Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites

Table 3 Specific activities of AlsS, IlvC, IlvD, Kivd in cell extracts of Pseudomonas sp. strain VLB120, Pseudomonas sp. strain T7 pPAPC-Km, pPDPK and Ps. sp. strain VLB120 pCom10-kivd

	Specific activity (μmol min^-1mg_{total protein} ^-1)
	AlsS	IlvC	IlvD	Kivd
VLB120 wild type	0.033 ± 0.009	0.0257 ± 0.0017^*	0.0075 ± 0.0004	ND
VLB120 T7 pPAPC-Km, pPDPK	3.494 ± 0.299	0.1476 ± 0.0425^*	0.1955 ± 0.0023	2.96 ± 0.24
VLB120 pCOM10-kivd	n.d.	n.d.	n.d.	ND

ND: not detectable.
^*: coupled assay together with AlsS.
Cells were cultured in M9* pH 7.4, 20 g L^-1 glucose and. 5 g L^-1 yeast extract shake flasks at 30° Cells were induced for 4 h either with 0.05% DCPK (pCOM10) or with 1 mM IPTG (pPDPK, pPAPC-Km) in the early exponential phase. Errors indicate standard deviations (n = 2 - 3).

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