Open AccessProperties of the Hansenula polymorpha ‐derived constitutive GAP promoter, assessed using an HSA reporter gene
Author(s) -
Heo JooHyung,
Hong Won Kyoung,
Cho Eun Young,
Kim Moo Woong,
Kim JeongYoon,
Kim Chul Ho,
Rhee Sang Ki,
Kang Hyun Ah
Publication year - 2003
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1016/s1567-1356(03)00150-8
Subject(s) - biology , expression vector , microbiology and biotechnology , complementary dna , gene expression , glycerol , promoter , human serum albumin , reporter gene , cell culture , gene , biochemistry , recombinant dna , genetics
The glyceraldehyde‐3‐phosphate dehydrogenase promoter, P GAP , was employed to direct the constitutive expression of recombinant human serum albumin (HSA) in Hansenula polymorpha . A set of integration vectors containing the HSA cDNA under the control of P GAP was constructed and the elemental parameters affecting the expression of HSA from P GAP were analyzed. The presence of a 5′‐untranslated region derived from the HSA cDNA and the integration of the expression vector into the GAP locus were shown to improve the expression of HSA under P GAP . Glycerol supported a higher level of HSA expression from P GAP along with a higher cell density than either glucose or methanol. The growth at high glycerol concentrations up to 12% did not cause any significant repression of the cell growth. A high cell density culture, up to 83 g l −1 dry cell weight with a HSA production of 550 mg l −1 , was obtained in less than 32 h of cultivation in a fed‐batch fermentation employing intermittent feeding with 12% glycerol. The GAP promoter‐based HSA expression system showed a higher specific production rate and required a much simpler fermentation process than the MOX promoter‐based system, demonstrating that P GAP can be a practical alternative of the MOX promoter in the large‐scale production of HSA from H. polymorpha .