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Sophorolipids, obtained by a two-stage process starting from deproteinized whey concentrate using Cryptococcus curvatus ATCC 20509 and Candida bombicola ATCC 22214, were compared to products from one-stage processes, using different lipidic compounds as substrates. Results showed that above all carbon source and not cultivation conditions had a distinct influence on the composition of the crude product mixture and therefore on the physicochemical and biological properties of the sophorolipids, such as, for example, surface activity, cytotoxicity and stability against hydrolases. The results were completed by corresponding data for purified mono- and diacetylated (17-hydroxyoctadecenoic)-1′,4′′-lactonized sophorolipids. Crude sophorolipid mixtures showed moderate to good surface active properties (SFTmin 39 mN m−1, CMC 130 mg l−1), water solubilities (2–3 g l−1) and low cytotoxicities (LC50 300–700 mg l−1). In contrast, purified sophorolipids were more surface active (SFTmin 36 mN m−1, CMC 10 mg l−1), less water soluble (max. 70 mg l−1) and showed stronger cytotoxic effects (LC50 15 mg l−1). Incubation of crude sophorolipid mixtures with different hydrolases demonstrated that treatment with commercially available lipases such as from Candida rugosa and Mucor miehei distinctly reduced the surface active properties of the sophorolipids, while treatment with porcine liver esterase and glycosidases had no effect.

In order to produce sophorolipids from whey, thereby lowering the lactose content and biological oxygen demand, a two-step batch cultivation process was developed including medium sterilization by filtration. In the first step, whey was sterilized by a combination of crossflow and sterile filtration. Because the sophorolipid-producing yeast Candida bombicola ATCC 22214 was not able to use lactose as a carbon source directly, the oleaginous yeast Cryptococcus curvatus ATCC 20509 was grown on deproteinized whey concentrates (DWC). With 1:1 diluted DWC-20, lactose was consumed as the carbon source and biomass (24 g/l dry weight content) as well as single-cell oil (SCO, 10 g/l) were produced. The cultivation broth was disrupted with a glass bead mill and it served as medium for growth (29 g cell dry mass/l) and sophorolipid production (12 g/l) of the yeast C. bombicola.

High concentrations of 422 g sophorolipids l−1 were produced using a two-stage cultivation process: first deproteinized whey concentrate (DWC) containing 110 g lactose l−1 was used for cultivation of the yeast Cryptococcus curvatus ATCC 20509, resulting in 34 g dry weight l−1, 20 g single-cell oil l−1 and reducing the chemical oxygen demand (COD) from 159 g l−1 to 35 g oxygen l−1. Afterwards cells were disrupted by passing the cell suspension directly through a high pressure laboratory homogeniser. After autoclavation, the resulting crude cell extract containing the single-cell oil served as substrate for growth of Candida bombicola ATCC 22214 and for sophorolipid production in a second stage. When the single-cell oil was consumed, repeated feeding of 400 g rapeseed oil l−1 was started increasing the yield of sophorolipids to 422 g l−1. A simple technique for product isolation, sedimentation, could be used to harvest the crude sophorolipids. © Rapid Science Ltd. 1998