Parts
BBa_K4224000 Basic part - Aspergillus oryzae beta-galactosidase (lacA)
Aspergillus oryzae beta-Galactosidase (Ao-ß-gal) is an extracellular enzyme belonging to the GH35 family of beta-galactosidase (lactase). The amino acid sequence of Ao-ß-gal includes 1005 residues with an average molecular mass of 120.15kDa[9]. Ao-ß-gal has an acidic pH optimum of about 5.0 and high culture yield, less contaminated proteins, and easier downstream processing. In this project, we aimed at expressing Ao-ß-gal in Pichia pastoris X33.
Fig. 1 | The three-dimensional structure of the Ao-ß-gal presented as a ribbon model. The catalytic domain is colored in red.
BBa_K4224001 Composite part - AOX1 promoter+alpha factor signal+lacA+AOX1 terminator
This composite part is a part of vector pPICZ-alpha A plasmid for expressing Aspergillus oryzae beta-galactosidase in Pichia pastoris. This composite part contains a methanol-inducible promoter AOX1, an alpha factor signal containing rbs, the lacA gene from Aspergillus oryzae, and an AOX1 terminator.
Fig. 2 | The composite part of AOX1 promoter-alpha factor signal-lacA-AOX1 terminator
References
[1] Maksimainen MM, Lampio A, Mertanen M, Turunen O, Rouvinen J. The crystal structure of acidic β-galactosidase from Aspergillus oryzae. Int J Biol Macromol. 2013 Sep;60:109-15. doi: 10.1016/j.ijbiomac.2013.05.003. Epub 2013 May 17. PMID: 23688418.
[2] Shi, Xin, et al. "Engineering the optimum pH of β-galactosidase from Aspergillus oryzae for efficient hydrolysis of lactose." Journal of Dairy Science 105.6 (2022): 4772-4782.