Part:BBa_K3784005

bg2 (Encoding β-galactosidase)

Patients with CKD are often accompanied by intestinal flora imbalance, and the number, composition, distribution and structure of intestinal flora have changed, which is manifested by the decrease of beneficial bacteria and the abnormal proliferation of pathogenic bacteria and conditional pathogens.

Therefore, we hope that our engineered bacteria can regulate the colony structure of the intestines, promote the proliferation of lactobacilli, bifidobacteria and other probiotics, and inhibit the proliferation of pathogenic bacteria such as Clostridium sporogenes. And galactooligosaccharide(GOS) is a very good choice. Studies have shown that galactooligosaccharide has a very good ability to promote the proliferation of intestinal probiotics such as bifidobacteria and lactobacilli, and it can inhibit the proliferation of intestinal pathogenic bacteria such as Clostridium sporogenes and reduce the production of intestinal endotoxins. At the same time, galactooligosaccharides can alleviate the intestinal inflammation caused by LPS stimulation and enhance the human intestinal barrier function.

We have heterologously expressed the β-galactosidase bga2 of strain Klebsiella oxytoca ZJUH1705 in the engineered bacteria. It has been confirmed that this enzyme has up to 45% of the ability to catalyze lactose to form galacto-oligosaccharides. This is a very high catalytic ability.

Characterization

To test whether our engineering bacteria can transcribe and encode normally the gene of β-galactosidase, we extracted the enzyme RNA for cDNA, and used it as template for PCR after agarose gel electrophoresis.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1708
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1708
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1708
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1708
  Illegal NgoMIV site found at 176
  Illegal NgoMIV site found at 1323
  
• 1000
  COMPATIBLE WITH RFC[1000]