• Korean Journal of Food Science and Technology
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• v.22 no.4
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• pp.426-433
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• 1990

The conditions for immobilization of the partially purified ${\beta}-galactosidase$ form Bacillus subtilis HP4 and the properties of the immobilized enzyme have been investigated. The crude enzyme precipitated with cold acetone was purified about 68-fold through DEAE-cellulose and sephadex G-100 chromatography and its recovery was 19.9% The optimal conditions for Immobilization of enzyme were obtained in 2%(w/v) sodium alginate, 15%(v/v) enzyme solution and 2%(w/v) calcium chloride, and also the optimal stirring thme was 2 hours on the above conditions. The optimum temperature and pH values for immobilized enzyme were $55^{\circ}C$ and 6.5, respectively. Its residual activity was show 25% after heat treatment for an hour at $65^{\circ}C$, and found its high stability in pH 6.0 to 8.0. The enzyme activity was not affected b)· EDTA, 2-mercaptoethanol, KCN, protective agents, and other methal ions except Hg ion and Cu ion. The $K_m\;and\;V_{max}$ values of the immobilized enzyme on ONPG were $1.82{\times}10^{-2}M\;and\;3.57{\times}10^{-8}mole/min$, whereas those on lactose were $2.94{\times}10^{-2}M\;and\;1.68{\times}10^{-7} mole/min$, respectively. The remained enzyme activity for the immobilized enzyme was 95%t of original activity after storage of 40 days at $4^{\circ}C$, and when reused for 5 times was 81%. When skim milk(4.8% lactose) and 5% lactose solution were reacted with the immobilized enzyme(250 units/g) of lactose were 51% and 43%, respectively.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.454-460
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• 2007

Enzymatic properties and substrate specificity of recombinant $\beta-glycosidases$ from a hyperthermophilic archaeon, Sulfolobus shibatae (rSSG), were analyzed. rSSG showed its optimum temperature and pH at $95^{\circ}C$ and pH 5.0, respectively. Thermal inactivation of rSSG showed that its half-life of enzymatic activity at $75^{\circ}C$ was 15 h whereas it drastically decreased to 3.9 min at $95^{\circ}C$. The addition of 10 mM of $MnCl_2$ enhanced the hydrolysis activity of rSSG up to 23% whereas most metal ions did not show any considerable effect. Dithiothreitol (DTT) and 2-mercaptoethanol exhibited significant influence on the increase of the hydrolysis activity of rSSG rSSG apparently preferred laminaribiose $(\beta1\rightarrow3Glc)$, followed by sophorose $(\beta1\rightarrow2Glc)$, gentiobiose $(\beta1\rightarrow6Glc)$, and cellobiose $(\beta1\rightarrow4Glc)$. Various. intermolecular transfer products were formed by rSSG in the lactose reaction, indicating that rSSG prefers lactose as a good acceptor as well as a donor. The strong intermolecular transglycosylation activity of rSSG can be applied in making functional oligosaccharides.

• Journal of Microbiology
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• v.44 no.4
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• pp.396-402
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• 2006

[ ${\beta}-Galactosidase$ ] is extensively employed in the manufacture of dairy products, including lactose-reduced milk. Here, we have isolated two gram-negative and rod-shaped coldadapted bacteria, BS 1 and HS 39. These strains were able to break down lactose at low temperatures. Although two isolates were found to grow well at $10^{\circ}C$, the BS 1 strain was unable to grow at $37^{\circ}C$. Another strain, HS-39, evidenced retarded growth at $37^{\circ}C$. The biochemical characteristics and the results of 16S rDNA sequencing identified the BS 1 isolate as Rahnella aquatilis, and showed that the HS 39 strain belonged to genus Buttiauxella. Whereas the R. aquatilis BS 1 strain generated maximal quantities of ${\beta}-galactosidase$ when incubated for 60h at $10^{\circ}C$, Buttiauxella sp. HS-39 generated ${\beta}-galactosidase$ earlier, and at slightly lower levels, than R. aquatilis BS 1. The optimum temperature for ${\beta}-galactosidase$ was $30^{\circ}C$ for R. aquatilis BS-1, and was $45^{\circ}C$ for Buttiauxella sp. HS-39, thereby indicating that R. aquatilis BS-1 was able to generate a cold-adaptive enzyme. These two cold-adapted strains, and most notably the ${\beta}-galactosidase$ from each isolate, might prove useful in some biotechnological applications.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.10
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• pp.1397-1401
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• 2003

The aim of this study was to determine genotypes of four genetic markers and to investigate their association with milk production traits in Brown Swiss cattle imported to Slovakia. The bovine $\kappa$-casein, $\beta$-lactoglobulin, growth hormone and prolactin genotypes of 107 cows were identified by polymerase chain reaction. Effects all four genetic markers on milk, fat, protein and lactose yields and fat, protein and lactose percentage were estimated from a data set of 249 lactations. The frequency of desirable B allele of $\kappa$-casein gene to milk production was 0.46, alleles A of $\beta$-lactoglobulin gene was 0.55, allele and L of growth hormone gene was 0.45 and allele A and B of bovine prolactin gene were 0.61 and 0.39. The results of milk production obtained in our work showed that BB genotypes of $\kappa$-CN gene, AA genotypes of $\beta$-LG gene, LL genotypes of bGH gene were significantly associated with better milk production traits, mainly about the fat content. Association of a bovine prolactin genotypes with milk production were not found.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.318-324
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• 1998

A ${\beta}$-galactosidase with high transgalactosylic activity was purified from a Bacillus species, registered as KFCC10855. The enzyme preparation showed a single protein band corresponding to a molecular mass of 150 kDa on SDS-PAGE and gave a single peak with the estimated molecular mass of 250 kDa on Sephacryl S-300 gel filtration, suggesting that the enzyme is a homodimeric protein. The amino acid and sugar analyses revealed that the enzyme is a glycoprotein, containing 19.2 weight percent of sugar moieties, and is much more abundant in hydrophilic amino acid residues than in hydrophobic residues, the mole ratio being about 2:1. The pI and optimum pH were determined to be 5.0 and 6.0, respectively. Having a temperature optimum at $70^{\circ}C$ for the hydrolysis of lactose, the enzyme showed good thermal stability. The activity of the enzyme preparation was markedly increased by the presence of exogenous Mg (II) and was decreased by the addition of EDTA. Among the metal ions examined, the most severely inhibitory effect was seen with Ag (I) and Hg (II). Further, results of protein modification by various chemical reagents implied that 1 cysteine, 1 histidine, and 2 methionine residues occur in certain critical sites of the enzyme, most likely including the active site. Enzyme kinetic parameters, measured for both hydrolysis and transgalactosylation of lactose, indicated that the enzyme has an excellent catalytic efficiency for formation of the transgalactosylic products in reaction mixtures containing high concentrations of the substrate.

• Korean Journal of Food Science and Technology
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• v.21 no.6
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• pp.795-799
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• 1989

This study was carried out to hydrolyze lactose in commercial milk by ${\beta}-galactosidase$ from Kluyveromyces fragilis and to compare some physicochemical properties of yoghurts made from control and lactase-treated commercial milks. Quantitative analysis of sugars was performed by gas liquid chromatograph (GLC) on trimethylsilyl (TMS) derivatives. In commercial milk, 94.6% of lactose was hydrolyzed after 2 hours incubation at $40^{\circ}C$ with 6.0 units/ml of ${\beta}-galactosidase$. pH, titratable acidity and viable cell number of yoghurt made from lactase-hydrolyzed (LH) commercial milk were 4.1, 1.04% and $6.5{\times}10^8/ml$ of Str. thermophilus, $8.9{\times}10^8/ml$ of L. bulgaricus after 8 hours incubation at $40^{\circ}C$, respectively, The total contents of amino acid were 2.63% in control and 2.19%. in LH yoghurt. The total contents of free amino acid were 26.95 mg% in control and 17.55mg% in LH yoghurt. Analysis of free fatty acids resulted in that the contents of short chain fatty acids in LH yoghurt were a little higher than those in control. Both in control and LH yoghurt, the palmitic acid content was highest and that was followed by oleic and myristic acid.

• Korean Journal of Food Science and Technology
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• v.27 no.6
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• pp.878-884
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• 1995

In order to use whey lactose in alcohol fermentation, we investigated fermentation conditions of Saccharomyces cerevisiae and Kluyveromyces fragilis in lactose-hydrolyzed whey with ${\beta}-D-galactosidase$. and optimum conditions of the above two yeasts through oxygen regulation by Pasteur effect which is the characteristic of the yeasts were determined. In addition, optimum condition for application of fermented whey in Tak-ju process was also examined. With 0.7% ${\beta}-D-galactosidase$, 93% lactose was hydrolyzed at pH 6.5 in 30 minutes. Because S. cerevisiae is unable to ferment galactose, the production of ethanol by S. cerevisiae was lower than that of K. fragilis in lactose-hydrolyzed whey. But ethanol productivity by S. cerevisiae was higher than that by K. fragilis in glucose added whey. In fermentation with oxygen regulation and addition of 60 g/l glucose, the ethanol productivity of K. fragilis and S. cerevisiae were 18.9 g/l (11.8% increase) and 43.5 g/l (22.1% increase), respectively. It appeared that the ethanol productivity of S. cerevisiae was higher than thst of K. fragilis under the above conditions. In ethanol fermentation added rice starch, Aspegillus oryzae hydrolyzed 80% of starch in 60 hours, and the production of ethanol was 80.2 g/l

• Journal of Microbiology and Biotechnology
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• v.29 no.11
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• pp.1717-1728
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• 2019

The gene encoding β-galactosidase was cloned from Bifidobacterium longum RD47 with combinations of several bifidobacterial promoters, and expressed in B. bifidum BGN4. Among the recombinant bifidobacteria, BGN4+G1 showed the highest β-galactosidase level, for which the hydrolytic activity was continuously 2.5 to 4.2 times higher than that of BGN4 and 4.3 to 9.6 times higher than that of RD47. The β-galactosidase activity of BGN4+G1 was exceedingly superior to that of any of the other 35 lactic acid bacteria. When commercial whole milk and BGN4+G1 were reacted, BGN4+G1 removed nearly 50% of the lactose in the milk by the 63-h time point, and a final 61% at 93 h. These figures are about twice the lactose removal rate of conventional fermented milk. As for the reaction of commercial whole milk and crude enzyme extract from BGN4+G1, the β-galactosidase of BGN4+G1 eliminated 51% of the lactose in milk in 2 h. As shown below, we also compared the strengths and characteristics of the strong bifidobacterial promoters reported by previous studies.

• Archives of Pharmacal Research
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• v.21 no.4
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• pp.429-435
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• 1998

Lectins and its A- and B-chains from Korean mistletoe (Viscum album var. coloratum) were isolated by affinity chromatography on the Sepharose 4B modified by lactose-BSA conjugate synthesized by reductive amination of ligand (lactose) to .epsilon.-amino groups of lysine residues of spacer (BSA) after reduction by $NaCNBH_3$. The lactose-BSA conjugate was coupled to Sepharose 4B activated by cyanogen bromide. The molecular weight determined by SDS-PAGE were a 31 kD of A-chain and a 35kD of B-chain. Amino acid analysis and N-terminal sequencing were performed. The effects of pH, temperature and guanidine chloride on the conformation of the lectin were investigated by measuring its intrinsic fluorescence and compared with its hemagglutinating activities. Blue shift was detected on the acidic pH and there was a close relationship between activities and conformation of the lectin. Under denaturing conditions, the tryptophan emission profile of lectin showed typical denaturaiional red shift which also correspond to the conformations and activity of lectin.

• The Korean Journal of Food And Nutrition
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• v.12 no.1
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• pp.63-68
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• 1999

Production of galactooligosaccharides by an immobilized $\beta$-galactosidase from Aspergillus niger CAD 1 in sodium alginate was investigated. The ranges of temperature and pH for the maximum stability of im-mobilized $\beta$-galactosidase were 20~45$^{\circ}C$ and 4.0~5.5, respectively. The activation energy for the immob-illized $\beta$-galactosidase was 13,400 cal/mole At the concentration of the immobilized $\beta$-galactosidase 0.12 unit/g in sodium alginate the yield of galactooligosaccharides in cheese whey containing 20% lactose was 18% after incubation for 72 hr at 45$^{\circ}C$. The remaining activity for the immobilized $\beta$-galactosidase 10 times repeated use 87%.