Part:BBa_K2711000

Glucanase

β-Glucan is a water-soluble polysaccharide that consists of glucose units. In yeast cell walls, glucose monomers are linked in a branched structure via β-(1→3) and β-(1→6) glycosidic bonds [1]. β-1,3-Glucanases (EC 3.2.1.39 and EC 3.2.1.6) catalyze the hydrolysis of glucans containing β-1,3-linked glucose monomers, one of the most abundant polysaccharides present in the yeast cell wall [2]. Bacterial endo-1,3-β-glucanases catalyze the hydrolysis of (13)-beta-D-glucosidic linkages in (1 → 3)-beta-D-glucans. These enzymes have a β-sandwich architecture and belong to the glycoside hydrolase family 16 (GH16) [3].

The UiOslo_Norway 2018 team used the sequence of endo-1,3-β-glucanase from Cellulosimicrobium cellulans (= Arthrobacter Luteus) to enable targeted lysis of yeast cells. This glucanase is also used in the commercially available product Zymolyase, among other components, which is a lytic enzyme for the digestion of yeast cell wall glucan. In our case we wanted to specifically lyse Candida albicans cells in a vaginal sample which will also contain for example bacteria and skin cells. The glucanase was synthesized by IDT after the sequence was codon optimized.

SOURCES:

1. Deepak Mudgil, in Dietary Fiber for the Prevention of Cardiovascular Disease, 2017

2. Salazar et al., 2001. Overproduction, Purification, and Characterization of β-1,3- Glucanase Type II in Escherichia coli. Protein Expression and Purification 23, 219–225.

3. Oda et al., 2017. Structural and thermodynamic characterization of endo-1,3-β-glucanase: Insights into the substrate recognition mechanism. BBA - Proteins and Proteomics 1866 (2018) 415–425.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 958
  
• 1000
  COMPATIBLE WITH RFC[1000]