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Open AccessArticle

Transcriptomic Analysis of the Influence of Methanol Assimilation on the Gene Expression in the Recombinant Pichia pastoris Producing Hirudin Variant 3

1
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China
2
College of Basic Science, Tianjin Agricultural University, Tianjin 300384, China
*
Author to whom correspondence should be addressed.
Genes 2019, 10(8), 606; https://doi.org/10.3390/genes10080606
Received: 27 July 2019 / Accepted: 7 August 2019 / Published: 12 August 2019
(This article belongs to the Section Microbial Genetics and Genomics)
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Abstract

Hirudin and its variants, as strong inhibitors against thrombin, are present in the saliva of leeches and are recognized as potent anticoagulants. However, their yield is far from the clinical requirement up to now. In this study, the production of hirudin variant 3 (HV3) was successfully realized by cultivating the recombinant Pichia pastoris GS115/pPIC9K-hv3 under the regulation of the promoter of AOX1 encoding alcohol oxidase (AOX). The antithrombin activity in the fermentation broth reached the maximum value of 5000 ATU/mL. To explore an effective strategy for improving HV3 production in the future, we investigated the influence of methanol assimilation on the general gene expression in this recombinant by transcriptomic study. The results showed that methanol was partially oxidized into CO2, and the rest was converted into glycerone-P which subsequently entered into central carbon metabolism, energy metabolism, and amino acid biosynthesis. However, the later metabolic processes were almost all down-regulated. Therefore, we propose that the up-regulated central carbon metabolism, energy, and amino acid metabolism should be beneficial for methanol assimilation, which would accordingly improve the production of HV3. View Full-Text
Keywords: anticoagulant; methanol induction; fermentation; hirudin; Pichia pastoris anticoagulant; methanol induction; fermentation; hirudin; Pichia pastoris
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Li, T.; Ma, J.; Xu, Z.; Wang, S.; Wang, N.; Shao, S.; Yang, W.; Huang, L.; Liu, Y. Transcriptomic Analysis of the Influence of Methanol Assimilation on the Gene Expression in the Recombinant Pichia pastoris Producing Hirudin Variant 3. Genes 2019, 10, 606.

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