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Introduction
Tomlin, Lowis and Read (1991) indicate that many people exhibit clinical signs that are associated with constipation, anal inflammation, hemorrhoids, diarrhea and irritable bowel syndrome. According to Cristtofaro, Mottu and Wuhrmann (1974), other patients suffer from anemia and nutrient mal-absorption which predisposes secondary infections and illnesses. These clinical signs are brought about by food intolerance especially dairy food intolerance subject to lactose intolerance hose prevalence is three for every four people, sensitivity to yeast which is secondary to Candida infections with a prevalence of one in every three people, sensitivity to gluten that is secondary to celiac and wheat intolerance and fructose or sugar intolerance that has a prevalence of one in every three people (Tomlin, Lowis and Read 1991: Ganiats, T., Norcross, W. Halverson, et al, 1994). The flatus is brought about by flatulence events that are dependent on breakdown of oligosaccharides by yeast or human symbiotic bacteria that reside in the large intestine (Bullerwell, Raunig and Hagar, 1994).
Statement of the problem
This experiment is designed to investigate volume of flatus released or produced within a period of two hours relative. The experiment investigates effect of PH on bio-engineered enzyme alpha galactosidase enzyme.
Methodology of the experiment
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Chemical ingredients required
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Bean solution
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Yeast solution
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PH buffers
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Equipments required
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PH meter
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Measuring cylinder
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Mortar
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Pestle
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Bunsen burner and its associated apparatus (gas cylinder, tripod stand, wire gauze)
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Water bath and its associated apparatus (thermostat, water heater, plugs and source of electricity)
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Test tube holders
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Match box
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Universal indicator
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Universal PH meter
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Cheese cloth
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Preparation of ingredients
Preparation of the bean solution
Ten grams of Beano beans were soaked for 24 hours in distilled water. After 24 hours, water was poured off and the beans were pulverized and ground using mortar and pestle. The slurry was added into 100 ml of water and heated to boiling for three minutes. After three minutes, the heat source was removed and the slurry allowed to cool gradually. Once it had attained the slurry had attained a temperature below the room temperature that was maintained for more than six hours, the slurry was filtered through a cheese cloth and filtrate stored in a labeled test tube for fermentation experiment.
Preparation of yeast solution
Ten grams of bakers yeast (Saccgarintces cerevisiae Fleishmanns active dry yeast, YSC 2 from Sigma) was added into 100 ml of distilled water and placed in 37degrees celcius thermostated conditions for 1.5 hours.
Preparation of buffer solutions
PH buffer solutions were made from standard stocks and their PH ranged from PH 2 to PH 10.
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Experimental procedure
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Step 1: 9 smith fermentation dry test tubes were labeled and 10ml of Yeast with alpha galactosidase enzyme was measured using a 10ml measuring cylinder.
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Step 2: 9 smith fermentation dry test tubes were labeled and 10ml of Yeast without alpha galactosidase enzyme was measured using a 10ml measuring cylinder and put into a test tube.
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Step 3: each of the nine smith fermentation test tubes in step 2 and step 3 were adjusted to a particular PH ranging from PH 2 to PH 10 using 9 ml of buffer solution
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Step 4: 5 ml of bean solution was added and 4 ml of yeast solution added to respective smith fermentation test tubes
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note: for each two test tubes at a given PH like PH 2, both tubes will have bean solution and buffer, but only one tube will have five drops of Beano added.
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Step 5: Add Beano to selected fermentation tubes and mix thoroughly. After mixing, place the Smith tubes in the 37 Centigrade water bath.
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Step 6: take readings of gas evolved or released in an interval of ten minutes for two hours.
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Step 7: record the maximum volume of gas evolved at the end of the two hour period.
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Results
Table 1: combined data of results obtained after two hours showing relationship between activities of the enzyme at different PH scales
Discussion of the results
Analysis of the mixture of slurry, yeast and Beano
From the results, it is evident that activity of the enzyme present in slurry, yeast and Beano mixture increases with increasing PH from PH 2 to PH 4 where the enzyme activity is optimum. As PH increases from PH 4, activity of the Enzyme decreases because increase in PH denatures the enzyme by interfering with the tertiary structure of the enzyme. Between PH 7 and PH 8, there is a sharp drop in Enzyme activity and a similar sharp rise in enzyme activity between PH 8 and PH 9.
Analysis of the mixture of slurry, yeast and Bean
The trend shows that there is decrease in activity of the enzyme from PH 2 to PH 8. As PH increases from PH 8 to PH 9, there is a sharp increase in activity of the enzyme followed by a Sharp decrease in enzyme activity between PH 9 and PH 10.
Differences between enzyme activity between mixture of slurry, yeast and Beano and mixture of slurry, yeast and bean
The enzyme activity in the mixture of slurry, yeast and Beano is higher than the enzymatic activity of the mixture of slurry, yeast and bean for all ranges of PH. As PH increases, the enzymatic activity of the mixture of slurry, yeast and beano increases while that of slurry, yeast and bean decreases indicating an inverse activity relationship.
Similarities between activities of enzymes in both samples
It is evident from the results that enzymatic activity has an optimum point at PH 8 for both tests.
Conclusion
Beano has a potential of increasing breakdown oligosaccharides into its building units. It therefore can be used to subsidize yeasts hydrolysis of oligosaccharides. The process leads into production of a lot of flatus and this could lead into noises that arise as a result of flatulence subject to vibrations of the anal sphincter. Therefore Beano promotes flatulence while bean does not support flatulence. It can be argues that bean has smoothing effect on the flatulence. From the results, bean contains digestive enzymes that diminish amount of flatulence caused by some components of foods that fail to be digested by the body hence promote action of microbes in the small and large intestines.
Future research work
Further studies should be geared towards investigating if Beano is effective in reducing the volume and frequency of flatus. Other future studies should be aimed at determining if agents that have lower surface tension could reduce effects of flatulence by improving gaseous dissolution into liquid and faecal matter.
Bibliography
Bullerwell, L., Raunig, V., & Hagar, W. (1994) Investigating a Bio-engineered enzyme-milking more from your enzyme laboratory The Science Teacher, 61, 27-29.
Ganiats, T., Norcross, W. Halverson, et al (1994) Does Beano prevent gas? A double blind crossover study of oral alpha galactosidase to treat dietary oligosaccharide intolerance J family practice, 39 441-445.
Tomlin, J., Lowis, C., & Read, N.W. (1991) investigation of normal flatus production in health volunteers, Gut, 32, 665-669.
Cristtofaro, E., Mutto, F. & Wuhrmann (1974). Sugars in nutrition . New York, NY, Academic press, 313-336.
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