Elsevier

Biotechnology Advances

Volume 37, Issue 5, September–October 2019, Pages 642-666
Biotechnology Advances

Research review paper
Production of protein-based polymers in Pichia pastoris

https://doi.org/10.1016/j.biotechadv.2019.03.012Get rights and content
Under a Creative Commons license
open access

Abstract

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.

Keywords

Pichia pastoris
Protein-based polymers
Protein expression
Proteolysis
Silk
Collagen
Gelatin
Elastin
Block copolymers
Self-assembly

Abbreviations

AOX1
alcohol oxidase 1
BiP
immunoglobulin-binding protein
DPAPase A
dipeptidyl aminopeptidase A
EBP
elastin-binding protein
ECM
extracellular matrix
ELP
elastin-like polypeptide
ERAD
ER-associated degradation
FDA
food and drug administration
GAP
glyceraldehyde-3-phosphate dehydrogenase
GRAS
generally recognized as safe
his4
histidinol dehydrogenase
ITC
inverse transition cycling
LCST
lower critical solution temperature
MaSp1
major ampullate spidroin 1
MaSp2
major ampullate spidroin 2
OD600
optical density at 600 nm
P4H
peptidyl-prolyl-4-hydroxylase
PDI
protein disulfide isomerase
Pho1
acid phosphatase
ppαF
Saccharomyces cerevisiae α-factor mating pheromone prepro peptide
UPR
unfolded protein response

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