Biosynthesis of poly(3-hydroxypropionate) from glycerol using engineeredKlebsiella pneumoniaestrain without vitamin B₁₂

Abstract

(2015) Biosynthesis of poly(3-hydroxypropionate) from glycerol using engineered Klebsiellapneumoniae strain without vitamin B 12 , Bioengineered, 6:2, 77-81, DOI: 10.1080/21655979.2015 The present study examined the production of P3HP from glycerol in the recombinant Klebsiella pneumoniae strain, which naturally synthesizes vitamin B 12 . The genes glycerol dehydratase and its reactivation factor (dhaB123, gdrA, and gdrB from K. pneumoniae), aldehyde dehydrogenase (aldH from E. coli) were cloned and expressed in K. pneumoniae to produce 3-hydroxypropionate (3HP), with 2 genes (dhaT and yqhD) for biosynthesis of 1,3-propanediol were deleted. To obtain P3HP production, propionyl-CoA synthetase (prpE from E. coli) and polyhydroxyalkanoate synthase (phaC from Ralstonia eutropha) were introduced. Under the appropriate aeration condition, the cell yield and P3HP content were 0.24 g/L and 12.7% (wt/wt [cell dry weight]) respectively along with 2.03 g/L 3HP after 48 h cultivation. Although the yield is relatively low, this study shows the feasibility of producing P3HP in K. pneumoniae from glycerol without vitamin B 12 for the first time. The results also suggest that the aeration conditions should be optimized carefully for the efficient production of P3HP.

Poly(3-hydroxypropionate): Biosynthesis Pathways and Malonyl-CoA Biosensor Material Properties

“…β-alanine route (– – –): poly(3HP) synthesis from glucose and glycerol. Aspartate biosynthesis, PanD, PP0596, YdfG, PrpE, PhaC1 ( Steinbüchel and Lütke-Eversloh, 2003 ; Andreeßen et al, 2010 , 2014b ; Wang et al, 2012 , 2013 ; Feng et al, 2015 ; Lacmata et al, 2017 ).…”

“…The introduction of the propionyl-CoA synthetase gene prpEEc of E. coli into the K. pneumonia strain ( Figure 1 ) resulted in aeration conditions that play a crucial role in poly(3HP) production and cell growth. Among the engineered metabolic pathways studied by Feng et al (2015) , 3-HP was used as a poly(3HP) precursor. However, poly(3HP) production accounted for only 10% of 3-HP production under three aeration conditions with different shaking culture rates of 50, 100, and 200 r/min ( Feng et al, 2015 ).…”

“…Among the engineered metabolic pathways studied by Feng et al (2015) , 3-HP was used as a poly(3HP) precursor. However, poly(3HP) production accounted for only 10% of 3-HP production under three aeration conditions with different shaking culture rates of 50, 100, and 200 r/min ( Feng et al, 2015 ).…”

Enhanced poly(3-hydroxypropionate) production via β-alanine pathway in recombinant Escherichia coli

“…The P3HP was extracted from the lyophilized cells with hot chloroform in a soxhlet apparatus and precipitated by the ice cold absolute ethanol as described [ 7 ]. The structure of the obtained P3HP was confirmed by NMR analysis using the Advanced III 600 NMR spectrometer (Bruker, Switzerland) as described in our previous study [ 9 ]. P3HP content was calculated using the ratio of P3HP weight to cell dry weight.…”

A novel autolysis system controlled by magnesium and its application to poly (3-hydroxypropionate) production in engineered Escherichia coli

“…The microbial production of P3HP has been achieved by genetically engineered strains so far. 8,9 However, few efforts have been devoted to the application of the autolysis system to P3HP fermentation. To overcome the present problems, we designed and constructed a new autolysis system by introducing the mgtA-UTR as a Mg 2C -responsive riboswitch.…”