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  • Chemistry  (1)
  • anaplerotic carbon metabolites  (1)
  • ELECTRONICS AND ELECTRICAL ENGINEERING
  • 1995-1999  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Photosynthesis research 50 (1996), S. 133-148 
    ISSN: 1573-5079
    Keywords: anaplerotic carbon metabolites ; dark respiration ; hexose phosphates ; nitrogen-limitation ; orthophosphate ; photosynthesis ; starch ; sucrose
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Prolonged inorganic nitrogen (NO3 −+NH4 +) limitation of non-N2-fixing soybean plants affected leaflet photosynthesis rates, photosynthate accumulation rates and levels, and anaplerotic carbon metabolite levels. Leaflets of nitrogen-limited (N-Lim), 27–31-day-old plants displayed ≈ 15 to 23% lower photosynthesis rates than leaflets of nitrogen-sufficient (N-Suff) plants. In contrast, N-Lim plant leaflets displayed higher sucrose and starch levels and rates of accumulation, as well as higher levels of carbon metabolites associated with sucrose and starch synthesis, e. g., glycerate-3-phosphate and glucose phosphates, than N-Suff plant leaflets. Concurrently, levels of soluble protein, chlorophyll, and anaplerotic metabolites, e.g., malate and phosphoenolpyruvate, were lower in leaflets of N-Lim plants than N-Suff plants, suggesting that the enzymes of the anaplerotic carbon metabolite pathway were lower in activity in N-Lim plant leaflets. Malate net accumulation rates in the earliest part of the illumination period were lower in N-Lim than in N-Suff plant leaflets; however, by the midday period, malate accumulation rate in N-Lim plant leaflets exceeded that in leaflets of N-Suff plants. Further, soluble protein accumulation rates in leaflets of N-Suff and N-Lim plants were similar, and the rate of dark respiration, measured in the early part of the dark period, was higher in N-Lim plant leaflets than in N-Suff plant leaflets. It was concluded that during prolonged N-limitation, foliar metabolite conditions favored the channelling of a large proportion of the carbon assimilate into sucrose and starch, while assimilate flow through the anaplerotic pathway was diminished. However, in some daytime periods, there was a normal level of carbon assimilate channelled through the anaplerotic pathway for ultimate use in amino acid and protein synthesis.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0006-3592
    Keywords: methanol sensor ; methanol monitoring and control ; methylotrophic yeast fermentation ; Pichia pastoris ; transferrin ; shake-flask cultures ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The methylotrophic yeast Pichia pastoris can be used to express recombinant genes at high levels under the control of the methanol-inducible alcohol oxidase 1 (AOX1) promoter. Accurate regulation of the methanol concentration in P. pastoris cultures is necessary to maintain induction, while preventing accumulation of methanol to cytotoxic levels. We developed an inexpensive methanol sensor that uses a gas-permeable silicone rubber tube immersed in the culture medium and an organic solvent vapor detector. The sensor was used to monitor methanol concentration continuously throughout a fed-batch shake-flask culture of a P. pastoris clone producing the N-lobe of human transferrin. The sensor calibration was stable for the duration of the culture and the output signal accurately reflected the methanol concentration determined off-line by HPLC. A closed-loop control system utilizing this sensor was developed and used to maintain a 0.3% (v/v) methanol concentration in the culture. Use of this system resulted in a fivefold increase in volumetric protein productivity over levels obtained using the conventional fed-batch protocol. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 279-286, 1997.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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