Difference between revisions of "Part:BBa K2302006"
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Revision as of 12:44, 27 October 2017
CALB,an enzyme that can hydrolyze fat to fatty acids.
this sequence encodes a functional protein that can hydrolyze fat to fatty acids. The protein is one of the most widely used lipases due to its excellent properties, such as high stereoselectivity and stability.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 949
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 670
Illegal NgoMIV site found at 780
Illegal AgeI site found at 472
Illegal AgeI site found at 730 - 1000COMPATIBLE WITH RFC[1000]
RESULT
To further determine whether the recombinant CALB is functional, we made soft agar containing glyceryl tributyrate on culture plates containing growing bacteria (Fig.1). The result showed that Follower D could secrete degrade glyceryl tributyrate as indicated by transparent ring shaped as “NEFU”, suggesting its production of functional lipase.
Fig.1 The forming transparent area shaped as “NEFU”
Next, we used the culture supernatant of the bacteria to determine whether the lipase can be successfully secreted into culture medium. As shown in Fig.2, the culture supernatant could also cause degradation of glyceryl tributyrate and form transparent arear shaped as “IGEM”. The medium was stained by Sudan III. This result indicate that FD could indeed secret functional lipase into the culture medium, as we expected.
Fig.2 The forming transparent area shaped as “IGEM”.