Enzymes of the Conversion of Succinate to Glutamate in Extracts of Rumen Microorganisms

1972 ◽  
Vol 50 (1) ◽  
pp. 1-8 ◽  
Author(s):  
B. Emmanuel ◽  
L. P. Milligan

Cell-free extracts of mixed rumen microorganisms were incubated in vitro in order to determine the existence and nature of metabolic conversions leading from succinate to glutamate. When succinate was incubated as the substrate in the presence of reduced ferredoxin (FDH), a product was formed, which reacted with 2,4-dinitrophenylhydrazine to give the hydrazone of succinic semialdehyde as determined by paper chromatography. When succinic semialdehyde was incubated in the presence of FDH and 14C-bicarbonate, fixation of label was catalyzed by the extracts. The labelled product was isolated as an organic acid and identified as 2-hydroxyglutaric acid by means of paper chromatography. In the presence of ammonia and NAD, 2-hydroxyglutarate was aminated to a ninhydrin-reactive compound that was identified as glutamate by paper chromatography.From the information obtained, a new pathway for the synthesis of glutamate in rumen microbes was proposed. This pathway entails the reduction of succinate to succinic semialdehyde, followed by reductive carboxylation of succinic semialdehyde to yield 2-hydroxyglutarate, which is then aminated to glutamate. The pathway would agree with the labelling pattern in glutamate produced in the presence of 14C-bicarbonate by mixed rumen microorganisms.

Metabolites ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 15
Author(s):  
Zhen-Wei Zhang ◽  
Yan-Lu Wang ◽  
Wei-Kang Wang ◽  
Yong-Yang Chen ◽  
Xue-Meng Si ◽  
...  

A class of aliphatic short chain nitrocompounds have been reported as being capable of CH4 reduction both in vitro and in vivo. However, the laboratory evidence associated with the metabolic fate of nitrocompounds in the rumen has not been well documented. The present study was conducted to compare in vitro degradation and metabolism of nitroethane (NE), 2-nitroethanol (NEOH), and 2-nitro-1-propanol (NPOH) incubated with mixed rumen microorganisms of dairy cows. After 10 mM supplementation of nitrocompounds, a serious of batch cultures were carried out for 120 h under the presence of two substrates differing in the ratio of maize meal to alfalfa hay (HF, 1:4; LF, 4:1). Compared to the control, methane production was reduced by 59% in NPOH and by >97% in both NE and NEOH, and such antimethanogenic effects were more pronounced in the LF than the HF group. Although NE, NEOH, and NPOH addition did not alter total VFA production, the rumen fermentation pattern shifted toward increasing propionate and butyrate and decreasing acetate production. The kinetic disappearance of each nitrocompound was well fitted to the one-compartment model, and the disappearance rate (k, %/h) of NE was 2.6 to 5.2 times greater than those of NEOH and NPOH. Higher intermediates of nitrite occurred in NEOH in comparison with NPOH and NE while ammonia N production was lowest in NEOH. Consequently, a stepwise accumulation of bacterial crude protein (BCP) in response to the nitrocompound addition was observed in both the HF and LF group. In brief, both NE and NEOH in comparison with NPOH presented greater antimethanogenic activity via the shift of rumen fermentation. In addition, the present study provided the first direct evidence that rumen microbes were able to cleave these nitrocompounds into nitrite, and the subsequent metabolism of nitrite into ammonia N may enhance the growth of rumen microbes or promote microbial activities.


2020 ◽  
Vol 49 (6) ◽  
pp. 1139-1147
Author(s):  
E.C.B. Costa ◽  
G.G.L. Araújo ◽  
J.S. Oliveira ◽  
E.M. Santos ◽  
L.T. Henriques ◽  
...  

The aim of this study was to evaluate the effects of various concentrations of three salts (sodium chloride (NaCl), magnesium chloride (MgCl2), and calcium chloride (CaCl2)) on the in vitro rumen fermentation of cellulose, starch, and protein substrates. Six salt concentrations were tested, separately, namely 0, 100, 200, 400, 800, and 1600 mg/dL. The experiment was conducted using the completely randomized design in a 6 × 3 × 3 factorial arrangement with main effects of salt concentration and salt type (six levels of three salts (NaCl, MgCl2, or CaCl2) (0, 100, 200, 400, 800, and 1600 mg/dL) into three substrates [starch, cellulose, and glucose]) with three replicates. Cellulose- and glucose-fermenting bacteria were sensitive to NaCl concentrations greater than 400 mg/dL (17.48 decisiemens per metre (dS/m)) and 800 mg/dL (20.55 dS/m) in the media, respectively. In contrast, starch-fermenting bacteria continued to grow in NaCl concentrations up to 1600 mg/dL (29.09 dS/m). Thus, it was concluded that starch-fermenting microorganisms tolerated higher concentrations of NaCl compared with the other microbial groups. Cellulose-fermenting microorganisms are less tolerant to MgCl2 in relation to the other microbial groups. Starch, cellulose-, and glucose-fermenting bacteria from cattle tolerate CaCl2 concentrations of up to 1600 mg/dL (12.26 dS/m). These results suggest that brackish water may be used for ruminants. However, it is important perform an analysis of that water and then to adjust diets to minimize the effects of types of salt and concentrations of salt on rumen microorganisms. Keywords: brackish water, dissolved salts, rumen microbes, water quality


1960 ◽  
Vol XXXIII (III) ◽  
pp. 444-450 ◽  
Author(s):  
Maria de la Luz Suarez Soto ◽  
Jean Legault Démare

ABSTRACT Serum gonadotrophin (PMS) when added to the incubation medium of rat ovary slices increases the amount of Δ4-3-ketosteroids produced. This enhancement is proportional to the logarithm of dose. The ketosteroids were determined by their ultraviolet absorption; paper chromatography has shown that only androst-4-en-3,17-dione is present.


2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 370-370
Author(s):  
Lauren L Kovanda ◽  
Monika Hejna ◽  
Yanhong Liu

Abstract The aim of this experiment was to examine the anti-inflammatory effects of butyric acid, sodium butyrate, monobutyrin and tributyrin using porcine alveolar macrophages (PAMs). PAMs were isolated from the bronchial lavage of 6 piglets at 6 weeks of age, and then seeded at 106 cells/mL in 24-well plates. After 24 h incubation, cells were treated with different treatments in a randomized complete block design with 10 replicates. The treatments were in a factorial arrangement with 2 doses of lipopolysaccharide (LPS, 0 or 1 μg/mL) and 5 levels of organic acid (0, 0.5, 1, 2, 4 mM for butyric acid and tributyrin and 0, 1, 2, 4, 8 mM for sodium butyrate and monobutyrin). Supernatants were collected after another 24 h incubation and analyzed for tumor necrosis factor alpha (TNF-α). Cell viability was also tested by the MTT assay. Data were analyzed using the MIXED procedure of SAS. No cytotoxic effect was observed in LPS challenge and each organic acid with the percentage of live cells was more than 76% in comparison to the sham control. Sodium butyrate at 2 and 4 mM dose exhibited (P < 0.01) a stimulatory effect on cell proliferation. LPS challenge remarkably stimulated (P < 0.0001) TNF-α secretion from PAMs. In the non-challenge group, butyric acid, monobutyrin, and tributyrin linearly reduced TNF-α production from PAMs, whereas 2 mM sodium butyrate tended to increase (P = 0.056) TNF-α secretion from PAMs. In the LPS challenge group, all tested organic acid dose-dependently reduced (P < 0.001) TNF-α production from LPS-challenged PAMs, with the strongest inhibiting effect observed at the highest dose. Results indicated that butyric acid and its derivatives that were tested in the current experiment all had strong anti-inflammatory activities in vitro.


Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 222
Author(s):  
Abdulaziz A. Al-Askar ◽  
WesamEldin I. A. Saber ◽  
Khalid M. Ghoneem ◽  
Elsayed E. Hafez ◽  
Amira A. Ibrahim

Presently, the bioprocessing of agricultural residues to various bioactive compounds is of great concern, with the potential to be used as plant growth promoters and as a reductive of various diseases. Lycopersiconesculentum, one of the most consumed crops in the human diet, is attacked by Fusarium wilt disease, so the main aim is to biocontrol the pathogen. Several fungal species were isolated from decayed maize stover (MS). Trichodermaasperellum was chosen based on its organic acid productivity and was molecularly identified (GenBank accession number is MW195019). Citric acid (CA) was the major detected organic acid by HPLC. In vitro, CA of T.asperellum at 75% completely repressed the growth of Fusariumoxysporum f. sp. lycopersici (FOL). In vivo, soaking tomato seeds in CA enhanced the seed germination and vigor index. T. asperellum and/or its CA suppressed the wilt disease caused by FOL compared to control. There was a proportional increment of plant growth and yield, as well as improvements in the biochemical parameters (chlorophyll pigments, total phenolic contents and peroxidase, and polyphenol oxidase activities), suggesting targeting both the bioconversion of MS into CA and biological control of FOL.


1982 ◽  
Vol 65 (8) ◽  
pp. 1457-1464 ◽  
Author(s):  
J.D. Erfle ◽  
R.J. Boila ◽  
R.M. Teather ◽  
S. Mahadevan ◽  
F.D. Sauer

1969 ◽  
Vol 49 (2) ◽  
pp. 135-141 ◽  
Author(s):  
L. P. Milligan ◽  
A. R. Robblee ◽  
J. C. Wood ◽  
W. C. Kay ◽  
S. K. Chakrabartty

The preparation of a polymer of urea and furfural containing 23.2% nitrogen is described. This product was converted by rumen microorganisms in vitro to ammonia at a rate approximately one-seventh that of conversion of urea to ammonia. Use of the polymer as a dietary supplement in a feeding trial with lambs improved nitrogen retention over that of unsupplemented controls by 3.45 g of nitrogen retained per day, while an isonitrogenous quantity of supplemental urea improved nitrogen retention by 0.51 g of nitrogen retained per day. The blood urea pattern, throughout the day, of lambs adapted to control, urea-supplemented and urea–furfural polymer-supplemented rations indicated a slow, prolonged production of ammonia from the latter supplement and very rapid, short-term degradation of urea in vivo.


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