In this study, a combined whole-cell biotransformation process was used for efficient synthesis of optically pure (1R, 2S)-norephedrine [(1R, 2S)-NE]. The genes encoding R-selective acetohydroxyacid synthase I (AHAS I) from Escherichia coll and S-selective w-transaminase (omega-TA) from Chromobactertum violaceum BCRC10636 were cloned and over-expressed in E. colt NovaBlue cells. In the first biosynthetic step, L-phenylacetylcarbinol (L-PAC) was produced from benzaldehyde and pyruvate by using recombinant E. colt (pQE-AHAS I) cells, with almost 100% conversion yield and 71.8% purification yield. The purified L-PAC was coupled to L-alanine by using recombinant E. colt (pQE-CvTA) cells to produce (1R, 2S)-NE. This biocatalytic process was optimal at pH 6.5 to 8.0 and 37 degrees C, with a 1:10 ratio of L-PAC to L-alanine. Under the optimal conditions, the highest conversion yield of (1R, 2S)-NE was 62.2% and the enantiomeric excess value of (1R, 25)-NE was more than 99%. The recombinant E. coli (pQE-CvTA) cells could be reused for at least 20 cycles, with only a modest reduction in the conversion yield to 76.3% relative to the first cycle. Our results indicate that the combination of AHAS I- and omega-TA-expressing E. colt cells might be a potential biocatalyst for (1R, 2S)-NE production.