Improved succinate production by metabolic engineering.

Succinate is a promising chemical which has wide applications and can be produced by biological route. The history of the biosuccinate production shows that the joint effort of different metabolic engineering approaches brings successful results. In order to enhance the succinate production, multiple metabolical strategies have been sought. In this review, different overproducers for succinate production, including natural succinate overproducers and metabolic engineered overproducers, are examined and the metabolic engineering strategies and performances are discussed. Modification of the mechanism of substrate transportation, knocking-out genes responsible for by-products accumulation, overexpression of the genes directly involved in the pathway, and improvement of internal NADH and ATP formation are some of the strategies applied. Combination of the appropriate genes from homologous and heterologous hosts, extension of substrate, integrated production of succinate, and other high-value-added products are expected to bring a desired objective of producing succinate from renewable resources economically and efficiently.

Effects of pH and dissolved CO2 level on simultaneous production of 2,3-butanediol and succinic acid using Klebsiella pneumoniae.

The influences of pH and dissolved CO2 level on the regulation of growth and formation of catabolic end products have been investigated in Klebsiella pneumoniae. With increasing CO2 levels, there were no apparent changes in 2,3-butanediol production but succinic acid productions were enhanced significantly. A novel strategy for co-production of 2,3-butanediol and succinic acid using K. pneumoniae was developed by controlling pH and dissolved CO2 concentration in fermentation medium. Under the optimum condition, maximal 77.1 g l(-1) 2,3-butanediol and 28.7 g l(-1) succinic acid were obtained after 60 h of fed-batch fermentation, giving a 2,3-butanediol+succinic acid yield of 1.03 mol mol(-1) glucose. This type of fermentation producing two commercial interests at the same fermentation process might be considered for a promising biological production process which will decrease the production cost by sharing the operation and recovery cost.

Soluble expression of active human beta-defensin-3 in Escherichia coli and its effects on the growth of host cells.

BACKGROUND: Human beta-defensin-3 (HBD(3)) is an epithelial peptide that has been demonstrated to have a salt-insensitive broad spectrum of potent antimicrobial activity. Expressing antimicrobial peptides in Escherichia coli (E. coli) is very difficult for it can result in death of the bacterial host cells. Our aim was to establish a prokaryotic system expressing soluble HBD(3) protein and demonstrate the antimicrobial activity of the expressed protein. We then studied whether the host cells would activate the suicide pathways. METHODS: We first cloned the complementary DNA coding for the mature chain of HBD(3), inserted it into the vector PGEX-KG then transformed E. coli BL21 (DE3) with the appropriate recombinant plasmid. After induction with 0.5 mmol/L isopropyl-1-thio-beta-D-galactopyranoside (IPTG) the transformed E. coli produced a recombinant glutathione S-transferase and HBD(3) (GST-HBD(3)) fusion protein. The fusion protein was treated with thrombin to produce pure HBD(3) protein then the antimicrobial activity of HBD(3) was evaluated in a liquid microdilution assay. RESULTS: The fusion protein GST-HBD(3) was efficiently cleaved by thrombin and yielded HBD(3) that had anti-staphylococcus aureus activity with a minimal inhibitory concentration level of 12.5 microg/ml. The E. coli strain expressing the recombinant protein did not grow slower than the empty vector strain. CONCLUSION: Active HBD(3) in E. coli by expressing the recombinant protein GST-HBD(3) could be produced, and suicide did not occur in the E. coli strain expressing the recombinant protein.