Abstract
The recombinant Pichia pastoris harboring an improved methionine adenosyltransferase (MAT) shuffled gene was employed to biosynthesize S-adenosyl-l-methionine (SAM). Two l-methionine (l-Met) addition strategies were used to supply the precursor: the batch addition strategy (l-Met was added separately at three time points) and the continuous feeding strategies (l-Met was fed continuously at the rate of 0.1, 0.2, and 0.5 g l−1 h−1, respectively). SAM accumulation, l-Met conversion rate, and SAM productivity with the continuous feeding strategies were all improved over the batch addition strategy, which reached 8.46 ± 0.31 g l−1, 41.7 ± 1.4%, and 0.18 ± 0.01 g l−1 h−1 with the best continuous feeding strategy (0.2 g l−1 h−1), respectively. The bottleneck for SAM production with the low l-Met feeding rate (0.1 g L−1 h−1) was the insufficient l-Met supply. The analysis of the key enzyme activities indicated that the tricarboxylic acid cycle and glycolytic pathway were reduced with the increasing l-Met feeding rate, which decreased the adenosine triphosphate (ATP) synthesis. The MAT activity also decreased as the l-Met feeding rate rose. The reduced ATP synthesis and MAT activity were probably the reason for the low SAM accumulation when the l-Met feeding rate reached 0.5 g l−1 h−1.
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Acknowledgement
We are grateful to the National Basic Research Program (973 program no. 2007CB714306), the National High-Technology Research and Development Program (863 Program 2007AA100601), and the Science and Technology Commission of Shanghai Municipality (07pj14027) for their financial supports to this research.
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Hu, H., Qian, J., Chu, J. et al. Optimization of l-methionine feeding strategy for improving S-adenosyl-l-methionine production by methionine adenosyltransferase overexpressed Pichia pastoris . Appl Microbiol Biotechnol 83, 1105–1114 (2009). https://doi.org/10.1007/s00253-009-1975-y
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DOI: https://doi.org/10.1007/s00253-009-1975-y