The biaxially oriented poly(lactic acid) films prepared using simultaneous biaxial stretching were perforated by CO2 laser with the power of 60 W. The focal spot diameter was fixed at 103 μm. Pulse durations were varied from 1 to 30 μs, which corresponds to the fluence from 1.4 to 42.6 J/cm2. The perforated microhole size increased with increasing laser energy. The shape of microholes was circular for the films of equi‐biaxial stretching, whereas elliptical microholes were formed with its long and short axes corresponding to the directions of higher and lower stretch ratios, respectively. Comparing the laser perforation behavior of the as‐drawn films and annealed films, the opening of a microhole in the annealed films started at higher laser fluence, and at the same fluence the size of microhole was smaller. Gas transmission rates of the biaxially oriented poly(lactic acid) films with different thicknesses were examined. The oxygen transmission rates (OTR) of film improved with increased microhole diameter. The OTR of films containing 1 microhole/103 cm2 (hole diameter of ~110 μm) was 184, 150, and 98 cm3/d in comparison with the OTR for the original films without microhole of only 16, 10, and 8 cm3/d, respectively. Packaging films with OTR higher than 100 cm3/d (equivalent to the 9700 cm3/m2/d) are required to create a modified atmosphere inside the package of fresh produce for shelf‐life extension. Measured OTR and CO2TR values of the 3 different shapes of microhole were compared and discussed.