Part:BBa_K2377003

pectinase

Different types of pectic enzymes vary in how they degrade pectin, a long polysaccharide of sugars that forms a gel. Pectin forms the center of the plant cell wall. Other molecules, like cellulose, are embedded in it. When pectin is degraded by pectinase, the cell walls become weaker. We want to use pectinase and celluse in our downstream circuits, to let them woking together in causing minor wound on the leave of plants. And test wether dsRNA could enter leave through these wounds inducing plant RNAi. But owing to time deficiency, this integrated effect havn't be demonstrated . We will complete it our future work.

Lethbridge_HS contribution 2020: Literature Report

Recent literature has shown that pectinases can be produced in large scale batches using organisms such as Aspergillus niger using an optimal growth media [1]. However most of these pectinases produced for commercial purposes are active only at about 50˚C [2]. Some pectinases found in plant leaf tissue can provide more stable products but there are still limitations to thermostable options [3]. These industrial pectinase enzymes are commonly purified using ammonium salt precipitation, gel filtration chromatography and ion exchange chromatography [4]. The pectinase represented from this part is a pectinase enzyme from Alkalihalobacillus halodurans or also known as Bacillus halodurans. Since the making of this part, there has been only one publication on this particular pectinase, published in 2013. Mei et al., [5] were able to clone, purify by affinity chromatography, and test the biochemical properties of this enzyme. The M29 strain of A. halodurans is an aerobic alkaliphilic bacterium that can handle temperatures up to 65˚C. The identified 39 kDa pectinase was cloned into E. coli for purification and characterization. This protein was determined to have optimal activity at pH 10 and a temperature of 80˚C. It was also determined that calcium was a necessary cofactor for enzymatic activity

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 87
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 576