Part:BBa_K4382006

Endo Pectin Lyase - PelB-B2

Usage

The PelB-B2 was obtained through a Bacillus sp.(OR-B2) identified during metagenomic analysis of orange peel. PelB-B2 is a pectin lyase - i.e. it helps breakdown pectin into a 4,5-unsaturated galacturonides. Pectin breakdown is quite useful in various industrial applications such as retting of fibers, clarifying juices, paper making etc.

Biology

The optimum temperature and pH at which PelB-B2 works is 500C and at a pH of 8, although it is highly thermostable and works at a very large range of pH (i.e. between 6 to 11). An important property of this pectinase is that its catalytic activity is significantly enhanced by Ca2+ and reduced by chelating agents such as EDTA, SDS and metal ions such as Hg2+, Ag2+ and Co2+. While it prefers apple and citrus peels as substrates, it also breaks down commercially available polygalacturonases with reasonably good efficiency.

The activity of this enzyme was measured using a DNS assay. Briefly, pectin was incubated with the PelB-B2. After 30 minutes of incubation, DNS (3,5-dinitrosalicylic acid) was added and boiled for 10 minutes in order to generate color. Following this, OD540 was measured and the value was plotted on a standard curve prepared using commercially available D-(+)-Galactouronic acid.

References

1. Guan, Y., Wang, D., Lv, C., Zhang, Y., Gelbic, I., & Ye, X. (2020, January 13). Archives of microbiology: screening of pectinase-producing bacteria from citrus peel and characterization of a recombinant pectate lyase with applied potential. Archives of Microbiology, 202(5), 1005–1013. https://doi.org/10.1007/s00203-020-01807-0 2. De Prez, J., Van Vuure, A. W., Ivens, J., Aerts, G., & Van de Voorde, I. (2018, December). Enzymatic treatment of flax for use in composites. Biotechnology Reports, 20, e00294. https://doi.org/10.1016/j.btre.2018.e00294

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]

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