Process technology for production and recovery of heterologous proteins with Pichia pastoris.
Developments in process techniques for production and recovery of heterologous proteins with Pichia pastoris are presented. Limitations for the standard techniques are described, and alternative techniques that solve the limitations problems are reviewed together with the methods that resulted in higher productivity of the P. pastoris processes. The main limitations are proteolysis of the secreted products and cell death in the high cell density bioreactor cultures. As a consequence, both low productivity and lower quality of the feedstock for downstream processing are achieved in processes hampered with these problems. Methods for exploring proteolysis and cell death are also presented. Solving the problems makes the conditions for downstream processing superior for the P. pastoris expression systems compared to other systems, which either need complex media or rely on intracellular production. These improved conditions allow for interfacing of cultivation with downstream processing in an integrated fashion.
Jahic
Mehmedalija
M
Department of Bioprocess Technology, School of Biotechnology, Royal Institute of Technology (KTH), S 10691 Stockholm, Sweden. bato@kth.se
Veide
Andres
A
Charoenrat
Theppanya
T
Teeri
Tuula
T
Enfors
Sven-Olof
SO
D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D007218:Industrial Microbiology / Q000379:methods*; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism*
D011994:Recombinant Proteins