Combining metabolic engineering and adaptive evolution to enhance the production of dihydroxyacetone from glycerol by Gluconobacter oxydans in a low-cost way

Lu, Leifang; Wei, Liujing; Zhu, Kun Yan; Wei, Dongzhi; Hua, Qiang

doi:10.1016/j.biortech.2012.03.013

Cited by 27 publications

(26 citation statements)

References 33 publications

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“…During the process, two hydrogen atoms were removed from one molecular of D-sorbitol to form one molecular of L-sorbose. Gluconobacter oxydans is widely used in biotransformation due to its ability to incompletely oxidize D-sorbitol, glycerol, and glucose to L-sorbose [ 5 ], dihydroxypropanone [ 6 ], and gluconic acid [ 7 ], respectively. These oxidation reactions are performed by membrane-bound dehydrogenases located on the outer surface of the cytoplasmic membrane, whereas the oxidation products accumulate in the culture medium.…”

Section: Introductionmentioning

confidence: 99%

“…The gene clusters encoding D-sorbitol dehydrogenase ( sldhAB , 2,670 bp) have been identified from the draft genome sequence of G. oxydans WSH-003 [ 8 ]. For industrial production, it is preferable to engineer G. oxydans cells to further enhance their catalytic properties during the biotransformation processes of D-sorbitol [ 6 , 10 , 11 ]. For example, by overexpressing a membrane-bound glycerol dehydrogenase gene in G. oxydans DSM 2343, the titer of dihydroxyacetone from glycerol was enhanced by 75% [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

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Enhanced production of L-sorbose from D-sorbitol by improving the mRNA abundance of sorbitol dehydrogenase in Gluconobacter oxydansWSH-003

Wang

et al. 2014

Microb Cell Fact

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BackgroundProduction of L-sorbose from D-sorbitol by Gluconobacter oxydans is the first step to produce L-ascorbic acid on industrial scale. The sldhAB gene, which encodes the sorbitol dehydrogenase (SLDH), was overexpressed in an industrial strain G. oxydans WSH-003 with a strong promoter, PtufB. To enhance the mRNA abundance, a series of artificial poly(A/T) tails were added to the 3′-terminal of sldhAB gene. Besides, their role in sldhAB overexpression and their subsequent effects on L-sorbose production were investigated.ResultsThe mRNA abundance of the sldhAB gene could be enhanced in G. oxydans by suitable poly(A/T) tails. By self-overexpressing the sldhAB gene in G. oxydans WSH-003 with an optimal poly(A/T) tail under the constitutive promoter PtufB, the titer and the productivity of L-sorbose were enhanced by 36.3% and 25.0%, respectively, in a 1-L fermenter. Immobilization of G. oxydans-sldhAB6 cells further improved the L-sorbose titer by 33.7% after 20 days of semi-continuous fed-batch fermentation.ConclusionsThe artificial poly(A/T) tails could significantly enhance the mRNA abundance of the sldhAB. Immobilized G. oxydans-sldhAB6 cells could further enlarge the positive effect caused by enhanced mRNA abundance of the sldhAB.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Enhanced production of L-sorbose from D-sorbitol by improving the mRNA abundance of sorbitol dehydrogenase in Gluconobacter oxydansWSH-003

Wang

et al. 2014

Microb Cell Fact

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“…In industrial production, it is preferable to obtain large amount of G. oxydans cells as biocatalyst on relatively inexpensive carbon sources. Previous studies indicated that the mutant strain deficient in mGDH could exhibit enhanced growth rate and biomass yield on glucose (Gupta et al 1997;Krajewski et al 2010;Lu et al 2012). In addition, the absence of mGDH eliminated the extracellular glucose oxidation and thus could facilitate the study of intracellular utilization of glucose.…”

Section: Introductionmentioning

confidence: 98%

Revealing in vivo glucose utilization of Gluconobacter oxydans 621H Δmgdh strain by mutagenesis

Wei

Zhu

Jing-lun

et al. 2014

Microbiological Research

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Gluconobacter oxydans, belonging to acetic acid bacteria, is widely used in industrial biotechnology. In our previous study, one of the main glucose metabolic pathways in G. oxydans 621H was blocked by the disruption of the mgdh gene, which is responsible for glucose oxidation to gluconate on cell membrane. The resulting 621H Δmgdh mutant strain showed an enhanced growth and biomass yield on glucose. In order to further understand the intracellular utilization of glucose by 621H Δmgdh, the functions of four fundamental genes, namely glucokinase-encoding glk1 gene, soluble glucose dehydrogenase-encoding sgdh gene, galactose-proton symporter-encoding galp1 and galp2 genes, were investigated. The obtained metabolic characteristics of 621H Δmgdh Δglk1 and 621H Δmgdh Δsgdh double-gene knockout mutants showed that, in vivo, glucose is preferentially phosphorylated to glucose-6-phosphate by glucokinase rather than being oxidized to gluconate by soluble glucose dehydrogenase. In addition, although the galactose-proton symporter-encoding genes were proved to be glucose transporter genes in other organisms, both galp genes (galp 1 and galp2) in G. oxydans were not found to be involved in glucose uptake system, implying that other unknown transporters might be responsible for transporting glucose into the cells.

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“…Microorganisms have ability to adapt themselves rapidly to different environments. This property has been gradually developed and utilized in varied microorganisms by conducting adaptive laboratory evolution (ALE) experiments to improve the activity or tolerance of strains (Lu et al, 2012). The selective pressure is very important to the success of ALE, and the pressures can be divided in two main categories: nutrient stress and environmental stress (Nam et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Adaptive evolution of Schizochytrium sp. by continuous high oxygen stimulations to enhance docosahexaenoic acid synthesis

Sun

Ren

et al. 2016

Bioresource Technology

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Adaptive laboratory evolution (ALE) is an effective method in changing the strain characteristics. Here, ALE with high oxygen as a selection pressure was applied to improve the production capacity of Schizochytrium sp. Results showed that cell dry weight (CDW) of endpoint strain was 32.4% higher than that of starting strain. But slight lipid accumulation impairment was observed. These major performance changes were accompanied with enhanced isocitrate dehydrogenase enzyme activity and reduced ATP:citrate lyase enzyme activity. And a serious decrease of 62.6% in SDHA 140rpm→170rpm was observed in the endpoint strain. To further study the docosahexaenoic acid (DHA) production ability of evolved strain, fed-batch strategy was applied and 84.34g/L of cell dry weight and 26.40g/L of DHA yield were observed. In addition, endpoint strain produced greatly less squalene than starting strain. This work demonstrated that ALE may be a promising tool in modifying microalga strains.

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Combining metabolic engineering and adaptive evolution to enhance the production of dihydroxyacetone from glycerol by Gluconobacter oxydans in a low-cost way

Cited by 27 publications

References 33 publications

Enhanced production of L-sorbose from D-sorbitol by improving the mRNA abundance of sorbitol dehydrogenase in Gluconobacter oxydansWSH-003

Enhanced production of L-sorbose from D-sorbitol by improving the mRNA abundance of sorbitol dehydrogenase in Gluconobacter oxydansWSH-003

Revealing in vivo glucose utilization of Gluconobacter oxydans 621H Δmgdh strain by mutagenesis

Adaptive evolution of Schizochytrium sp. by continuous high oxygen stimulations to enhance docosahexaenoic acid synthesis

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