Coding

Part:BBa_K3196003

Designed by: Hao Qiu   Group: iGEM19_HUST-China   (2019-10-16)


pelA

Synthetic Aspergillus pectin lyase to decompose pectin into galacturonic acid.

Characterization

Pectinase is a generic term for a class of enzymes that break down pectin and it is an important industrial enzyme.

Usage and Biology

Pectinase is widely used in the food industry, such as juice and fruit wine processing, hemp degumming, wood preservation, biopulping, environmental protection, soil softening treatment and feed industries. The degradation of pectin substances in recent years has attracted widespread attention from scholars at home and abroad, now there is a more in-depth study on the mechanism of action of various components of pectinase, regulation of gene expression, and selection of constitutive strains.[1] Pectinase is a general term for a class of enzymes that break down pectin, among them, the lyase can cleave the glycosidic bond in the highly methylated interior, and does not require the pre-action of pectinase, and is an important enzyme in the degradation process of pectin. The pectin lyase gene cloned from EIM-6 Aspergillus niger. Preliminary purification results indicated that the main components of Aspergillus niger EIM-6 pectinase were lyase and galacturonase. [2]


DNA Gel Electroghoretic

Electro-transformation to Pichia pastoris was all succeeded.

Figure1 Yeast Genome extraction and pcr

Enzyme Activity

The enzyme activity of the pelA group is obtained by titration and daily measurement of enzyme activity.

Figure2 Determination of pelA enzyme activity


Pectin is a hydrophilic polysaccharide substance present in the interstitial of plant cells. It is stable in acidic medium and insoluble in ethanol. It is a natural polymer compound. We understand that pectinase measures enzyme activity has a nationally standardized experimental procedure, so we have adopted the standard titration scheme given by the state. [3]


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 745
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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