Influence of temperature and growth phase on desaturase activity of the mesophilic yeast Candida lipolytica

1983 ◽  
Vol 61 (4) ◽  
pp. 171-177 ◽  
Author(s):  
G. Ferrante ◽  
Y. Ohno ◽  
M. Kates
Keyword(s):  
Desaturase Activity ◽  
Specific Activity ◽  
Aeration Rate ◽  
C Cells ◽  
Candida Lipolytica ◽  
Influence Of Temperature ◽  
Degree Of Unsaturation ◽  
Microsomal Membranes ◽  
Specific Activities ◽  
Temperature And Growth

Microsomal membranes prepared from Candida lipolytica cells grown at 10 °C had a higher lipid content and degree of unsaturation than membranes prepared from cells grown at 25 °C. The specific activities of stearoyl-CoA (18:0-CoA), oleoyl-CoA (18:1-CoA), and dioleoyl phosphatidylcholine (18:1-PC) desaturases in microsomes of cells grown at either 25 or 10 °C showed maximum values near midlog phase, coinciding with the respective maximum absolute content of linoleic (18:2) in the microsomal preparations. The 18:1-CoA desaturase activity in 10 °C cells was nearly double that in 25 °C cells, while the 18:0-CoA and 18:1-PC desaturases had considerably lower activities in 10 °C cells. An increase in aeration rate (shaking speed, 70 to 130 rpm) resulted in increased proportions of 18:2 (32 to 47%, respectively) in microsomes of cells grown at 25 °C and in increased 18:1-CoA desaturase specific activity (83 to 140 pmol∙min−1∙mg−1); however, no significant changes occurred in 18∙0-CoA or 18:1-PC desaturase activities.

scholarly journals Arachidonate cannot be released directly from diacyl-sn-glycero-3-phosphocholine in thrombin-stimulated platelets

1989 ◽  
Vol 259 (2) ◽  
pp. 333-339 ◽  
Author(s):  
O Colard ◽  
M Breton ◽  
D Pepin ◽  
F Chevy ◽  
G Bereziat ◽  
...  
Keyword(s):  
Specific Activity ◽  
Head Group ◽  
Polar Head Group ◽  
Lipoxygenase Inhibitor ◽  
Phospholipid Molecular Species ◽  
Degree Of Unsaturation ◽  
The Mean ◽  
Specific Activities ◽  
Derivatives Of ◽  
Rat Platelets

The origin of the arachidonate released from platelets on stimulation with thrombin was investigated by comparing the specific activities of released arachidonate and of arachidonoyl-containing phospholipids using rat platelets prelabelled with arachidonate. Quantification of the released arachidonate was determined in the presence of BW 755 C, a dual cyclo-oxygenase/lipoxygenase inhibitor, which was found not to modify the arachidonate mobilization between the platelet phospholipids. The phospholipid molecular species were analysed by h.p.l.c. of diradylglycerol benzoate derivatives of diacyl, alkylacyl and alkenylacyl classes. The labelled/unlabelled arachidonate ratio varied greatly in the phospholipids depending on whether an ether or acyl bond was present in sn-1 position of the glycerol, on the length and degree of unsaturation of this fatty chain and on the polar head group. Between 15 s and 5 min of stimulation by thrombin, the released arachidonate kept a constant specific activity which was considerably lower than the specific activity of diacyl-GPC. The specific activity of the released arachidonate was intermediate between the specific activities of the 16:0-20:4 and 18:0-20:4 species of diacyl-GPI and diacyl-GPE, and corresponded to the mean specific activity of alkylacyl-GPC. The data indicate that the released arachidonate cannot come directly from diacyl-GPC, and that two phospholipids in particular can act as direct precursors of the released arachidonate. These are (1) the alkylacyl-GPC and (2) the diacyl-GPE whose hydrolysis would induce an arachidonate transfer from diacyl-GPC.

An International Collaborative Study to Investigate Standardisation of Hirudin Potency

1993 ◽  
Vol 69 (05) ◽  
pp. 430-435 ◽  
Author(s):  
Colin Longstaff ◽  
Man-Yu Wong ◽  
Patrick J Gaffney
Keyword(s):  
Specific Activity ◽  
Collaborative Study ◽  
Residual Activity ◽  
Quality Standard ◽  
Upper Limit ◽  
Assay Procedure ◽  
Specific Activities ◽  
International Collaborative ◽  
Range Of Values ◽  
International Collaborative Study

SummaryAn international collaborative study has been carried out to investigate the reproducibility of hirudin assays in 13 laboratories using four recombinant hirudins and one natural, sulphated product. A simple assay procedure was proposed involving the titration of α-thrombin with inhibitor and measurement of residual activity using a chromogenic substrate. A standard α-thrombin preparation was supplied to ensure that this reagent was of uniform quality throughout the study. The method appeared to present no difficulties and laboratories reported similar potencies for the 5 hirudin samples, in line with expected values. This gave 200–222 Thrombin Inhibitory Units/ampoule (TIU/ampoule) of lyophilised hirudin, with geometric coefficient of variation (gcv) values ranging from 10.15–15.97%. This corresponds to specific activities of approximately 14,300–15,900 TIU/mg protein. This is close to the upper limit of previously reported values of specific activity. We conclude that the precision of this determination compared with the wider range of values in the literature (8,000–16,000 thrombin inhibitory units [TIU]/mg) results from the use of good quality standard α-thrombin by all laboratories. This study has important implications for hirudin standardisation.

scholarly journals Gut Digestive Function and Microbiome after Correction of Experimental Dysbiosis in Rats by Indigenous Bifidobacteria

2021 ◽  
Vol 9 (3) ◽  
pp. 522
Author(s):  
Lyudmila V. Gromova ◽  
Elena I. Ermolenko ◽  
Anastasiya L. Sepp ◽  
Yulia V. Dmitrieva ◽  
Anna S. Alekseeva ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Digestive Enzymes ◽  
Specific Activity ◽  
Control Group ◽  
Enteropathogenic Escherichia Coli ◽  
Beneficial Bacteria ◽  
Opportunistic Bacteria ◽  
Dyspeptic Symptoms ◽  
Specific Activities ◽  
Impaired Motor Function

In recent years, great interest has arisen in the use of autoprobiotics (indigenous bacteria isolated from the organism and introduced into the same organism after growing). This study aimed to evaluate the effects of indigenous bifidobacteria on intestinal microbiota and digestive enzymes in a rat model of antibiotic-associated dysbiosis. Our results showed that indigenous bifidobacteria (the Bf group) accelerate the disappearance of dyspeptic symptoms in rats and prevent an increase in chyme mass in the upper intestine compared to the group without autoprobiotics (the C1 group), but significantly increase the mass of chyme in the colon compared to the C1 group and the control group (healthy animals). In the Bf group in the gut microbiota, the content of opportunistic bacteria (Proteus spp., enteropathogenic Escherichia coli) decreased, and the content of some beneficial bacteria (Bifidobacterium spp., Dorea spp., Blautia spp., the genus Ruminococcus, Prevotella, Oscillospira) changed compared to the control group. Unlike the C1 group, in the Bf group there was no decrease in the specific activities of maltase and alkaline phosphatase in the mucosa of the upper intestine, but the specific activity of maltase was decreased in the colon chyme compared to the control and C1 groups. In the Bf group, the specific activity of aminopeptidase N was reduced in the duodenum mucosa and the colon chyme compared to the control group. We concluded that indigenous bifidobacteria can protect the microbiota and intestinal digestive enzymes in the intestine from disorders caused by dysbiosis; however, there may be impaired motor function of the colon.

scholarly journals THE Ah PHENOTYPE. SURVEY OF FORTY-EIGHT RAT STRAINS AND TWENTY INBRED MOUSE STRAINS

Genetics ◽  
1982 ◽  
Vol 100 (1) ◽  
pp. 79-87
Author(s):  
Daniel W Nebert ◽  
Nancy M Jensen ◽  
Hisashi Shinozuka ◽  
Heinz W Kunz ◽  
Thomas J Gill
Keyword(s):  
Specific Activity ◽  
Inbred Mouse ◽  
Inbred Mouse Strain ◽  
Mouse Strains ◽  
Ah Receptor ◽  
Inbred Mouse Strains ◽  
Sprague Dawley ◽  
Rat Strains ◽  
Specific Activities ◽  
Inbred Rat

ABSTRACT Forty-four inbred and four randombred rat strains and 20 inbred mouse strains were examined for their Ah phenotype by determining the induction of liver microsomal aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity (EC 1.14.14.1) by intraperitoneal treatment with either β-naphthoflavone or 3-methylcholanthrene. All 48 rat strains were found to be Ah-responsive. The maximally induced hydroxylase specific activities of the ALB/Pit, MNR/Pit, MR/Pit, SHR/Pit, and Sprague-Dawley strains were of the same order of magnitude as the basal hydroxylase specific activities of the ACI/Pit, F344/Pit, OKA/Pit, and MNR/N strains. Six of the 20 mouse strains were Ah-nonresponsive (i.e. lacking the normal induction response and presumably lacking detectable amounts of the Ah receptor). The basal hydroxylase specific activities of the BDL/N, NFS/N, STAR/N, and ST/JN mouse strains were more than twice as high as the maximally induced hydroxylase specific activity of the CBA/HT strain.——To date, 24 Ah-nonresponsive mouse strains have been identified, out of a total of 68 known to have been characterized. The reasons for not finding a single Ah-nonresponsive inbred rat strain—as compared with about one Ah-nonresponsive inbred mouse strain found for every three examined—remain unknown.

Lipids and Δ6-desaturase activity alterations in rat liver microsomal membranes induced by fumonisin B1

Lipids ◽  
2002 ◽  
Vol 37 (9) ◽  
pp. 869-877 ◽  
Author(s):  
W. C. A. Gelderblom ◽  
W. Moritz ◽  
S. Swanevelder ◽  
C. M. Smuts ◽  
S. Abel
Keyword(s):  
Rat Liver ◽  
Desaturase Activity ◽  
Fumonisin B1 ◽  
Microsomal Membranes ◽  
Δ6 Desaturase

scholarly journals Regulation of lipogenic capacity in lactating rats

1982 ◽  
Vol 208 (3) ◽  
pp. 611-618 ◽  
Author(s):  
M R Grigor ◽  
A Geursen ◽  
M J Sneyd ◽  
S M Warren
Keyword(s):  
Mammary Gland ◽  
Fatty Acid ◽  
Malic Enzyme ◽  
Fatty Acid Synthase ◽  
Specific Activity ◽  
Mammary Glands ◽  
Lipogenic Enzymes ◽  
Pregnancy And Lactation ◽  
Specific Activities

1. The rate of mammary-gland lipogenesis measured in vivo from 3H2O was suppressed after decreasing the milk demand by decreasing the number of pups from ten to two or three, as well as by giving diets containing lipid [Grigor & Warren (1980) Biochem. J. 188, 61-65]. 2. The specific activities of the lipogenic enzymes fatty acid synthase, glucose 6-phosphate dehydrogenase and ‘malic’ enzyme increased between 6- and 10-fold in the mammary gland and between 2- and 3-fold in the livers during the first 10 days of lactation. The increases in specific activity coupled with the doubling of liver mass which occurred during pregnancy and lactation resulted in considerable differences in total liver activities when compared with virgin animals. 3. Although consumption of a diet containing 20% peanut oil suppressed the activities of the three lipogenic enzymes in the livers, only the ‘malic’ enzyme was affected in the mammary glands. 4. In contrast, decreased milk demand did not affect the specific activities of any of the liver enzymes, whereas it resulted in suppression of all three lipogenic enzymes of the mammary glands. There was no effect on either the cytoplasmic malate dehydrogenase or the lactate dehydrogenase of the mammary gland. 5. In all the experiments performed, the activity of the fatty acid synthase correlated with the amount of material precipitated by the rabbit antibody raised against rat fatty acid synthase.

scholarly journals Partial purification of presynaptic plasma membrane by immunoadsorption.

1982 ◽  
Vol 94 (1) ◽  
pp. 88-96 ◽  
Author(s):  
G P Miljanich ◽  
A R Brasier ◽  
R B Kelly
Keyword(s):  
Plasma Membrane ◽  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Specific Activity ◽  
Electric Organ ◽  
Partial Purification ◽  
Vesicle Membrane ◽  
Specific Activities ◽  
Active Zones ◽  
Presynaptic Plasma Membrane

During transmitter release, synaptic vesicle membrane is specifically inserted into the nerve terminal plasma membrane only at specialized sites or "active zones." In an attempt to obtain a membrane fraction enriched in active zones, we have utilized the electric organ of the marine ray. From this organ, a fraction enriched in nerve terminals (synaptosomes) was prepared by conventional means. These synaptosomes were bound to microscopic beads by an antiserum to purified electric organ synaptic vesicles (anti-SV). The success of this immunoadsorption procedure was demonstrated by increased specific activities of bead-bound nerve terminal cytoplasmic markers and decreased specific activities of markers for contaminating membranes. To obtain a presynaptic plasma membrane (PSPM) fraction, we lysed the bead-bound synaptosomes by hypoosmotic shock and sonication, resulting in complete release of cytoplasmic markers. When the synaptosomal fraction was surface-labeled with iodine before immunoadsorption, 10% of this label remained bead-bound after lysis, compared with 2% of the total protein, indicating an approximately fivefold enrichment of bead-bound plasma membrane. Concomitantly, the specific activity of bead-bound anti-SV increased approximately 30-fold, indicating an enrichment of plasma membrane which contained inserted synaptic vesicle components. This PSPM preparation is not simply synaptic vesicle membrane since two-dimensional electrophoresis revealed that the polypeptides of the surface-iodinated PSPM preparation include both vesicle and numerous nonvesicle components. Secondly, antiserum to the PSPM fraction is markedly different from anti-SV and binds to external, nonvesicle, nerve terminal components.

THE PHYSIOLOGY OF HOST-PARASITE RELATIONS: IX. FURTHER OBSERVATIONS ON THE ACCUMULATION OF RADIOACTIVE SUBSTANCES AT RUST INFECTIONS

1961 ◽  
Vol 39 (6) ◽  
pp. 1393-1407 ◽  
Author(s):  
Michael Shaw
Keyword(s):  
Specific Activity ◽  
Dry Weight ◽  
Insoluble Fraction ◽  
Pentose Phosphate ◽  
Accumulation Ratio ◽  
X Ray ◽  
Specific Activities ◽  
Leaf Segments ◽  
Wheat Leaves ◽  
Host Parasite

Wang (Can. J. Botany, 38, 635–642 (1960)) concluded that the accumulation of radioactivity observed on radioautographs at infection sites on rusted leaves fed with C14-labelled substances was 'apparent' rather than real. The ‘accumulation ratio’ is defined as the ratio of the specific activities (c.p.m./mg dry weight of intact tissue) of rust-infected to uninfected areas of infected leaves. Theoretical considerations relating to the radioautography of leaves labelled with C14 and to the measurement of ‘accumulation ratios’ by extraction of C14-labelled substances from rusted and uninfected segments of infected leaves, as well as experimental data, show that Wang's conclusion is not generally applicable.Experimentally, it was shown using polymethacrylate C14 sources that differences in distance between sources and X-ray film of the order of 100 μ had no effect on the intensity of autoradiographs. Rust-infected leaves, fed with radioactive glucose, were radiographed between X-ray plates. Localization of radioactivity at infection sites was observed on both ‘dorsal’ and ‘ventral’ radiographs, indicating a real accumulation per unit area. Ventral were more radioactive than dorsal surfaces. The main development of the fungus occurred on the former. Radioautography revealed that C14 from glucose-1-C14, glucose-6-C14, and uniformly labelled glucose fed to excised wheat leaves became localized at 10-day-old rust infections in 2 hours. ‘Accumulation ratios’ calculated from the specific activity of leaf segments remained close to 1.0 for at least 6 hours after introduction of the tracer, but increased to more than 2 after 24 hours. When ‘accumulation ratios’ were calculated from the specific activities of individual pustules (excised with a punch 1 mm in diameter) and interpustular disks, values greater than 1 were observed in 2 hours, thus confirming the results of autoradiography. Differences between the ‘accumulation ratios’ observed with glucose-6-C14 and glucose-1-C14 were consistent with an increased role of the pentose phosphate pathway at infection sites. Incorporation of C14 from uniformly labelled glucose into the alcohol-insoluble fraction of rusted leaf segments was 2.5-fold that in uninfected segments in 6 hours and 3.65-fold in 24 hours. The humin formed during hydrochloric acid hydrolysis accounted for approximately 50% of the activity of the alcohol-insoluble material. The ‘accumulation ratio’ for the alcohol-soluble material was only 1.56 after 24 hours.All the results support the view (Shaw and Samborski, Can. J. Botany, 34, 389–405 (1956)) that there is a quantitative, metabolically dependent accumulation of C14 from radioactive glucose at vigorous rust infections. The relative roles of fungus and host in this process are discussed briefly.

scholarly journals Identification and Function of the pdxY Gene, Which Encodes a Novel Pyridoxal Kinase Involved in the Salvage Pathway of Pyridoxal 5′-Phosphate Biosynthesis in Escherichia coli K-12

1998 ◽  
Vol 180 (7) ◽  
pp. 1814-1821 ◽  
Author(s):  
Yong Yang ◽  
Ho-Ching Tiffany Tsui ◽  
Tsz-Kwong Man ◽  
Malcolm E. Winkler
Keyword(s):  
Escherichia Coli ◽  
De Novo ◽  
Specific Activity ◽  
Salvage Pathway ◽  
Pyridoxal Kinase ◽  
Triple Mutant ◽  
Reading Frame ◽  
E Coli ◽  
Specific Activities ◽  
K 12

ABSTRACT pdxK encodes a pyridoxine (PN)/pyridoxal (PL)/pyridoxamine (PM) kinase thought to function in the salvage pathway of pyridoxal 5′-phosphate (PLP) coenzyme biosynthesis. The observation that pdxK null mutants still contain PL kinase activity led to the hypothesis that Escherichia coli K-12 contains at least one other B6-vitamer kinase. Here we support this hypothesis by identifying the pdxY gene (formally, open reading frame f287b) at 36.92 min, which encodes a novel PL kinase. PdxY was first identified by its homology to PdxK in searches of the complete E. coli genome. Minimal clones of pdxY + overexpressed PL kinase specific activity about 10-fold. We inserted an omega cassette intopdxY and crossed the resultingpdxY::ΩKanr mutation into the bacterial chromosome of a pdxB mutant, in which de novo PLP biosynthesis is blocked. We then determined the growth characteristics and PL and PN kinase specific activities in extracts ofpdxK and pdxY single and double mutants. Significantly, the requirement of the pdxB pdxK pdxY triple mutant for PLP was not satisfied by PL and PN, and the triple mutant had negligible PL and PN kinase specific activities. Our combined results suggest that the PL kinase PdxY and the PN/PL/PM kinase PdxK are the only physiologically important B6vitamer kinases in E. coli and that their function is confined to the PLP salvage pathway. Last, we show thatpdxY is located downstream from pdxH (encoding PNP/PMP oxidase) and essential tyrS (encoding aminoacyl-tRNATyr synthetase) in a multifunctional operon.pdxY is completely cotranscribed with tyrS, but about 92% of tyrS transcripts terminate at a putative Rho-factor-dependent attenuator located in thetyrS-pdxY intercistronic region.

scholarly journals Assessment of radiological risk in the area around surface disposal at Al -Tuwaitha site

2019 ◽  
Vol 14 (30) ◽  
pp. 90-97
Author(s):  
Asia H. Al-Mashhadani
Keyword(s):  
Health Risk ◽  
Hpge Detector ◽  
Specific Activity ◽  
Gamma Dose ◽  
Dose Equivalent ◽  
Radiological Risk ◽  
Dose Rates ◽  
Gamma Dose Rates ◽  
Specific Activities

The gamma dose rates and specific activity of 137Cs, 60Co and 40K insamples of soil taken from places near the landfill radiation at Al-Tuwaitha site were measured using a portable NaI(Tl) detector. Theresults of gamma dose rates in samples were ranged from 52.6nGy.h-1 to 131nGy.h-1. Then the specific activity of 137Cs, 60Co and40K in soil were determined using high pure germanium (HPGe)detector. The specific activities were varied from 1.9 to 115500 Bq.kg-1 for 137Cs, from 6.37 to 616.5 Bq. kg-1 for 60Co, and from 3 to839.5 Bq. kg-1 for 40K. The corresponding health risk for the annualeffective dose equivalent varied from 1.85×10-14 to 15.7mSv/y. Theresults were compared with various international recommendations.

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