FIG. 1.

Construction of the ACC deaminase-producing A. tumefaciens strain. (A) Plasmid construction for expression of ACC deaminase in A. tumefaciens. An HindIII and XbaI fragment (1 kb) containing the ACC deaminase gene from Pseudomonas sp. strain ACP was ligated into the HindIII and XbaI sites of the broad-host-range plasmid pBBR1MCS-5, resulting in pBBRacdS. The expression of the ACC deaminase gene acdS was under the control of the lac promoter. MCS stands for multiple cloning sites. (B) Detection of ACC deaminase activity in A. tumefaciens. The α-ketobutirate (α-KB) accumulation in the reaction buffer was measured according to the method of Honma and Shimomura (7). The triangles and circles indicate the lysates from A. tumefaciens C58C1Rif^R(pBBR1MCS-5, pIG121-Hm) and C58C1Rif^R(pBBRacdS, pIG121-Hm), respectively. Closed and open symbols represent samples with and without ACC in the reaction buffer, respectively. Bars indicate standard deviations (n = 3).