Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reexamination Certificate
2005-01-04
2005-01-04
Ketter, James (Department: 1636)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S023100, C536S023200, C435S004000, C435S006120, C435S041000, C435S069100, C435S071100, C435S320100, C435S255100, C435S256100
Reexamination Certificate
active
06838555
ABSTRACT:
Disclosed areHansenula polymorphamutants useful as host cells through which various proteins can be produced as being intact at high yield and a process for preparing recombinant proteins using the host cells. Using various vectors,Hansenula polymorphais made to be a mutant which is deprived of methanol assimilating ability and incapable of utilizing methanol as a carbon source. ThisHansenula polymorphamutant is used as a high yield host to produce recombinant proteins without continuous feeding of methanol, with the aid of an expression cassette carrying a promoter capable of inducing the expression at a low concentration of methanol. Further, the mutant is also lacking in carboxypeptidase Y, protease Y and/or carboxypeptidase a activity, so the recombinant protein of interest is not degraded at its carboxyl terminal when being expressed in the cell. Thus, intact recombinant protein can be obtained. Also, there is disclosed a pop-out technique in which a recombinant protein expression cassette is inserted into a MOX gene site of the mutant and is allowed to pop out therefrom, thereby utilizing the mutant as a host for general use in producing various proteins of interest.
REFERENCES:
patent: WO 0052133 (2000-08-01), None
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Hodgkins, Martin et al., Expression of the Glucose Oxidase Gene from Aspergillus niger in Hansenula polymorpha and its Use as a Reporter Gene to Isolate Regulatory Mutations, Yeast, vol. 9, pp. 625-635, Jan. 14, 1993.
Hinnen, Albert et al., Gene Expression in Recombinant Yeast, pp. 121-193, Ciba-Geigy AG, Basel, Switzerland.
Romanos, Michael A. et al., Recombinant Bordetella pertussis pertactin (P69) from the yeast Pichia pastoris: high-level production and immunological properties, Vaccine, vol. 9, Dec. 1991, pp. 901-906.
Agaphonov Michael
Bae Junghoon
Choi Euisung
Kang Hyunah
Kim Moowoong
Fox Rothschild LLP
Ketter James
Korea Research Institute of Bioscience & Biotechnology
Lambertson David A.
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