beta-Galactosidases from Lactobacillus delbruekii subsp. bulgaricus 20056, Lb. casei 20094, Lactococcus lactis subsp. lactis 7962, Streptococcus thermophilus TS2, Pediococcus pentosaceus PE39 and Bifidobacterium bifidum 1901 were partially purified. The rate of hydrolysis of lactose given by the predominant beta-galactosidase activity from each of the bacteria studied was in all cases enhanced by Mg2+, while the effect of K+ and Na+ differed from strain to strain. The beta-galactosidases from all strains also catalysed trans-galactosylation reactions. The types of oligosaccharides produced appeared to be very similar in each case, but the rates of their production differed. All the beta-galactosidases were also capable of hydrolysing galactosyl-lactose although, unlike the other bacteria studied, Lb. delbruekii subsp. bulgaricus 20056 and Lc. lactis subsp. lactis 7962 were unable to utilise galactosyl-lactose as a carbon source for growth.
SummarySpectrophotometric assays ofβ–galactosidase (EC 3.2.1.23) and phospho-β–galactosidase (EC 3.2.1.85) activity were used to survey the lactose utilization pathways of lactic acid bacteria and bifidobacteria.β–Galactosidase activity was found in all six genera represented (Lactococcus, Streptococcus, Leuconostoc, Lactobacillus, PediococcusandBifidobacterium) while phospho-β–galactosidase was restricted to the lactococci, twoLactobacillusand twoLeuconostocspecies. A number of strains ofLactococcus lactis, Lactobacillus caseiandLeuconostocspp. contained both enzymes. Enzyme activities varied when cells were grown on different sugars, but in general were low or absent for cells grown on glucose compared with lactose. Two lactose-related compounds, lactulose and galactosyl lactose, believed to be specific growth factors for bifidobacteria, supported growth amongst a wide range of lactic acid bacteria in addition to bifidobacteria. Growth on galactosyl lactose was restricted to some but not all strains containingβ–galactosidase, implying that the presence ofβ–galactosidase is insufficient by itself to ensure utilization of galactosyl lactose. DNA fragments that encoded theLactococcus lactissubsp.cremorisphospho-β–galactosidase gene or theβ–galactosidase genes ofStreptococcus salivariussubsp.thermophilusorLactobacillus delbrueckiisubsp.bulgaricuswere isolated and used as probes in DNA-DNA hybridizations. Little or no hybridization was detected between these probes and plasmid or genomic DNA isolated from heterologous species, despite the presence of the corresponding enzyme activity in the strains probed.
Dairy process monitoring by application of multivariate curve resolution using alternating least squares is presented. Alternating least squares was used for resolving Fourier transform infrared spectral data from a dairy batch process in which lactose is enzymatically hydrolyzed to glucose and galactose. It was possible to extract four compounds (fat, lactose, and two other sugar components) from the spectral data obtained from nine process runs. Subsequently, the pure spectra obtained in this way were used to monitor the content of these compounds in two new process runs. In this way, alternating least squares made it possible to follow the hydrolysis process by Fourier transform infrared spectroscopy without the need for reference analyses. When the results were correlated to reference results for lactose, the accuracy was similar to that obtained when a partial least squares regression was performed on the same data; lactose correlation was 0.980 when alternating least squares was used and was 0.987 when partial least squares was used.
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