Synthesis and secretion of hepatitis B middle surface antigen by the methylotrophic yeast Hansenula polymorpha.
The methylotrophic yeast, Hansenula polymorpha, has been developed as a host system for the synthesis of heterologous proteins. The middle surface antigen of hepatitis B virus (preS2-HBsAg) has been synthesized under the control of a methanol-regulated promoter derived from the methanol oxidase-encoding gene. The synthesized preS2-HBsAg protein was found to be secreted outside the cell membrane into the periplasm and further excreted into the culture medium following permeabilization of the cell wall with beta-1,3-glucanase (beta Glu). Cell cultures treated with beta Glu were able to continuously synthesize and secrete 22-nm particles of preS2-HBsAg into the medium for several days. The overall yield of antigen from treated cultures was found to be over threefold greater than that of untreated controls. The observation that complex supramolecular structures, such as the 22-nm particles of preS2-HBsAg, can be secreted by H. polymorpha and released into the medium, suggests the potential for these yeasts to be an alternative secretory host.
Shen
S H
SH
Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec.
Bastien
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L
Nguyen
T
T
Fung
M
M
Slilaty
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SN
D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D043326:Glucan 1,3-beta-Glucosidase; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000378:metabolism; D015151:Immunoblotting; D010843:Pichia / Q000235:genetics*; D010957:Plasmids; D011498:Protein Precursors / Q000096:biosynthesis / Q000378:metabolism / Q000648:ultrastructure; D004718:Saccharomycetales / Q000235:genetics*; D014170:Transformation, Genetic; D001617:beta-Glucosidase
D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase