A bacterium strain B26 capable of producing (R)-alpha-hydroxyphenylacetic acid [(R)-HPA] (yield, 47.5%; enantiomeric excess, 99.1%) from phenylglyoxylic acid (PGA) with high optical purity was isolated and identified as Bacillus sp. B26 by 16S rDNA (ribosomal DNA) sequencing. Phylogenic analysis showed that the strain was most similar to Bacillus sp. enrichment culture clone SYW5 (FJ601635.1) and Bacillus cereus strain FM-4 (EU794727.1). Efforts were made to further improve HPA-production and PGA-tolerance by UV irradiation and UV-LiCl cooperative mutagenesis. Among viable mutants, B. sp. UV-38 and B. sp. ULi-11 exhibited better productivities than the wild type. Comparisons of HPA production and time course among wild strain and two mutants showed that B. sp. ULi-11 was more competent than B. sp. UV-38. HPA production was increased by 39.1% with B. sp. ULi-11 (yield, 65.4%) compared to that with B. sp. B26 (yield, 47.0%) when cultured in fermentation broth (pH 7.2) at 32 degrees C with an agitation speed of 180 r.min(-1) and PGA final concentration of 15 mmol.L-1 for 25 h.