Production of Lactase by Candida pseudotropicalis Grown in Whey

Lactase (β-d-galactosidase) was produced by Candida pseudotropicalis grown in deproteinized whey. Maximum enzyme production in 2% whey was obtained by supplementation with 0.15% yeast extract, 0.1% (NH4)2SO4, and 0.05% KH2PO4 (wt/vol). Highest enzyme values (4.35 U/mg of cells and 68 U/ml) were obtained with 10 to 12% whey, while enzyme yield was maximal in 2% whey (0.87 U/mg of whey). Optimal initial pH for cultivation was 3.5. The best conditions for extraction included 2% (wt/vol) chloroform, 10 h of treatment, pH 6.6 and higher, and 30 to 37°C. Optimum pH and temperature for enzyme activity were 6.2 and 47°C. The enzyme had a Km for O-nitrophenyl-β-d-galactopyranoside of 3.06 × 10−3 M and the initial Vmax was estimated as 6.63 × 10−8 M per min. It hydrolized 50 and 100% of the lactose in whey and milk within 4 and 5 h, respectively, at 37°C. The lyophilized enzyme retained 95% of activity for 3 months when stored at −20°C.

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