Advances in Cell Engineering of the Komagataella phaffii Platform for Recombinant Protein Production

Abstract

Komagataella phaffii (formerly known as Pichia pastoris) has become an increasingly important microorganism for recombinant protein production. This yeast species has gained high interest in an industrial setting for the production of a wide range of proteins, including enzymes and biopharmaceuticals. During the last decades, relevant bioprocess progress has been achieved in order to increase recombinant protein productivity and to reduce production costs. More recently, the improvement of cell features and performance has also been considered for this aim, and promising strategies with a direct and substantial impact on protein productivity have been reported. In this review, cell engineering approaches including metabolic engineering and energy supply, transcription factor modulation, and manipulation of routes involved in folding and secretion of recombinant protein are discussed. A lack of studies performed at the higher-scale bioreactor involving optimisation of cultivation parameters is also evidenced, which highlights new research aims to be considered.

Role of Dissimilative Pathway of Komagataella phaffii (Pichia pastoris): Formaldehyde Toxicity and Energy Metabolism

“…Komagataella phaffii (formerly known as Pichia pastoris ) is a methylotrophic yeast extensively used for recombinant protein production (See [ 1 , 2 ] for recent reviews). In this microorganism, methanol induces the peroxisome biosynthesis and expression of genes encoding enzymes required for its metabolism [ 3 ].…”

Codon-tRNA Coadaptation Bias for Identifying Strong Native Promoters in Komagataella phaffii

“…Challenges related to methanol use (e.g., fire hazard, inducer toxicity, a high catabolic oxygen requirement leading to high heat production) have motivated the discovery of alternative promoters for methanol-free heterologous expression in K. phaff ii. 3 The constitutive promoter of the Glyceraldehyde-3-phosphate dehydrogenase (GAP) gene (P GAP ) represents another K. phaff ii benchmark promoter. 4 Apart from overcoming challenges associated with methanol use and transitioning to a more scaleup friendly substrate (i.e., glucose), P GAP bypasses potential drawbacks associated with inducible systems (decouple growth and production phases), which might be cumbersome when scaling up in continuous cultivations.…”

“…Since promoters are critical for controlling the timing and scale of production, an appropriate promoter must be chosen for optimal bioprocess performance. Challenges related to methanol use (e.g., fire hazard, inducer toxicity, a high catabolic oxygen requirement leading to high heat production) have motivated the discovery of alternative promoters for methanol-free heterologous expression in K. phaffii …”

Industrial Production of Proteins with Pichia pastoris—Komagataella phaffii

“…Komagataella phaffii ( K. phaffii ; previously described as Pichia pastoris ) is a yeast strain relevant to the industrial production of proteins. This species is widely applied as a heterologous protein production host, and its utilization has been widely reported in the literature [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 ]. The main advantages of this organism are the possibility to run high-density fermentation according to established protocols, fast-paced and automation-friendly genetic engineering [ 10 ], eukaryotic post-translational modifications [ 11 , 12 ], high secretory efficiency and biomass yields [ 13 , 14 ], stable genetic constructs [ 15 ], and an increasing collection of publicly available tools [ 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ].…”