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Production of protein-based polymers in Pichia pastoris

Werten, Marc W.T., Eggink, Gerrit, Cohen Stuart, Martien A., de Wolf, Frits A.
Biotechnology advances 2019 v.37 no.5 pp. 642-666
Escherichia coli, Pichia pastoris, amino acid sequences, biomedical research, bioprocessing, bioreactors, biosynthesis, collagen, elastin, genetic engineering, liquid state fermentation, molecular weight, nanomaterials, physical properties, polymers, process control, protein synthesis, proteolysis, recombinant DNA, silk, yeasts
Materials science and genetic engineering have joined forces over the last three decades in the development of so-called protein-based polymers. These are proteins, typically with repetitive amino acid sequences, that have such physical properties that they can be used as functional materials. Well-known natural examples are collagen, silk, and elastin, but also artificial sequences have been devised. These proteins can be produced in a suitable host via recombinant DNA technology, and it is this inherent control over monomer sequence and molecular size that renders this class of polymers of particular interest to the fields of nanomaterials and biomedical research. Traditionally, Escherichia coli has been the main workhorse for the production of these polymers, but the methylotrophic yeast Pichia pastoris is finding increased use in view of the often high yields and potential bioprocessing benefits. We here provide an overview of protein-based polymers produced in P. pastoris. We summarize their physicochemical properties, briefly note possible applications, and detail their biosynthesis. Some challenges that may be faced when using P. pastoris for polymer production are identified: (i) low yields and poor process control in shake flask cultures; i.e., the need for bioreactors, (ii) proteolytic degradation, and (iii) self-assembly in vivo. Strategies to overcome these challenges are discussed, which we anticipate will be of interest also to readers involved in protein expression in P. pastoris in general.