Recombinant milk protein polymers
A polymer and milk protein technology, applied in mammalian protein, plasma globulin/lactoglobulin, animal protein processing, etc., can solve problems such as the difficulty of researching and predicting the properties of polymers in polymerization reactions
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment 1
[0184] Embodiment 1: the production of recombinant milk protein monomer in recombinant Kluyveromyces lactis host cell
[0185] Get bovine α-S1 casein (UniProt accession number P02662), bovine α-2 casein (UniProt accession number P02663), bovine β-casein (UniProt accession number P02666), bovine kappa-casein (UniProt Deposit No. P02668), bovine α-lactalbumin (UniProt Deposit No. B6V3I5) and bovine β-lactoglobulin (UniProt Deposit No. P02754) and were altered using the following changes: removal of 15 or 21- Residue signal peptide, add XhoI (CTC GAG) endonuclease recognition sequence and KEX endonuclease recognition sequence (AAA AGA) to the 5' end of DNA, and add SalI (GTC GAC) endonuclease recognition sequence to 3' end of DNA. Additional combined sequences were prepared by combining the sequences of the 4 caseins in the order shown above, and isolating each sequence using the following DNA fragments: GGC TCA GGA TCAGGG TCG AAA AGA GGC TCA GGA TCA GGG TCG, where non-un...
Embodiment 2
[0191] Embodiment 2: the production of recombinant milk protein monomer in recombinant Pichia host cell Recombinant Pichia host cells are produced by transforming the Komagataella phaffii strain BG12 (Biogrammatics, Carlsbad, CA) with a vector containing a European bovine milk protein (e.g., β-lactoglobulin) coding sequence that Codon-optimized for expression in yeast, and fused in-frame with an N-terminal secretion signal (pre or pre-pro signal peptide region of Saccharomyces cerevisiae alpha mating factor, native milk protein signal peptide, OST1 signal peptide). The milk protein coding sequence is flanked by promoters (constitutive promoters pGAP or pPGK, or methanol-inducible promoter pAOX1) and terminators (tAOX1 pA signal). The vector also contains a dominant resistance marker for selection of bacterial and yeast transformants and a bacterial origin of replication. This vector was transformed into Pichia host cells using a heat shock protocol to generate a recombinant ...
Embodiment 3
[0193] Example 3: Production of recombinant β-lactoglobulin monomer in recombinant Trichoderma reesei host cell
[0194] Recombinant T. reesei host cells were generated by transforming Hypocrea jecorina strain Qm6a with a vector containing the European bovine β-lactoglobulin coding sequence. Supernatant samples were used for ELISA assay and SDS-PAGE gel analysis to identify positive transformants.
[0195] Recombinant Trichoderma reesei host cells were grown in a stirred fermentation vessel at a temperature of 25°C-34°C, with aeration rates of 0.2 V / Vm and 1 V / Vm and minimal agitation, on a medium containing inorganic salts (as Minimal basis of phosphate, ammonium, magnesium, potassium, sodium, sulfate, chloride, calcium, iron, manganese, zinc, molybdenum, copper, cobalt and borate sources) with carbohydrate-based carbon sources as starting materials media to ensure proper mixing and dispersion of biomass and nutrients (including oxygen) (for delivery in compressed air). T...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More