Part:BBa_K3196002

SLAC

Usage and Biology

Laccase is a multicopper oxidase, capable of catalyzing one-electron oxidation of a wide range of substrates to generate radicals while concomitantly reducing molecular oxygen to water^[1]. Laccases are widely distributed in nature. They have been isolated from fungi, plants, insects and bacteria^[2] of which fungal laccases are well characterized ^[3]. Bacterial laccases, such as the small laccase from S. coelicolor (SLAC), can be useful alternatives to fungal laccases. SLAC exhibits some desirable characteristics such as activity at high temperatures and alkaline pH ^[4], making it a potential candidate in biotechnological applications. Extracellular expression could be a desirable alternative for expression of foreign proteins since it simplifies purification steps ^[5]. P. pastoris is usually the preferred host for the production of industrial enzymes.  

DNA Gel Electrophoretic

Figure:This is the result of the SacⅠ single endonuclease digestion plasmid of the TOP10 strain, we will extract the gel and obtain the aimed DNA.

The image above shows the successful transformation of the target gene into the Pichia pastoris genome, including SLAC FLO10, SLAC PHO5 APRO, and SLAC.

SDS-PAGE

Figure:the SDS-page result shows that Pichia pastoris have successfully secrete SLAC protein into superfluous liquid.

Enzyme Activity

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 345