Relative effectiveness of yeast cell wall digesting enzymes on Yarrowia lipolytica

Novozym 234 at a concentration of 1.0 mg/ml yielded 95.5% spheroplasts within 30 min at 37 degrees C, pH 7.0, with 36% regeneration which was the highest level of regeneration observed. Yeast lytic enzyme at a concentration of 1.0 mg/ml yielded 99.8% spheroplasts with only 1.5% regeneration. Glusulase was significantly less active in producing osmotically sensitive cells. All three enzymes yielded significantly higher levels of osmotically sensitive cells when cells were harvested from the mid-logarithmic phase of growth compared with later growth phases. beta-Glucuronidase failed to produce osmotically sensitive cells regardless of the phase of growth from which cells were harvested.