Use of different carbon sources in cultivation of recombinant Pichia pastoris for angiostatin production |
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| Affiliation: | 1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;2. Institute of Biochemistry and Cell Biology, Shanghai Institute of Biological Science, Chinese Academy of Science, Shanghai 200031, China;1. Institute of Biochemical Engineering, Technische Universität München, Boltzmannstr. 15, 85748 Garching, Germany;2. W42 Industrial Biotechnology GmbH, Otto-Hahn-Str. 15, 44227 Dortmund, Germany;3. W42 Consulting GmbH, Südstr. 19, 82377 Penzberg, Germany;1. Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam 781039, India;2. Advanced Centre for Biochemical Engineering, University College London, WC1E 6BT, United Kingdom;1. Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen Germany;2. Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Giessen Germany;3. Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Giessen Germany;4. Department of Chemical Engineering, Kansas State University, Manhattan USA |
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| Abstract: | To improve the growth of recombinant Pichia pastoris with a phenotype of MutS and expression of angiostatin, the effects of glycerol, sorbitol, acetate and lactic acid which were, respectively, added together with methanol in the expression phase, were studied in a 5-l fermentor. Methanol concentration was automatically controlled at 5 g/l by a methanol monitor and control system, while the feeding of the other carbon source was manually adjusted. The angiostatin production level was 108 mg/l when glycerol was added at an initial rate of 2.3 g/h and gradually increased to 9.9 g/h within an induction period of 96 h. The angiostatin concentration was 141 mg/l as sorbitol was used, while only 52 mg/l were obtained on acetate. The highest angiostatin production of 191 mg/l was achieved as lactic acid was used; whose feeding rate was gradually increased from 2.6 to 11.3 g/h. Lactic acid accumulated during the induction phase and reached 6.3 g/l at the end of fermentation. However, the accumulation of lactic acid did not interfere with angiostatin production, indicating that lactic acid to be a non-repressive carbon source. The average productivity and specific productivity of angiostatin obtained on lactic acid and methanol were, respectively, 2.96 and 0.044 mg/(g h), 1.7- and 2.5-fold of those obtained in the fermentation fed with glycerol and methanol. |
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