Distribution of cellular amino acids, protein, and total organic nitrogen during fruit body development in Flammulina velutipes. II. Growth on potato–glucose solution

1982 ◽  
Vol 60 (8) ◽  
pp. 1342-1351 ◽  
Author(s):  
Hans E. Gruen ◽  
William M. Wong

In Flammulina velutipes grown on potato–glucose solution the dry weight of mycelium decreased when fruitbodies elongated rapidly. Loss of dry weight by aborted primordia and stunted fruitbodies paralleled gains by large fruitbodies. These gains exceeded the losses by the rest of the colony and indicate that large fruitbodies derived their substrates both from the medium and from materials stored in the rest of the colony. Close to 43% of the organic nitrogen in the medium was consumed before fruitbodies formed and only 8% remained at the end of their growth. Concentrations of total nitrogen, α-amino nitrogen, and alkali-soluble protein declined in the mycelium during most of the growth of the fruitbodies and then remained constant except for a slight final increase in protein. Concentrations of these fractions also decreased in the small fruitbodies and stipes of large fruitbodies but increased in the pilei. Changes in total amounts of these fractions show that gains by all fruitbodies during elongation exceeded the losses by the mycelium and that part of their nitrogen continues to be derived from the medium. Changes in 18 free and 17 protein amino acids were followed quantitatively in the colony. Free arginine, glutamic acid, ornithine, and alanine predominated in the mycelium and small fruitbodies during early growth of the latter, but arginine and ornithine decreased steeply thereafter. During rapid elongation of the large fruitbodies the concentration of arginine increased strongly in the pilei. The increase was much less in the stipes, but successive changes in the rest of the colony suggest that arginine is translocated into the large pilei from the mycelium and possibly the small fruitbodies. Aspartic and glutamic acids, leucine, and alanine predominated in protein of the whole colony until the early growth of fruitbodies, but protein composition changed during rapid elongation. The proportion of valine increased in the mycelium and it became the most abundant bound amino acid in large stipes. [14C]Leucine was fed in replacement medium and incorporation of the label was determined in nitrogenous fractions of different colony portions at the onset and termination of fruitbody growth. Most of the label transported into fruitbodies was in the ethanol-soluble free amino acids.

1982 ◽  
Vol 60 (8) ◽  
pp. 1330-1341 ◽  
Author(s):  
Hans E. Gruen ◽  
William M. Wong

Dry weights of stipes and pilei of Flammulina velutipes grown on nutrient-supplemented sawdust increased throughout the growth period. The pilei weighed only slightly less than the stipes. Amounts of total organic nitrogen, α-amino nitrogen, and alkali-soluble protein increased in the whole pileus and stipe as the fruitbodies elongated, but the concentrations on a dry weight basis decreased although they were always highest in the pileus. The concentration of alkali-insoluble nitrogen increased in both structures and was highest in the stipe. Concentrations of total nitrogen and protein in surface mycelium did not change significantly when fruitbodies formed but increased markedly when their growth ceased. An average of 8.3 mg dry weight of spores containing about 0.6 mg of nitrogen was released during the life of the fruitbody. Changes in 18 free and 17 protein amino acids were followed quantitatively in the pileus and stipe. Glutamic and aspartic acids and alanine were always among the four predominant free amino acids. Lysine and arginine concentrations remained low in the stipes but increased considerably in the expanding pilei. Ornithine levels increased strongly in the stipe during the early part of rapid elongation but remained almost constant and low in the pilei. Free proline was detected only in traces. Valine became the most abundant protein amino acid during elongation, especially in the stipe. There was very little bound methionine and cystine. In surface mycelium levels of free amino acids were low before fruiting and close to the end of fruitbody growth. Protein amino acids increased during that interval but their proportions remained virtually unchanged and valine was not predominant. The concentration of urea remained very low in both pilei and stipes during their growth.


1987 ◽  
Vol 65 (3) ◽  
pp. 518-525 ◽  
Author(s):  
Ema E. Chao ◽  
Hans E. Gruen

In the mycelium of Flammulina velutipes cultured on potato–glucose solution the concentration of water-soluble protein began to decrease steeply before fruit bodies formed, but the total amount of protein per culture decreased only when fruit bodies began to elongate rapidly. The level remained very low during rapid elongation. Proteinase activity per unit weight of mycelium against milk protein and azoalbumin decreased in parallel with the concentration of mycelial protein. Activity against azure blue – hide powder was low until the onset of rapid fruit-body elongation and reached a maximum later during that phase. Azoalbumin was hydrolyzed more actively than hide powder. Proteolytic activity per gram of mycelium against mycelial protein was almost the same before fruiting as during rapid fruit-body elongation. Specific activities per milligram protein against the three types of nonnative substrates increased in parallel to a maximum late during rapid elongation when protein concentration was lowest. Specific activity against native mycelial protein was also higher in extracts from older mycelia. Proteolysis was optimal near neutral pH and very low at pH 3.2. Tests with group-specific proteinase inhibitors showed that metallo-proteinases predominate in the mycelium. Serine, and to a lesser extent carboxyl and thiol proteinases, were also present. Neither endogenous inhibitors nor activators of proteolysis were detected.


1981 ◽  
Vol 59 (11) ◽  
pp. 2102-2107 ◽  
Author(s):  
Robert C. Evans ◽  
Candace L. Black

The effect of 23 organic and 3 inorganic nitrogen sources on growth, sporulation, and polyphenoloxidase activity was measured in Bipolaris maydis race T incubated with or without a xylose supplement. Mycelial dry weight tends to be highest on acidic and other polar amino acids and least on nonpolar ones; organic nitrogen sources generally result in greater dry weight than inorganic ones. Changes in nitrogen concentration and pH of media influence growth and sporulation, but only pH alters the relative effectiveness of the nitrogen sources on these processes. The addition of xylose to the growth media has little effect on growth. However, in the presence of lysine, serine, asparagine, glycine, γ-alanine, alanine, and α-aminobutyrate, xylose causes an increase in sporulation and a concomitant decrease in polyphenoloxidase activity. There is no consistent pattern resulting from xylose addition in the presence of the other nitrogen sources.


1972 ◽  
Vol 50 (4) ◽  
pp. 803-818 ◽  
Author(s):  
Hans E. Gruen ◽  
Sheue-heng Wu

Isolated Flammulina velutipes fruit bodies were cultured under sterile conditions with the cut base immersed in water or solutions. Stipe elongation on water was only 6% of normal for fruit bodies isolated at 1.1–2.0 cm length, 19% at 5.1–6.0 cm, and the same as for fruit bodies attached to mycelium at 9.1–10.0 cm. Fruit bodies not immersed in water grew less in a saturated atmosphere than those in water. The mycelium must supply other substances than water for normal elongation during about two-thirds of the growth period, and only water thereafter. Isolated fruit bodies fed with filtered glucose, trehalose, sucrose, or mannitol grew better than on water. Maltose and fructose increased elongation only slightly, and sorbose had no effect. Potato extract, yeast extract, and casein hydrolysate gave no or very little growth promotion, but addition of glucose strongly increased growth on the natural extracts compared to glucose alone. Of 21 amino acids added separately to glucose, only asparagine, hydroxyproline, arginine, and to a lesser extent glutamine, stimulated growth of isolated fruit bodies. Growth was not promoted by pure asparagine, glutamine, and serine, or by thiamin or indoleacetic acid. Growth was inhibited by urea, ammonium nitrate, and ammonium tartrate with or without glucose.Growth promoting substances were most effective in young fruit bodies and except for glucose the promotion disappeared in fruit bodies isolated at 6.1–7.0 cm length, which corresponds to the end of the period of rapid elongation. Apical portions of fruit bodies with caps grew better on glucose than whole fruit bodies. Growth of decapitated isolated stipes was not promoted by nutrients.


1995 ◽  
Vol 42 (11) ◽  
pp. 907-912 ◽  
Author(s):  
Takao TERASHITA ◽  
Makoto NAWAMA ◽  
Kentaro YOSHIKAWA ◽  
Jiko SHISHIYAMA

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ramona Wördemann ◽  
Lars Wiefel ◽  
Volker F. Wendisch ◽  
Alexander Steinbüchel

AbstractCyanophycin (multi-l-arginyl-poly-l-aspartic acid; also known as cyanophycin grana peptide [CGP]) is a biopolymer that could be used in various fields, for example, as a potential precursor for the synthesis of polyaspartic acid or for the production of CGP-derived dipeptides. To extend the applications of this polymer, it is therefore of interest to synthesize CGP with different compositions. A recent re-evaluation of the CGP synthesis in C. glutamicum has shown that C. glutamicum is a potentially interesting microorganism for CGP synthesis with a high content of alternative amino acids. This study shows that the amount of alternative amino acids can be increased by using mutants of C. glutamicum with altered amino acid biosynthesis. With the DM1729 mutant, the lysine content in the polymer could be increased up to 33.5 mol%. Furthermore, an ornithine content of up to 12.6 mol% was achieved with ORN2(Pgdh4). How much water-soluble or insoluble CGP is synthesized is strongly related to the used cyanophycin synthetase. CphADh synthesizes soluble CGP exclusively. However, soluble CGP could also be isolated from cells expressing CphA6308Δ1 or CphA6308Δ1_C595S in addition to insoluble CGP in all examined strains. The point mutation in CphA6308Δ1_C595S partially resulted in a higher lysine content. In addition, the CGP content could be increased to 36% of the cell dry weight under optimizing growth conditions in C. glutamicum ATCC13032. All known alternative major amino acids for CGP synthesis (lysine, ornithine, citrulline, and glutamic acid) could be incorporated into CGP in C. glutamicum.


1985 ◽  
Vol 49 (7) ◽  
pp. 2181-2183 ◽  
Author(s):  
Shin''ichi KUROSAWA ◽  
Kazuharu KATOH ◽  
Hideki YOKOMICHI

1944 ◽  
Vol 79 (1) ◽  
pp. 9-22 ◽  
Author(s):  
Frank L. Engel ◽  
Helen C. Harrison ◽  
C. N. H. Long

1. In a series of rats subjected to hemorrhage and shock a high negative correlation was found between the portal and peripheral venous oxygen saturations and the arterial blood pressure on the one hand, and the blood amino nitrogen levels on the other, and a high positive correlation between the portal and the peripheral oxygen saturations and between each of these and the blood pressure. 2. In five cats subjected to hemorrhage and shock the rise in plasma amino nitrogen and the fall in peripheral and portal venous oxygen saturations were confirmed. Further it was shown that the hepatic vein oxygen saturation falls early in shock while the arterial oxygen saturation showed no alteration except terminally, when it may fall also. 3. Ligation of the hepatic artery in rats did not affect the liver's ability to deaminate amino acids. Hemorrhage in a series of hepatic artery ligated rats did not produce any greater rise in the blood amino nitrogen than a similar hemorrhage in normal rats. The hepatic artery probably cannot compensate to any degree for the decrease in portal blood flow in shock. 4. An operation was devised whereby the viscera and portal circulation of the rat were eliminated and the liver maintained only on its arterial circulation. The ability of such a liver to metabolize amino acids was found to be less than either the normal or the hepatic artery ligated liver and to have very little reserve. 5. On complete occlusion of the circulation to the rat liver this organ was found to resist anoxia up to 45 minutes. With further anoxia irreversible damage to this organ's ability to handle amino acids occurred. 6. It is concluded that the blood amino nitrogen rise during shock results from an increased breakdown of protein in the peripheral tissues, the products of which accumulate either because they do not circulate through the liver at a sufficiently rapid rate or because with continued anoxia intrinsic damage may occur to the hepatic parenchyma so that it cannot dispose of amino acids.


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