Recombinant expression platform, constructs and methods for expression of difficult to express proteins (DTE-PS)

A versatile recombinant expression platform comprising:

• i. an engineered protease deficient yeast host cell with disrupted endogenous genes encoding protease PRB 1, protease PEP4 and auxotrophic markers uracil, lysine, adenine and leucine;
• and
• ii. an array of one or more episomal or integrated yeast based expression vectors operably linked with one or more promoters selected from Gal1 promoter, ADH2 promoter or Gal10 promoter, wherein the promoters can be used singly or in combination, said vector comprises a nucleic acid sequence having at least 90% sequence identity to at least one SEQ ID NO selected from:
• SEQ ID NOs:
• 7, 8, 9, 10, 11, and 12, which directs insertion of full length or truncated polynucleotide sequence into said yeast host cell for the expression of at least one target protein, and
• wherein said platform allows for enhanced expression of difficult to express target proteins of diverse origin and families compared to a control yeast host cell expression.