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a b s t r a c tTwo commercial available lactases from Aspergillus oryzae and Kluyveromyces lactis were used to study the synthesis of galactooligosaccharides (GOS) in sweet and acid whey. At 38 g L À1 initial lactose concentration, the A. oryzae enzyme gave a GOS yield of 10.91 ± 0.01% in lactose solution, 10.93 ± 0.18% in sweet whey and 11.32 ± 0.59% in acid whey. Thus, the components in whey did not influence the enzymes transgalactolytic activity. On the other hand, the K. lactis enzyme showed a strong dependence on whey type and whey concentration. At 38 g L À1 initial lactose concentration, GOS yields were 10.93 ± 0.26% in lactose solution, 4.30 ± 0.17% in sweet whey and 10.56 ± 0.41% in acid whey. However, with increasing initial lactose concentration, the inhibitory effect of sweet whey was decreasing, which resulted in even higher yields than in lactose solution.

Galactooligosaccharides (GOS) are synthesized by the enzyme β-galactosidase during the hydrolysis of lactose. In this so-called transgalactosylation reaction the galactosyl moiety is transferred to another sugar molecule instead of water resulting in oligosaccharides of different chain lengths and glycosidic linkages. Because their structures are similar to oligosaccharides present in human breast milk, they act as prebiotics, which has been shown for infants and adults to be alike. While so far most of the research to maximize GOS yield has been carried out using buffered lactose solution as a starting material, more and more work is now conducted with dairy by-products such as whey and whey permeate, or even milk, for direct GOS synthesis in order to develop new GOS-enriched dairy products. This review aims to summarize the results obtained with various dairy liquids, and it rates their suitabilities to act as raw material for GOS production. Most of the studies using whey or milk have been carried out with enzymes from Aspergillus oryzae, Kluyveromyces lactis, Bacillus circulans, Streptococcus thermophilus, and several Lactobacillus species. As the initial lactose concentration (ILC) is known to be a crucial factor for high GOS yield, most of the research has been done with concentrated or supplemented milk and whey. However, a clear dependency on ILC could only be observed for the A. oryzae lactase, indicating a strong influence of milk components like minerals and proteins on the transfer activities of most enzymes.

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BackgroundThe enzyme cellobiose dehydrogenase (CDH) can be used to oxidize lactose to lactobionic acid. As Sclerotium rolfsii is known to be a good producer of CDH, the aim of this paper was to simplify its production and secondly to systematically study its purification aiming for a high yield. Two preservation methods (freezing and freeze-drying) and the influence of several protectants were investigated.ResultsProduction of cellobiose dehydrogenase was optimized leading to a more simplified medium composition. Purification of the enzyme was evaluated by determining breakthrough profiles on different ion exchange (IEX) and hydrophobic interaction (HIC) materials with regard to buffer composition. Highest purification with an acceptable loss during the capture step using IEX was obtained with a Q Sepharose XL medium and a 100 mM sodium acetate buffer at pH 4.5. Subsequent purification using hydrophobic interaction chromatography was done at 1.1 M ammonium sulfate concentration. Purification was moderate, yielding a specific activity of 11.9 U/mg (56% yield). However, as could be shown in a preliminary experiment, purity of the obtained enzyme solution was sufficient for its intended use to oxidize lactose to lactobionic acid. Various sugars and sugar alcohols were investigated to study their protective effect during lyophilisation and freezing at -20°C. Glucose and lactulose could be identified to have a high lyoprotective effect while loss of enzyme activity was high (77%) when using no additives.ConclusionBy simplifying the cultivation medium of Sclerotium rolfsii, the costs of cellobiose dehydrogenase production could be reduced. Simultaneously, CDH production was increased by 21%. The production of lactobionic acid from lactose is possible using partially purified and unpurified enzyme. Storage at -20°C using 50% (w/v) glycerol was considered to be most suited for preservation of the enzyme.