Unit 6 Research Assignment

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Quiz 6 Research Assignment

My unknown bacteria was E. Coli which is a aerobe, mixed acid fermenting, gram negative bacteria. 3 biochemical tests I could have done during the process of identifying my unknown bacteria is the nitrate reductase test, ONPG Test and the MUG test.
The nitrate reduction test was one test we learned about in lab but did not use during the unknown identification process. This test helps identify a bacteria that produces the nitrate reductase enzyme which hydrolyze nitrate (NO3–) to nitrite (NO2–). The nitrate reduction test is an easy simple colometric test. If an organism can produce nitrate reductase in a solution rich in nitrate it will reduce the nitrate to nitrite and then the nitrites will form nitrous acid in solution. The first reagent called sulfanilic acid is then added to solution and it will react with the nitrous acid to produce diazotized sulfanilic acid. When the second reagent α-naphthylamine is added to solution it will react with diazotized sulfanilic acid …show more content…
Coli is the ONPG test. ONPG stands for O-Nitrophenyl-β-D-galactopyranoside which is an analog of lactose. The ONPG test allows the identification of lactose fermenting bacteria which contain the enzyme β-galactosidase. β-galactosidase can cleave the ONPG compound into galactose and o-nitrophenol. ONPG is a colorless compound and O-nitrophenol is yellow, therefore a visible color change indicates the successful hydrolysis of ONPG. This makes the ONPG test another simple colometric test. E. Coli is a lactose fermenting bacteria that contains β-galactosidase and therefore will be positive for the ONPG test. If an organism does not contain β-galactosidase the ONPG compound will not be hydrolyzed and no color change will be observed. This test requires the growth of unknown bacteria in a medium rich with lactose because β-galactosidase is only made by the bacteria in the presence of lactose. No other reagents are required. (Acharya,
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