Br J Nutr. "Citation Classic": Improvement of phosphorus availability by microbial phytase in broilers and pigs. Br J Nutr. 1990 Sep.
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Biomedical subjects
Publications and source records attributed to R F Beudeker.
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Techniques have been developed to produce microbial phytase for addition to diets for simple-stomached animals, with the aim to improve phosphorus availability from phytate-P in plant sources. The activity of the crude microbial phytase showed pH optima at pH 5.5 and 2.5. The enzyme was able to degrade phytate in vitro in soya-bean meal, maize and a liquid compound feed for pigs. When microbial phytase was added to low-P diets for broilers the availability of P increased to over 60% and the amount of P in the droppings decreased by 50%. The growth rate and feed conversion ratio on the low-P diets containing microbial phytase were comparable to or even better than those obtained on control diets. Addition of microbial phytase to diets for growing pigs increased the apparent absorbability of P by 24%. The amount of P in the faeces was 35% lower.
Ion-exchange chromatography of extracts derived from Chlorella sorokiniana mutant strain (oxygen resistant) yielded two separate activity peaks of glutamine synthetase (GS). GS(I) and GS(II) were purified 220- and 187-fold and have molecular weights of approximately 398,000 and 360,000, respectively. Both enzymes are composed of eight identical subunits with a subunit molecular weight of 47,000 for GS(I) and 43,000 for GS(II). The amino acid composition, catalytic, and immunological properties for both enzymes are similar.
The utilization of nitrate and ammonia as nitrogen sources had different effects on the metabolism of glycolate in Cholorella sorokiniana. During photolithotrophic growth with nitrate as nitrogen source, glycolate was metabolized via the glycine-serine pathway. Ammonia, produced as a result of glycolate metabolism, was reassimilated by glutamine synthetase. Two isoforms of this enzyme were present at different relative abundance in C. sorokiniana wild type and in a mutant with an increased capacity for the metabolism of glycolate (strain OR).During photolithotrophic growth in the presence of ammonia as sole nitrogen source, several lines of evidence indicated that glycolate was metabolized to malate, pyruvate, tricarboxylic acid cycle intermediates and related amino acids in C. sorokiniana wild-type cells. Malate synthase was induced and glycine decarboxylase and serine-glyoxylate aminotransferase were repressed in cells grown with ammonia. An inverse correlation was observed between aminating NADPH-glutamate dehydrogenase and the in vivo glycine decarboxylation rate.
Under a gas atmosphere of 99% O2/1% CO2, wild-type cells of Chlorella sorokiniana excreted 12% of their dry weight as glycolate during photolithotrophic growth, whereas mutant cells excreted glycolate at only 3% of the cellular dry weight. The observed difference in glycolate excretion by the two cell types appears to be due to a different capacity for the metabolism of glycolate, rather than to a different glycolate formation rate. This was concluded from experiments in which the metabolism of glycolate via the glycine-serine pathway was inhibited by the addition of isoniazid. Under such conditions, glycolate excretion rates for both cell types were identical. The mutant appeared to have significantly higher specific activities of glycine decarboxylase, serine hydroxymethyltransferase, serine-glyoxylate aminotransferase, glycerate kinase, and phosphoglycolate phosphatase than did the wild type. The specific activities of D-ribulose-1,5-bisphosphate carboxylase/oxygenase, glycolate dehydrogenase, glyoxylate-aminotransferase, and hydroxypyruvate reductase were the same for wild-type and mutant cells. The internal pool sizes of ammonia and amino acids increased in wild-type cells grown under high-oxygen concentrations but were hardly affected by high oxygen tensions in the mutant cells. Our results indicate that, under the growth conditions applied, the decarboxylation of glycine becomes the rate-limiting step of the glycine-serine pathway for the wild-type cells of C. sorokiniana.
Recent studies on the ecophysiology of the obligate chemolithotroph Thiobacillus neapolitanus have given better insight into its specialization for an autotrophic mode of life. This appears not only from its high constitutive levels of autotrophic enzymes, but also from its possession of carboxysomes, which seem to be specialized organelles for CO2 fixation and concentrating reducing power. At the same time, these organisms are metabolically versatile with respect to nitrogen assimilation pathways, and during starvation are able to utilize endogenous resources such as polyglucose for carbon and energy. Studies on the facultative chemolithotrophs such as Thiobacillus novellus and Thiobacillus A2 have shown that they can grow mixotrophically on mixtures of inorganic and organic substrates, i.e. they can utilize these compounds simultaneously provided that they are growth limiting. Thiobacillus A2 displays a remarkable flexibility not only with respect to the organic substrates that it can utilize but, for example, also in the choice of various pathways for glucose metabolism. Competition experiments carried out between specialized and versatile thiobacilli strongly indicate that the ecological advantage of the versatile thiobacilli may lie not so much in their short-term flexibility, but rather in their ability to grow mixotrophically. Studies on most heterotrophic chemolithotrophs are still in their infancy. Promising progress has been made in the study of the physiology of Beggiatoa species. Renewed interest in the sulphur-oxidizing bacteria stems from recent findings about their role in food chains, and their possible application in industry.
The results of ecophysiological studies on obligately and facultatively chemolithotrophic thiobacilli performed over the past years clearly show that the two types of organisms occupy different ecological niches. Chemostat experiments with cultures of the obligate chemolithotroph Thiobacillus neapolitanus and the facultative chemolithotroph Thiobacillus A2 have been carried out to explain the competitiveness of T. neapolitanus under conditions of strongly fluctuating substrate supply. Thiobacillus neapolitanus appeared to be very resistant to starvation periods whereafter it could oxidize sulfide (or thiosulfate) almost instantaneously at the original rate. Under alternate supply of 4 h sulfide and 4 h sulfate (or acetate which does not support growth of the organism either) to a chemostat culture of T. neapolitanus (D=0.05h-1) the sulfide concentration in the growth vessel never reached levels higher than 4 micrometers. This strategy is aimed at maximal reactivity. In contrast to T. neapolitanus the facultative chemolithotroph T.A2 appeared to be very flexible with respect to its energy generation. Under alternate supply of 4 h sulfide and 4 acetate (D=0.05h-1) T.A2 was able to grow continuously since it directed its metabolism to either heterotrophy or autotrophy by rapid induction-repression mechanisms. This flexible strategy seems to be incompatible with a reactive strategy within one organism, since the oxidation capacity for sulfide decreased during the acetate period resulting in accumulation of sulfide during the sulfide period. It is concluded that T.A2 needs a continuous supply of an inorganic and an organic substrate to thrive whereas T. neapolitanus needs only a continuous supply of a reduced inorganic sulfur source but also will persist in environments with interrupted addition of sulfide provided that the starvation period does not last too long.