. Saito et al, found some strong promoters of G. oxydans, such as P tufB , P 0169 , and P 264 (Saito et al, 1997;Yuan et al, 2016;Blank and Schweiger, 2018). Moreover, Kallnik and Hu reported some promoters of different strengths in G. oxydans (Kallnik et al, 2010;Hu et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Identification of Gradient Promoters of Gluconobacter oxydans and Their Applications in the Biosynthesis of 2-Keto-L-Gulonic Acid

Chen

Liu

et al. 2021

Front. Bioeng. Biotechnol.

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The acetic acid bacterium Gluconobacter oxydans is known for its unique incomplete oxidation and therefore widely applied in the industrial production of many compounds, e.g., 2-keto-L-gulonic acid (2-KLG), the direct precursor of vitamin C. However, few molecular tools are available for metabolically engineering G. oxydans, which greatly limit the strain development. Promoters are one of vital components to control and regulate gene expression at the transcriptional level for boosting production. In this study, the low activity of SDH was found to hamper the high yield of 2-KLG, and enhancing the expression of SDH was achieved by screening the suitable promoters based on RNA sequencing data. We obtained 97 promoters from G. oxydans’s genome, including two strong shuttle promoters and six strongest promoters. Among these promoters, P3022 and P0943 revealed strong activities in both Escherichia coli and G. oxydans, and the activity of the strongest promoter (P2703) was about threefold that of the other reported strong promoters of G. oxydans. These promoters were used to overexpress SDH in G. oxydans WSH-003. The titer of 2-KLG reached 3.7 g/L when SDH was under the control of strong promoters P2057 and P2703. This study obtained a series of gradient promoters, including two strong shuttle promoters, and expanded the toolbox of available promoters for the application in metabolic engineering of G. oxydans for high-value products.

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Section: Evaluation Of Promoter Strength By Measuring Mcherry Expressionmentioning

confidence: 72%

Section: Introductionmentioning

confidence: 99%

Identification of Gradient Promoters of Gluconobacter oxydans and Their Applications in the Biosynthesis of 2-Keto-L-Gulonic Acid

Chen

Liu

et al. 2021

Front. Bioeng. Biotechnol.

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“…In the present study, the CCD was employed to investigate the interactions between the significant variables and determine their optimum values for riboflavin production. A total of 30 experimental combinations for four significant variables (fructose, tryptone, K 2 HPO 4 , and CaCl 2 ) were achieved at five levels with the center points replicated six times in the experiment (run order: 4,18,20,22,23,29). The range of variables concentrations at different coded levels shown in Table 7.…”

Section: Genome Features and Identification Of Riboflavin In The G Omentioning

confidence: 99%

A novel strain of acetic acid bacteria Gluconobacter oxydans FBFS97 involved in riboflavin production

Noman

Al-Barha

Sharaf

et al. 2020

Sci Rep

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A novel bacterial strain of acetic acid bacteria capable of producing riboflavin was isolated from the soil sample collected in Wuhan, China. The isolated strain was identified as Gluconobacter oxydans FBFS97 based on several phenotype characteristics, biochemicals tests, and 16S rRNA gene sequence conducted. Furthermore, the complete genome sequencing of the isolated strain has showed that it contains a complete operon for the biosynthesis of riboflavin. In order to obtain the maximum concentration of riboflavin production, Gluconobacter oxydans FBFS97 was optimized in shake flask cultures through response surface methodology employing Plackett-Burman design (PBD), and Central composite design (CCD). The results of the pre-experiments displayed that fructose and tryptone were found to be the most suitable sources of carbon and nitrogen for riboflavin production. Then, PBD was conducted for initial screening of eleven minerals (FeSO 4 , FeCl 3 , KH 2 po 4 , K 2 HPO 4 , MgSO 4 , ZnSO 4 , NaCl, CaCl 2 , KCl, ZnCl 2 , and AlCl 3 .6H 2 O) for their significances on riboflavin production by Gluconobacter oxydans strain FBFS97. The most significant variables affecting on riboflavin production are K 2 HPO 4 and cacl 2 , the interaction affects and levels of these variables were optimized by CCD. After optimization of the medium compositions for riboflavin production were determined as follows: fructose 25 g/L, tryptone 12.5 g/L, K 2 HPO 4 9 g/L, and CaCl 2 0.06 g/L with maximum riboflavin production 23.24 mg/L. Riboflavin or the so-called vitamin B2 is a water-soluble vitamin that belongs to the B vitamins complex group, a vital component of the energy metabolism, and a bioactive molecule that has an important role in various cellular functions 1,2. In addition, to its role as the precursor of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN), the major products of riboflavin that act a fundamental role in metabolism, acting as cofactors for a wide variety of enzymes intermediating many redox reactions in the cell 3,4. In contrast, to many microorganisms and plants, humans and animals cannot synthesize this vitamin, and thus need to supply their diet with external riboflavin to meet their nutritional requirements 5. Riboflavin is used on a large scale as food and feed additives, food colorant, and pharmaceutical preparations. The commercial production of this vitamin can be accomplished by chemical synthesis or biological synthesis, yet in recent times the chemical synthesis has totally replaced to the microbial fermentation because of its cost-effectiveness, reduction in waste and energy requirements, and the use of renewable resources 6. At present, several species of bacteria and fungi are harnessed

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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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Surface display for metabolic engineering of industrially important acetic acid bacteria

Cited by 4 publications

References 48 publications

Identification of Gradient Promoters of Gluconobacter oxydans and Their Applications in the Biosynthesis of 2-Keto-L-Gulonic Acid

Identification of Gradient Promoters of Gluconobacter oxydans and Their Applications in the Biosynthesis of 2-Keto-L-Gulonic Acid

A novel strain of acetic acid bacteria Gluconobacter oxydans FBFS97 involved in riboflavin production

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

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