Synthesis of the alternative sweetener 5-ketofructose from sucrose by fructose dehydrogenase and invertase producing Gluconobacter strains

Hoffmann, Juliane; Hövels, Marcel; Kosciow, Konrad; Deppenmeier, Uwe

doi:10.1016/j.jbiotec.2019.11.001

Cited by 11 publications

(16 citation statements)

References 34 publications

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“…Additionally, the use of a promoter stronger than P GOX0264 could enhance fdhSCL expression and thereby 5-KF production. Besides fructose as substrate, 5-KF could also be produced from the cost-efficient and renewable feedstock sucrose when applying a sucrose-hydrolyzing enzyme and FDH (Hoffmann et al 2020). The fdhSCL genes were disruptively integrated into the mGDH gene locus under the control of the strong promoter P GOX0264 and the invertase Inv1417 gene was expressed under the control of the same promoter on a pBBR1MCS-2-based plasmid.…”

Section: Chromosomal Integrations and Intergenic Regions Suitable For Target Gene Expressionmentioning

confidence: 99%

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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Section: Chromosomal Integrations and Intergenic Regions Suitable For Target Gene Expressionmentioning

confidence: 99%

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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“…Gluconobacter oxydans has been widely applied in the industrial production of L-sorbose from D-sorbitol (De Wulf et al, 2000), dihydroxyacetone from glycerol (De La Morena et al, 2020), 1amino-L-sorbose from 1-amino-D-sorbitol (Schedel, 2000), and levan-type fructans from sucrose (Hövels et al, 2020). Furthermore, G. oxydans is also an excellent workhorse for the biosynthesis of 2-keto-D-gluconate (Li et al, 2016;Zhou et al, 2020), 5-keto-D-gluconate (Merfort et al, 2006), xylonic acid (Hahn et al, 2020;Shen et al, 2020), 5-keto-D-fructose (Battling et al, 2020;Hoffmann et al, 2020), and many other products (Deppenmeier et al, 2002;De Muynck et al, 2007). The wide applications of G. oxydans are mainly due to its unique dehydrogenases in the periplasm (Deppenmeier et al, 2002).…”

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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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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“…Therefore, these enzymes are also responsible for the production of some compounds such as fructooligosaccharides (FOSs), inulins, and levans [8,9]. To date, sucrolytic enzymes have diverse applications in the food [10][11][12], beverage [13,14], medicine [1,14], cosmetic [5,15], bioenergy [16], paper [15], and biochemical sectors [15]. Thus, the use of sucrolytic enzymes is highly widespread, with a large number of industrial applications.…”

Section: Introductionmentioning

confidence: 99%

Production of Sucrolytic Enzyme by Bacillus licheniformis by the Bioconversion of Pomelo Albedo as a Carbon Source

et al. 2021

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Recently, there has been increasing use of agro-byproducts in microbial fermentation to produce a variety of value-added products. In this study, among various kinds of agro-byproducts, pomelo albedo powder (PAP) was found to be the most effective carbon source for the production of sucrose hydrolyzing enzyme by Bacillus licheniformis TKU004. The optimal medium for sucrolytic enzyme production contained 2% PAP, 0.75% NH4NO3, 0.05% MgSO4, and 0.05% NaH2PO4 and the optimal culture conditions were pH 6.7, 35 °C, 150 rpm, and 24 h. Accordingly, the highest sucrolytic activity was 1.87 U/mL, 4.79-fold higher than that from standard conditions using sucrose as the carbon source. The purified sucrolytic enzyme (sleTKU004) is a 53 kDa monomeric protein and belongs to the glycoside hydrolase family 68. The optimum temperature and pH of sleTKU004 were 50 °C, and pH = 6, respectively. SleTKU004 could hydrolyze sucrose, raffinose, and stachyose by attacking the glycoside linkage between glucose and fructose molecules of the sucrose unit. The Km and Vmax of sleTKU004 were 1.16 M and 5.99 µmol/min, respectively. Finally, sleTKU004 showed strong sucrose tolerance and presented the highest hydrolytic activity at the sucrose concentration of 1.2 M–1.5 M.

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Synthesis of the alternative sweetener 5-ketofructose from sucrose by fructose dehydrogenase and invertase producing Gluconobacter strains

Cited by 11 publications

References 34 publications

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

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

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

Production of Sucrolytic Enzyme by Bacillus licheniformis by the Bioconversion of Pomelo Albedo as a Carbon Source

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