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Independent exponential feeding of glycerol and methanol for fed-batch culture of recombinant Hansenula polymorpha DL-1

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Abstract

As a novel feeding strategy for aptomizing human epidermal growth factor (hEGF) production with a recombinant Hansenula polymorpha DL-1 using the methanol oxidase (MOX) promoter in H. polymorpha DL-1, independent exponential feeding of two substrates was used. A simple kinetic model considering the cell growth on two substrates was established and used to calculate the respective feeding rates of glycerol and methanol. In the fedbatch culture with methanol-only feeding, the optimal set point of specific growth rate on methanol was found to be 0.10 h−1. When the fed-batch cultures were conducted by the independent feeding of glycerol and methanol, the actual specific growth rate on glycerol and methanol was slightly lower than the set point of specific growth rate. By the uncoupled feeding of glycerol and methanol the volumetric productivity of hEGF increased from 6.4 to 8.0 mg/(L·h), compared with methanol-only feeding.

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Abbreviations

D 0 :

feed glucose concentration (g/L)

F G :

feeding rate of glycerol (L/h)

F M :

feeding rate of methanol (L/h)

G :

glycerol concentration (g/L)

G 0 :

feed glycerol concentration (g/L)

M :

methanol concentration (g/L)

M 0 :

feed methanol concentration (g/L)

P :

hEGF concentration (mg/L)

P max :

hEGF concentration at end of culture (mg/L)

q G :

specific glycerol consumption rate (g/[g·h])

q M :

specific methanol consumption rate (g/[g·h])

t :

time (h)

t f :

final culture time (h)

V :

culture volume (L)

V 0 :

culture volume at beginning of fed-batch phase (L)

X :

cell concentration (g/L)

X 0 :

cell concentration at beginning of fed-batch phase (g/L)

X max :

cell concentration at end of culture (g/L)

Y P/X :

product yield on cell mass (mg/g)

Y X/D :

cell yield on glucose (g/g)

Y X/G :

cell yield on glycerol (g/g)

Y X/M :

cell yield on methanol (g/g)

μ act :

actual specific growth rate (h−1)

μ G :

specific growth rate on glycerol (h−1)

μ Glu,sp :

set point of specific growth rate on glucose (h−1)

μ G,sp :

set point of specific growth rate on glycerol (h−1)

μ M :

specific growth rate on methanol (h−1)

μ M,sp :

set point of specific growth rate on methanol (h−1)

π:

specific production rate (mg/[g·h])

Π:

volumetric productivity (mg/[L·h])

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Authors and Affiliations

  1. Graduate School of Biotechnology, Korea University, 136-701, Soul, Korea

    H. Moon & I. H. Kim

  2. Department of Chemical and Biochemical Engineering, Korea University, 136-701, Seoul, Korea

    S. W. Kim, J. Lee & S. I. Hong

  3. Korea Research Institute of Biosience and Biotechnology, 305-600, Taejon, Korea

    S. K. Rhee, E. S. Choi & H. A. Kang

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  1. H. Moon
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  2. S. W. Kim
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  4. S. K. Rhee
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Correspondence to S. I. Hong.

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Moon, H., Kim, S.W., Lee, J. et al. Independent exponential feeding of glycerol and methanol for fed-batch culture of recombinant Hansenula polymorpha DL-1. Appl Biochem Biotechnol 111, 65–79 (2003). https://doi.org/10.1385/ABAB:111:2:65

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  • DOI: https://doi.org/10.1385/ABAB:111:2:65

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