LOG Enzyme Expression and Purification

The coding region from CvarLOG1 was amplified from Chlorella variabilis cDNA. The primers used for the PCR were LOG pET28 F and LOG pET28 R (see Supplementary Table 5). The PCR products were ligated into the pET28a (Novagen) to express His-tagged recombinant proteins. The BL21 (DE3) strain was used to express the recombinant protein that was induced in LB broth with 1 mM isopropyl-D-thiogalactopyranoside for 5 h at 37°C. After induction, Escherichia coli cells were pelleted and stored at −20°C until further use. The protein was extracted using the Capturem His-Tagged Purification Miniprep Kit (Takara Bio, Japan) to purify recombinant proteins according to the manufacturer’s protocol.

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