title: Endogenous prolyl 4-hydroxylation in Hansenula polymorpha and its use for the production of hydroxylated recombinant gelatin.
abstract: Several yeast systems have recently been developed for the recombinant production of gelatin and collagen. Amino acid sequence-specific prolyl 4-hydroxylation is essential for the gel-forming capacity of gelatin and for the proper folding of (pro)collagen. This post-translational modification is generally considered to be absent in microbial eukaryotic systems and therefore co-expression of heterologous (human or animal) prolyl 4-hydroxylase would be required. However, we found that the well-known protein expression host Hansenula polymorpha unexpectedly does have the endogenous capacity for prolyl 4-hydroxylation. Without co-expression of a heterologous prolyl 4-hydroxylase, both an endogenous collagen-like protein and a heterologously expressed collagen fragment were found to be sequence-specifically hydroxylated.
authors.affiliation: Agrotechnological Research Institute, Wageningen, The Netherlands.
authors.forename: Eric C
authors.initials: EC
authors.lastname: de Bruin
authors.forename: Marc W T
authors.initials: MW
authors.lastname: Werten
authors.forename: Colja
authors.initials: C
authors.lastname: Laane
authors.forename: Frits A
authors.initials: FA
authors.lastname: de Wolf
mesh_terms: D000595:Amino Acid Sequence; D019149:Bioreactors; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005780:Gelatin / Q000235:genetics / Q000378:metabolism*; D006900:Hydroxylation; D010843:Pichia / Q000201:enzymology* / Q000378:metabolism; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D011994:Recombinant Proteins; D005780:Gelatin; D003094:Collagen; D011392:Proline; D011393:Procollagen-Proline Dioxygenase