title: Engineered Pichia pastoris for enhanced production of S-adenosylmethionine.
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.
authors.affiliation: Centre for Plant Molecular Biology, Osmania University, Hyderabad 500 007, India.
authors.forename: Venu
authors.initials: V
authors.lastname: Kamarthapu
authors.affiliation: National Centre for Mass Spectroscopy, Indian Institute of Chemical Technology, Hyderabad 500 007, India.
authors.forename: Srinivas
authors.initials: S
authors.lastname: Ragampeta
authors.affiliation: Centre for Plant Molecular Biology, Osmania University, Hyderabad 500 007, India.
authors.forename: Khareedu Venkateswara
authors.initials: KV
authors.lastname: Rao
authors.affiliation: Centre for Plant Molecular Biology, Osmania University, Hyderabad 500 007, India.
authors.forename: Vudem Dashavantha
authors.initials: VD
authors.lastname: Reddy
keywords: Bioreactor; Heterologous host; Pichia pastoris; S-adenosylmethionine synthetase
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