Endogenous prolyl 4-hydroxylation in Hansenula polymorpha and its use for the production of hydroxylated recombinant gelatin.
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.
de Bruin
Eric C
EC
Agrotechnological Research Institute, Wageningen, The Netherlands.
Werten
Marc W T
MW
Laane
Colja
C
de Wolf
Frits A
FA
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*
D003470:Culture Media; D005656:Fungal Proteins; D011994:Recombinant Proteins; D005780:Gelatin; D003094:Collagen; D011392:Proline; D011393:Procollagen-Proline Dioxygenase