RÉSUMÉ
In this study, a newly strain named Clostridium butyricum YJH-09 were isolated from the sample of pond soil and identified through physiological, biochemical and 16S rDNA analysis. Then, the dhaT gene encoding a novel 1,3-propanediol dehydrogenase (PDOR) was cloned from this strain and expressed in Escherichia coli BL21(DE3). Subsequently, the recombinant PDOR was purified and the optimal pH and temperature, specific activities and kinetic parameter were investigated. Furthermore, the whole cells of Clostridium butyricum YJH-09 mixed with BL21-dhaT were used to produce 1,3-PD through co-biotransformation. As results, 25.88g/L of 1,3-PD was generated with 0.54g/g yield from 50g/L glycerol in 30h, and the 1,3-PD production was increased more than 2-fold compared with wild type strain alone. This research would offer useful information for further development of the biosynthesis of 1,3-PD.
Sujet(s)
Alcohol dehydrogenase , Clostridium butyricum/métabolisme , Propylène glycols , Biotransformation , ADN bactérien , Fermentation , GlycérolRÉSUMÉ
In the present study, a new strain of Lactobacillus brevis producing d-tagatose was isolated and identified. Then, the l-arabinose isomerase (L-AI) of this strain was displayed on the spore surface of Bacillus subtilis DB403 by using an anchoring protein CotG and a peptide linker (Gly-Gly-Gly-Gly-Ser). This displayed L-AI with high specific activity and stability was used as a novel immobilized biocatalyst for producing d-tagatose through batch and semi-continuous biotransformation. The conversion rate of d-tagatose from 125â¯g/L d-galactose was achieved 79.7% at 28â¯h, and the volumetric productivity reached 4.3â¯g/L/h at 20â¯h. Furthermore, the displayed L-AI showed a good performance on the reusability and remained 87% of the specific activity and 40.7% of the conversion rate after five recycles. A high efficient immobilized method for producing food-grade d-tagatose was established using spore surface-displayed L-AI.
Sujet(s)
Aldose-ketose isomerases/métabolisme , Hexose/métabolisme , Levilactobacillus brevis/enzymologie , 4-Chloro-7-nitro-2,1,3-benzoxadiazole/analogues et dérivés , Biotransformation , Galactose , Lysine/analogues et dérivés , Spores bactériensRÉSUMÉ
1,3-Propanediol (1,3-PD) is one of the most important chemicals widely used as monomers for synthesis of some commercially valuable products, including cosmetics, foods, lubricants and medicines. Although 1,3-PD can be synthesized both chemically and biosynthetically, the latter offers more merits over chemical approach as it is economically viable, environmentally friendly and easy to carry out. The biosynthesis of 1,3-PD can be done by transforming glycerol or other similar substrates using some bacteria, such as Clostridium butyricum and Klebsiella pneumoniae. However, these natural microorganisms pose some bottlenecks like low productivity and metabolite inhibition. To overcome these problems, recent research efforts have been focused more on the development of new strains by modifying the genome through different techniques, such as mutagenesis and genetic engineering. Genetically engineered strains obtained by various strategies cannot only gain higher yield than wild types, but also overcome some of the barriers in production by the latter. This review paper presents an overview on the recent advances in the technological approaches to develop genetically engineered microorganisms for efficient biosynthesis of 1,3-PD.