Biomedical subjects
S J Pirt
Publications and source records attributed to S J Pirt.
Maintenance energy: a general model for energy-limited and energy-sufficient growth.
The new model proposed to account for the energy requirement for growth includes both a constant maintenance energy term (m) independent of the specific growth rate and a term (m') which decreases linearly with increase in specific growth rate and becomes zero at the maximum specific growth rate. The available data for testing the model do not deviate significantly from the relations predicted. Consistent values of the maximum growth yield (YG) can be derived, irrespective of whether the cultures are energy limited or energy sufficient. Attention is drawn to the possibility that the constant maintenance energy term may be estimated from the maximum specific growth rate.
Conversion of glucose to fatty acids and methane: roles of two mycoplasmal agents.
Two species of obligately anaerobic mycoplasmas were the major components of a methanogenic glucose-limited enrichment culture. In pure culture, one of these organisms, tentatively named Anaeroplasma sp. strain London, was shown to be responsible for the fermentation of glucose to fatty acids, hydrogen, and carbon dioxide; the other mycoplasma was shown to produce methane from hydrogen and carbon dioxide and was named Methanoplasma elizabethii. This same methanogenic mycoplasma contained a low-molecular-weight fluorescent cofactor which had a maximum light absorbance at 430 nm. When both species of mycoplasmas were grown together on glucose, fermentation products included fatty acids and methane. For the first time, mycoplasmas are implicated as agents of anaerobic degradation and methanogenesis in a sewage sludge digester.
The effects of oxygen and carbon dioxide partial pressures on the rate and efficiency of algal (Chlorella) photosynthesis.
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Growth of human diploid cells (strain MRC-5) in defined medium; replacement of serum by a fraction of serum ultrafiltrate.
A calf serum ultrafiltrate fraction permitted growth for at least 3.5 generations, including one subculture, of MRC-5 cells in defined medium in the absence of whole serum. The active material has a molecular weight of 10 000 Daltons or less. This suggests that there may be no requirement for a large macromolecular component of serum. The ultrafiltrate was assayed by maximum cell yield from a serum-limited inoculum in a defined medium containing non-limiting amounts of vitamins, amino acids, glucose, a 68-component supplement, iron and methylcellulose. The levels of vitamins, amino acids and glucose were based on quantitative measurements of uptake and the levels of the other components by minimum amount required for maximum yield in defined medium without ultrafiltrate or serum. With excess ultrafiltrate maximum cell yield was limited by the defined part of the medium, probably the supplement. The cell doubling time in defined medium with ultrafiltrate fractions was 70 h compared with 27 h in the medium with serum. Excess ultrafiltrate did not inhibit growth. The lowered growth rate is attributed to a nutritional deficiency in the supplement.
Biomass yields of Chlorella from iron (Yx/Fe) in iron-limited batch cultures.
The maximum biomass in iron-limited photosynthetic batch cultures of chlorella increased as the logarithm of the iron concentration. The growth yield from iron (UxFe) showed a marked inverse relation to the specific growth rate. The maximum biomass yield, g dry biomass/g iron consumed, was 7.5 X 10(3) with specific growth rate 0.108h-1; the minimum was 0.79 X 10(3) with specific growth rate 0.145 h-1. The maximum specific growth rate in the exponential phase of Fe limited cultures varied as the initial Fe concentration. Fe-limited growth made the cells adhere to a glass surface.
Development of the agar disk method for the rapid selection of cephalosporin producers with improved yields.
To screen the abilities of mutant strains of Cephalosporium to produce cephalosporin C, colonies of the organism were grown on the surface of small (4-mm diameter) disks of agar medium. After incubation of the disks for periods of up to 5 days, the antibiotic contents of the disks were assayed by placing them on agar plates of the assay organism and determining the diameters of the inhibition zones. The amount of nitrogen source in the agar disk medium was used to control the amount of antibiotic produced in the disk and, thus, the sensitivity of screening. The relation of agar disk inhibition zone diameters to log shake-flask titers was linear with short incubation times (2 to 3 days) of the disks, but shifted towards a higher order with prolonged incubation (4 to 5 days). The optimum incubation time for the disks was 4 to 5 days, and then a 15% difference in zone diameters was significant with 10 disks per sample. The minimum difference between the shake-flask titers, which could be detected by the agar disk method with 10 disks per sample, was about 30% with 5 days of incubation for the disks. The results suggest that the shake-flask culture underestimated the degree of improvement in strain productivity.
Nutrition and factors limiting the growth of a methanogenic bacterium (Methanobacterium thermoautotrophicum).
The purification of Methanobacterium thermoautotrophicum from a culture contaminated with a heterotrophic organism is described. A defined inorganic medium under H2/CO2 (80:20 v/v) has been developed to support growth of M. thermoautotrophicum up to a concentration of at least 1.7 g dry weight/l. In a conventional medium iron and nitrogen sources were found to be growth-limiting factors. Throughout most of the culture period the rate of transfer of hydrogen or carbon dioxide from gas to liquid was the factor which controlled the growth rate. The growth yields of bacteria were in the range of 0.6-1.6 g dry weight/mole CH4.
Growth dynamics of Agrobacterium tumefaciens in chemostat cultures limited by carbon source and mineral nutrients.
Agrobacterium tumefaciens was grown in a chemostat in a chemically-defined medium which hs alpha-methyl D-glucoside, magnesium, manganese, phosphate or urea as the growth-limiting nutrient. Steady-state biomass concentrations were dependent on the specific growth rate of the organism when alpha-methyl D-glucoside, manganese or phosphate were growth-limiting nutrients. During magnesium-limited growth, large undamped oscillations in biomass concentration occurred. In all chemostat cultures a variant organism was selected which had lost the ability to grow in the medium supplied, bur survived on products of carbon metabolism derived from the wild-type.
Protein turnover measured by 18-O exchange with H2-18O in non-growing cells of Agrobacterium tumefaciens.
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Factors influencing the formation and stability of D-glucoside 3-dehydrogenase activity in cultures of Agrobacterium tumefaciens.
D-glucoside 3-dehydrogenase specific activity in Agrobacterium tumefaciens was maximal towards the end of the exponential growth phase of batch cultures; over 90% of the activity disappeared within the next 15 h. Manganese ions, although essential for growth of the organism, strongly repressed D-glucoside 3-dehydrogenase synthesis in sucrose medium but had little effect when the carbon source was methyl alpha-D-glucoside. D-Glucoside 3-dehydrogenase activity increased linearly with increasing specific growth rate in chemostat cultures limited by carbon, nitrogen, phosphate or manganese when methyl alpha-D-glucoside was the carbon source. High enzyme activity was found with sucrose as carbon source only when the growth medium was manganese-limited. D-Glucoside 3-dehydrogenase activity disappeared from A. tumefaciens incubated in carbon- and nitrogen-free medium or in nitrogen-free medium containing succinate, but on continued incubation the activity returned and was then stable. The recovery of activity could be prevented by chloramphenicol or erythromycin. Bacteria containing the recovered dehydrogenase activity could not convert sucrose to 3-ketosucrose when oxygen acted as the terminal electron acceptor, but produced 3-ketosucrose at the normal rate in the presence of ferricyanide. D-Glucoside 3-dehydrogenase activity disappeared irreversibly from bacteria incubated in nitrogen-free medium containing sucrose. Loss of activity followed first order kinetics in bacteria taken from nitrogen-, phosphate- or manganese-limited chemostat steady states; an accelerating rate of decay occurred in cells grown under carbon-limitation. 8-Hydroxyquinoline, chloramphenicol, erythromycin, 2,4-dinitrophenol and manganese ions could reduce the rate of decay.
The quantitative requirements of human diploid cells (strain MRC-5) for amino acids, vitamins and serum.
The uptakes of all essential amino acids, vitamins (except riboflavin), glucose and serum during growth of human diploid cells (MRC-5) were determined. The amino acid uptakes varied considerably with the conditions of culture. The glucose requirement is several times greater than that for mouse LS or human HeLa cells. These analytical results were used to modify the medium so as to ensure that an excess of all defined medium constituents was present and pH was not limiting during study of the serum requirements. It was then found that maximum cell populations were directly proportional to the serum concentration. Hence the growth was limited by the supply of an unknown growth factor in serum. The serum growth factor was not replaced by a mixture of over 60 vitamins, co-enzymes, hormones and other organic and inorganic compounds considered to be possible growth factors, although this mixture did not lower the growth rate and somewhat (22%) increased the yield from the serum growth factor. The unit of serum growth factor is precisely defined in terms of the amount in a standard batch of calf serum. This standard contains 10 units/ml whereas the other batch of serum used contained only 5 units/ml.
The production of interferon by chemostat cultures of mouse LS-cells grown in chemically-defined, protein-free medium.
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Penicillin production by lysine auxotrophs of Penicillium chrysogenum.
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Inhibition by 3-deoxy-3-fluoro-D-glucose of the utilization of lactose and other carbon sources by Escherichia coli.
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Possible adverse effect of protozoa on effluent purification systems.
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Inhibition of the utilization of lactose and other carbon sources by 3-deoxy-3-fluoro-D-glucose in Escherichia coli.
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Induction of mouse interferon in a chemically defined system.
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