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Prebiotics Production Using Recombinant B-Galactosidase And Their Effect On Growth Of Rat Colon

By: Mubbashir Hassan Khan | Sr. Sanaullah Iqbal.
Contributor(s): Dr. Muhammad | Dr. Naureen Naeem.
Material type: materialTypeLabelBookPublisher: 2013Subject(s): Department of Food Sciences & HumanDDC classification: 1833,T Dissertation note: In this research work the prebiotic was prepared by using the enzyme from Kluyveromyces lactis and different doses were fed to the rats to check the growth of lactobacillus, bifidobacterium and E. coli. Firstly the enzyme activity was checked at different concentrations at different time intervals by using 20g/l lactose solution. The glucose, galactose and lactose and their mixture were used as a standard in the same concentration. After many thin layer chromatography analysis the 10µl enzyme for 45 min was enough to convert 1ml of the lactose solution. Then, by using the same concentrations of the enzyme prebiotics galactooligosaccharides were synthesized by transgalactosylation and were fed orally to group 1 for 10 days, group 2 for 20 days and group 3 for 30 days at concentrations of 1g/day, 2g/day and 3g/day respectively. After 10, 20 and 30 days the fecal samples from all the groups were collected aseptically to check the growth of lactobacillus, bifidobacterium and E. coli. Colony counting was done to count the colonies. In the study different doses of prebiotics GOS were given @ 1g/day, 2 g/day and 3 g/day. The plates were incubated at 37 ºC and numbers of colonies were counted on digital colony counter. The effect of different concentrations of dose of galactooligosaccharides and the effect of time duration was calculated. The P-value used was 0.05. The P-value obtained by the two way ANOVA for the effect of dose of different concentrations is 0.354521 which is more than 0.05. This value indicates that the effect of dose on lactobacillus growth is insignificant as for as the concentration is concerned. As the concentration is increased there is no effect on growth. While the other factor was the time duration i. e 10, 20 and 30 days so its effect is calculated. P-value obtained for the effect of time is 0.029958. It shows that the result is significant. As the time interval is increased the CFU/ml of lactobacillus also increased. The consequence of different concentration of dosage of prebiotic galactooligosaccharides (GOS) and the effect of time period is determined. 0.05 P-value used. The P-value deducted by using two-way ANOVA for the effect of dose of different concentrations is 0.032328 which is less than 0.05. This value indicates that the effect of dose on bifidobacteria growth is significant. As the concentration is increased there is increasing effect on growth. While the other factor was the time duration i. e 10, 20 and 30 days so its effect is determined. P-value obtained for the effect of time is 0.014286. It shows that the result is significant. As the time interval is increased the CFU/ml of bifidobacteria also increased. The consequence of different concentration of dosage of prebiotic galactooligosaccharides (GOS) and the effect of time period is determined for E. coli. The P-value obtained 0.03984 which is less than 0.05. This value indicates that the effect of dose on E. coli growth is significant. As the concentration is increased there is increasing effect on growth. P-value obtained for the effect of time is 0.004848. It shows that the result is significant. As the time interval is increased the CFU/ml of bifidobacteria also increased.
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In this research work the prebiotic was prepared by using the enzyme from Kluyveromyces lactis and different doses were fed to the rats to check the growth of lactobacillus, bifidobacterium and E. coli. Firstly the enzyme activity was checked at different concentrations at different time intervals by using 20g/l lactose solution. The glucose, galactose and lactose and their mixture were used as a standard in the same concentration. After many thin layer chromatography analysis the 10µl enzyme for 45 min was enough to convert 1ml of the lactose solution.
Then, by using the same concentrations of the enzyme prebiotics galactooligosaccharides were synthesized by transgalactosylation and were fed orally to group 1 for 10 days, group 2 for 20 days and group 3 for 30 days at concentrations of 1g/day, 2g/day and 3g/day respectively. After 10, 20 and 30 days the fecal samples from all the groups were collected aseptically to check the growth of lactobacillus, bifidobacterium and E. coli. Colony counting was done to count the colonies.
In the study different doses of prebiotics GOS were given @ 1g/day, 2 g/day and 3 g/day. The plates were incubated at 37 ºC and numbers of colonies were counted on digital colony counter. The effect of different concentrations of dose of galactooligosaccharides and the effect of time duration was calculated. The P-value used was 0.05. The P-value obtained by the two way ANOVA for the effect of dose of different concentrations is 0.354521 which is more than 0.05. This value indicates that the effect of dose on lactobacillus growth is insignificant as for as the concentration is concerned. As the concentration is increased there is no effect on growth. While the other factor was the time duration i. e 10, 20 and 30 days so its effect is calculated. P-value obtained for the effect of time is 0.029958. It shows that the result is significant. As the time interval is increased the CFU/ml of lactobacillus also increased.
The consequence of different concentration of dosage of prebiotic galactooligosaccharides (GOS) and the effect of time period is determined. 0.05 P-value used. The P-value deducted by using two-way ANOVA for the effect of dose of different concentrations is 0.032328 which is less than 0.05. This value indicates that the effect of dose on bifidobacteria growth is significant. As the concentration is increased there is increasing effect on growth. While the other factor was the time duration i. e 10, 20 and 30 days so its effect is determined. P-value obtained for the effect of time is 0.014286. It shows that the result is significant. As the time interval is increased the CFU/ml of bifidobacteria also increased. The consequence of different concentration of dosage of prebiotic galactooligosaccharides (GOS) and the effect of time period is determined for E. coli. The P-value obtained 0.03984 which is less than 0.05. This value indicates that the effect of dose on E. coli growth is significant. As the concentration is increased there is increasing effect on growth. P-value obtained for the effect of time is 0.004848. It shows that the result is significant. As the time interval is increased the CFU/ml of bifidobacteria also increased.

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