Oxygen vectors used for S-adenosylmethionine production in recombinant Pichia pastoris with sorbitol as supplemental carbon source

Zhang, Jianguo; Wang, Xuedong; Zhang, Ji-Ning; Wei, Dongzhi

doi:10.1263/jbb.105.335

Cited by 33 publications

(19 citation statements)

References 24 publications

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“…In recent studies on the co‐production of SAM and GSH by either Saccharomyces cerevisiae or Candida utilis , screening of strains with high performance and optimization of fermentation conditions were the commonly used approaches to obtain high concentrations of these sulfur‐containing compounds . It is generally known that SAM and GSH are biosynthesized with the consumption of ATP in yeast cells, and that intracellular ATP level and ATP regeneration can act as the limiting factors for efficient co‐production of SAM and GSH . According to our previous studies, enhanced co‐production of SAM and GSH was achieved by either using an ATP‐oriented amino acid addition strategy or the addition of citrate as an auxiliary energy substrate to the fermentation medium .…”

Section: Introductionmentioning

confidence: 99%

“…The intracellular ATP supply and consumption can be regulated by controlling the DO levels during batch fermentation processes. An appropriate DO level will favor the increase in ATP generation, which in turn will facilitate the overproduction of SAM and GSH . However, when the DO is above a certain level, the reducing molecule GSH will be oxidized to GSSG, which will inversely decrease the co‐production of SAM and GSH .…”

Section: Introductionmentioning

confidence: 99%

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Efficient co‐production of S‐adenosylmethionine and glutathione by Candida utilis: effect of dissolved oxygen on enzyme activity and energy supply

Wang

Wei

2017

J of Chemical Tech & Biotech

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BACKGROUND As both S‐adenosylmethionine (SAM) and glutathione (GSH) are biosynthesized with ATP consumption in yeast cells, their efficient co‐production not only depends on high activities of key enzymes, but also on sufficient energy supply. In this study, dissolved oxygen (DO) was optimized to improve enzyme activity and energy supply for high co‐production of SAM and GSH. RESULTS Batch culture of Candida utilis CCTCC M 209298 under different DO control modes was carried out in a 5 L stirred fermentor. Based on the analyses of kinetic parameters and metabolic fluxes of key nodes involved in the metabolism of SAM and GSH, a two‐stage DO control strategy with an initial agitation rate of 350 rpm followed by a constant DO level of 30% was designed for the overproduction of SAM and GSH. A maximum co‐production of SAM and GSH of 593.9 mg L−1 was achieved using the two‐stage DO control strategy, which was much higher than that obtained from other DO control modes. CONCLUSION Physiological mechanism analysis revealed that the improved co‐production of SAM and GSH was closely related to the increase in key enzyme activities and sufficient energy supply under the two‐stage DO control strategy. © 2017 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Efficient co‐production of S‐adenosylmethionine and glutathione by Candida utilis: effect of dissolved oxygen on enzyme activity and energy supply

Wang

Wei

2017

J of Chemical Tech & Biotech

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“…SAM2 gene was expressed and cystathione-β-synthase gene was knocked out from P. pastoris for production of SAM (He et al 2006; Yu and Shen 2012). Enhanced accumulation of SAM was achieved by manipulating the culture conditions like increased oxygen levels and nitrogen source (Chen et al 2007; Zhang et al 2008a; Zhang et al 2008b; Chu et al 2013). Earlier studies reported elevated intracellular production of SAM using recombinant P. pastoris; however, these reports did not confirm the expression of the enzyme by SDS-PAGE analyses, and molecular characterization of the accumulated SAM using MS/MS method.…”

Section: Introductionmentioning

confidence: 99%

Engineered Pichia pastoris for enhanced production of S-adenosylmethionine

et al. 2013

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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.

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“…Organic compounds, such as hydrocarbons [20], fluorocarbons [5], and soybean oil [9], can be used as oxygen vectors to intensify oxygen supply. Another technology (water-in-oil fermentation technology) has been developed to enhance the performance of highly viscous fermentation processes [12].…”

Section: Introductionmentioning

confidence: 99%

Hydrogen peroxide (H2O2) supply significantly improves xanthan gum production mediated by Xanthomonas campestris in vitro

Cheng

Lin

Zhang

2012

Journal of Industrial Microbiology and Biotechnology

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To improve xanthan gum productivity, a strategy of adding hydrogen peroxide (H₂O₂) was studied. The method could intensify oxygen supply through degradation of H₂O₂ to oxygen (O₂). In shake flask testing, the xanthan gum yield reached 2.8% (improved by 39.4%) when adding 12.5 mM H₂O₂ after 24 h of fermentation. In fermentor testing, it was obvious that the oxygen conditions varied with the H₂O₂ addition time. Eventually, gum yield of 4.2% (w/w) was achieved (increased by 27.3%). Compared with the method of intense mixing and increasing the air flow rate, adding H₂O₂ to improve the dissolved oxygen concentration was more effective and much better. Moreover, addition of H₂O₂ improved the quality of xanthan gum; the pyruvate content of xanthan was 4.4% (w/w), higher than that of the control (3.2%).

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Oxygen vectors used for S-adenosylmethionine production in recombinant Pichia pastoris with sorbitol as supplemental carbon source

Cited by 33 publications

References 24 publications

Efficient co‐production of S‐adenosylmethionine and glutathione by Candida utilis: effect of dissolved oxygen on enzyme activity and energy supply

Efficient co‐production of S‐adenosylmethionine and glutathione by Candida utilis: effect of dissolved oxygen on enzyme activity and energy supply

Engineered Pichia pastoris for enhanced production of S-adenosylmethionine

Hydrogen peroxide (H2O2) supply significantly improves xanthan gum production mediated by Xanthomonas campestris in vitro

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