Difference between revisions of "Part:BBa K4309000"
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| + | Fig.1. SDS-PAGE analysis of AE5 mutant strain CotA . SDS-PAGE was used to analyze the expression of CotA. Recombinant vectors pHJ5 transformed into BL21 (DE3) competent cells and induced by 0.1 mM IPTG in LB medium for 20 h at 16 ℃ , respectively. All the samples were analyzed by SDS-PAGE, and the protein was stained with Coomassie Blue in the gel. Lane M, protein marker. Lane 1-2, whole bacterial lysate of the E.coli BL21 (DE3) contained recombinant pET28a-cotA which was induced. . Lane 5-6, whole bacterial lysate of the E.coli BL21 (DE3) containing empty pET28a. S: Supernant; P: Pellet. | ||
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| + | We changed the temperature and the concentration of the inducer. The figure above clearly shows that the target protein induced in LB medium is present in the supernatant (Fig. 1). | ||
Revision as of 15:02, 11 October 2022
Laccase（Bacillus）
Lignin peroxidase (LiP), phenoloxidases (laccases, tyrosinases), manganese dependent peroxidase (MnP) are the three enzymes commonly employed ligninolytic enzymes which are mainly involved in degrading lignin and analogue PAHs. Various researches have revealed that the mechanism of oxidation of PAHs by fungi ligninolytic enzymes is similar to the degradation of nonphenolic lignin.In vitro,laccases has activity towards 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxy-phenol, and guaiacol.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1207
Illegal AgeI site found at 1348 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1275
Illegal BsaI.rc site found at 1509
Illegal SapI.rc site found at 286
Fig.1. SDS-PAGE analysis of AE5 mutant strain CotA . SDS-PAGE was used to analyze the expression of CotA. Recombinant vectors pHJ5 transformed into BL21 (DE3) competent cells and induced by 0.1 mM IPTG in LB medium for 20 h at 16 ℃ , respectively. All the samples were analyzed by SDS-PAGE, and the protein was stained with Coomassie Blue in the gel. Lane M, protein marker. Lane 1-2, whole bacterial lysate of the E.coli BL21 (DE3) contained recombinant pET28a-cotA which was induced. . Lane 5-6, whole bacterial lysate of the E.coli BL21 (DE3) containing empty pET28a. S: Supernant; P: Pellet.
We changed the temperature and the concentration of the inducer. The figure above clearly shows that the target protein induced in LB medium is present in the supernatant (Fig. 1).