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Kamarthapu V, Ragampeta S, Rao KV, et al. Engineered Pichia pastoris for enhanced production of S-adenosylmethionine. AMB Express. 2013;3:40doi: 10.1186/2191-0855-3-40.
Kamarthapu, V., Ragampeta, S., Rao, K. V., & Reddy, V. D. (2013). Engineered Pichia pastoris for enhanced production of S-adenosylmethionine. AMB Express, 340. https://doi.org/10.1186/2191-0855-3-40
Kamarthapu, Venu, et al. "Engineered Pichia pastoris for enhanced production of S-adenosylmethionine." AMB Express vol. 3 (2013): 40. doi: https://doi.org/10.1186/2191-0855-3-40
Kamarthapu V, Ragampeta S, Rao KV, Reddy VD. Engineered Pichia pastoris for enhanced production of S-adenosylmethionine. AMB Express. 2013 Jul 27;3:40. doi: 10.1186/2191-0855-3-40. eCollection 2013. PMID: 23890127; PMCID: PMC3750815.
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AMB Express. 2013 Jul 27;3:40. doi: 10.1186/2191-0855-3-40. eCollection 2013.

Engineered Pichia pastoris for enhanced production of S-adenosylmethionine.

AMB Express

Venu Kamarthapu, Srinivas Ragampeta, Khareedu Venkateswara Rao, Vudem Dashavantha Reddy

Affiliations

  1. Centre for Plant Molecular Biology, Osmania University, Hyderabad 500 007, India.
  2. National Centre for Mass Spectroscopy, Indian Institute of Chemical Technology, Hyderabad 500 007, India.

PMID: 23890127 PMCID: PMC3750815 DOI: 10.1186/2191-0855-3-40

Abstract

A genetically engineered strain of Pichia pastoris expressing S-adenosylmethionine synthetase gene from Saccharomyces cerevisiae under the control of AOX 1 promoter was developed. Induction of recombinant strain with 1% methanol resulted in the expression of SAM2 protein of ~ 42 kDa, whereas control GS115 showed no such band. Further, the recombinant strain showed 17-fold higher enzyme activity over control. Shake flask cultivation of engineered P. pastoris in BMGY medium supplemented with 1% L-methionine yielded 28 g/L wet cell weight and 0.6 g/L S-adenosylmethionine, whereas control (transformants with vector alone) with similar wet cell weight under identical conditions accumulated 0.018 g/L. The clone cultured in the bioreactor containing enriched methionine medium showed increased WCW (117 g/L) as compared to shake flask cultures and yielded 2.4 g/L S-adenosylmethionine. In spite of expression of SAM 2 gene up to 90 h, S-adenosylmethionine accumulation tended to plateau after 72 h, presumably because of the limited ATP available in the cells at stationery phase. The recombinant P pastoris seems promising as potential source for industrial production of S-adenosylmethionine.

Keywords: Bioreactor; Heterologous host; Pichia pastoris; S-adenosylmethionine synthetase

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