Oospora lactic lipase: Isolation and properties

1990 ◽  
Vol 55 (8) ◽  
pp. 2110-2117 ◽  
Author(s):  
Kakhramon Davranov ◽  
Mina J. Tabak ◽  
Dzhamil Ch. Diyarov ◽  
Abdumurod Sattarov ◽  
Kamola A. Gulyamova

Lipase (EC 3.1.1.3) of Oospora lactic UzLM-2 was purified by ammonium sulfate (0.40-0.80) fractionation, gel-filtration on Sephadex G-100, column chromatography on DEAE-Sephadex and CM-Sephadex, gel-filtration on Sephadex G-75 and was finally crystallized in concentrated aqueous solution. Crystalline preparation of the enzyme is homogeneous at disc-electrophoresis and ultracentrifugation. Sedimentation coefficient (S20w) is 3.6 S, isoelectric point (pI) at pH 4.2 and molecular mass of the enzyme is 40-43 kD. Amino acid composition analysis showed that none of sulfur-containing amino acids were detected in the crystalline enzyme, but it contains about 8% of carbohydrates and a small amount of lipids. Lipase from Oospora lactic was most active at pH 7.5 and 37 °C in olive oil, stable in the range of pH from 5.7 to 8.0 at 30-40 °C for 18 h and retained stability at 50 °C for 10 min.

1984 ◽  
Vol 222 (3) ◽  
pp. 701-709 ◽  
Author(s):  
R L Olsen ◽  
C Little

The subunit composition of human myeloperoxidase was studied with the use of sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and gel filtration. The subunit pattern observed depended on the manner in which the enzyme was treated before analysis. Reduction before heat treatment in detergent led to two main protein species (Mr 57 000 and 10 500), whereas reduction during or after heat treatment yielded an additional species of Mr 39 000. Heating without any reductive pretreatment yielded the 39 000-Mr form as the major electrophoretic species. Carbohydrate staining showed large amounts of sugar on the 57 000-Mr species and little on the 10 500-Mr form. Significant amounts of haem were associated with this latter subunit. Haem also seemed to be associated with the 57 000-Mr form but not with the 39 000-Mr one. These three subunit forms were isolated and their amino acid composition analysed. The 57 000-Mr and 39 000-Mr forms had very similar amino acid composition and yielded an apparently identical collection of fragments on incubation with CNBr. Once separated, the subunits could not be interconverted. Generally, minor amounts of other molecular-mass forms were observed. The nature of the various molecular-mass forms originating from myeloperoxidase is discussed.


1969 ◽  
Vol 47 (5) ◽  
pp. 679-685 ◽  
Author(s):  
A. J. Finlayson ◽  
R. S. Bhatty ◽  
C. M. Christ

Two proteins, previously described by the authors as BI (S020, buffer 12 S) and AIVS (S020, w 1.7 S) have been isolated from eight varieties of rapeseed belonging to Brassica campestris L. and Brassica napus L. species. These proteins have similar chromatographic and electrophoretic characteristics but differ in amino acid composition, particularly with regard to the sulfur-containing amino acids. One of the rapeseed samples was obtained from plants grown on sulfur-deficient soil. The sulfur deficiency produced, in the mature seed, a much reduced protein content and appeared to affect the structure of the protein BI.


2021 ◽  
Vol 4 (1) ◽  
pp. 56-72
Author(s):  
Mykhailo Peresichnyi ◽  
Svitlana Peresichna ◽  
Anna Sobko

The topicality. To date, the structure of the population’s diet has significant deviations from the formula of a balanced diet in terms of protein intake, including sulfur-containing, which causes the formation of risk factors for the development alimentary and alimentary-dependent diseases. For the prevention of diseases caused by protein deficiency, it is promising to increase its content in vegetable dishes due to the integrated use of raw materials and dietary supplements of high biological value. It is advisable to use food combinatorics in the innovative technologies development for vegetable dishes of improved amino acid composition. The purpose of the study is food combinatorics of amino acid composition of potato croquettes with soy flour, wheat germ; natural shrimp powder “Rieber Food Ingredients” and lentil-spirulina filling in terms of content and balance of essential and substitute amino acids and the degree of their assimilation by the human body. Research methods. Physicochemical, mathematical and statistical methods of experimental data processing with the use of information technologies, methods of ion exchange liquid column chromatography and qualimetric methods have been used. Results. Research is aimed at the use of food combinatorics and scientific substantiation of the amino acid composition of potato croquettes with the use of protein vegetable and non-fish water raw materials and dietary supplements. As a result of scientific researches the food combinatorics at development of technology of vegetable dishes has been carried out, the amino acid structure has been proved and experimentally generalized, the amino acid score and balance of sulfur-containing amino acids in the complex use of potato mass with wheat germ, soy flour, natural shrimp powder “Rieber Food Ingredients” and lentil-spirulina filling in culinary products have been analyzed. The social effect of fuller use of vegetable, non-fish water raw materials, expansion of the range of vegetable dishes with improved amino acid composition and consumer properties of potato dishes in restaurants, preservation and protection of public health has been confirmed. Conclusions and discussions. Food combinatorics was carried out during the potato croquet technology development with the use of protein vegetable and non-fish water raw materials, which contributed to the improvement of the amino acid composition of vegetable dishes and, in particular, to the increase of their biological value.


1981 ◽  
Vol 27 (10) ◽  
pp. 1712-1716 ◽  
Author(s):  
J Menyhárt ◽  
J Gróf

Abstract The aim of the present investigation was to collect information on the molecular composition of uremic "middle molecules," the 500-5000 molecular-mass serum constituents assumed to be involved in the molecular etiology of uremic intoxication. For this purpose, three fractions of serum containing such molecules were separated by cation-exchange column chromatography. These fractions absorb at 240 nm and are characteristically increased in chronic uremia. By one-dimensional paper chromatography of these fractions it was demonstrated that each could be resolved into at least seven (altogether, 21) ninhydrin-positive subfractions. These, like the original fractions, yielded amino acids on acid hydrolysis. The qualitative amino acid composition of the subfractions differed substantially, both from each other and from that of 31 known peptides with which they were compared. We conclude that hitherto unknown peptides with only limited diffusibility through hemodialysis membranes constitute the main bulk of uremic middle molecules.


1986 ◽  
Vol 164 (6) ◽  
pp. 2038-2048 ◽  
Author(s):  
W Strittmatter ◽  
P J Hitchcock

We have presented a method for the extraction and isolation of the gonococcal H.8 antigen. There was no evidence of contamination by other gonococcal proteins, phospholipids, or LPS. The purified H.8 antigen was subjected to preliminary analysis and appeared to be a proteolipid consisting of both protein and lipid components. The amino acid composition was unusual; the peptide portion of the antigen was an alanine and proline-rich molecule that lacked aromatic and sulfur-containing amino acids. The overall amino acid composition is hydrophobic. A lipid constituent was also identified; it was made up of at least two lipid components, which were unique to the H.8 molecule. The chemical nature of the association of the protein and lipid is presently unknown, but it is clearly a tenacious one.


1970 ◽  
Vol 48 (10) ◽  
pp. 1096-1103 ◽  
Author(s):  
L. A. Goding ◽  
R. S. Bhatty ◽  
A. J. Finlayson

A 12 S globulin was isolated from each of the two species of rapeseed (B. napus L. and B. campestris L.) and they have been shown to be similar in terms of amino acid composition, amino terminal amino acids, number of subfractions, and carbohydrate content. One of the major proteins, a glycoprotein, present in each of the 12 S aggregates, was isolated and purified. Its amino acid composition, carbohydrate content, N-terminal amino acids, and sedimentation coefficient are reported herein.


1977 ◽  
Vol 161 (2) ◽  
pp. 389-398 ◽  
Author(s):  
P F Fox ◽  
J R Whitaker

Sheep pepsin was isolated (approx. 120-fold purification) from aqueous abomasal homogenates by (1) pH fractionation, (2) chromatography on Sepharose 4B-poly-L-lysine columns and (3) gel filtration on Sephadex G-100. The enzyme had mol.wt. approx. 34000, N-terminal valine and C-terminal alanine. The amino acid composition of sheep pepsin was generally similar to that of pig and ox pepsins, with a very low content of basic residues and a high content of acidic and hydroxy-amino acids. The pH optimum for NN-dimethyl-casein and NN-dimethyl-haemoglobin as substrates was approx. 1.8. The Km and kcat. for NN-dimethyl-haemoglobin were 46micronM and 1100min-1 respectively, and for NN-dimethyl-casein the corresponding parameters were 50micronM and 420min-1. These values were generally similar to those for pig and ox pepsins. At the pH optimum of 4.6, the sheep pepsin was about 50% as active on benzyloxycarbonyl-L-histidyl-L-phenyl-alanyl-L-tryptophan ethyl ester as was pig pepsin. The pH optimum for the hydrolysis of N-acetyl-L-phenylalanyl-L-di-iodotyrosine by sheep, ox and pig pepsins was approx. 1.85.


1955 ◽  
Vol 8 (4) ◽  
pp. 537 ◽  
Author(s):  
DH Simmonds

The amino acid composition of 16-hr 6N HCI hydrolysates of three qualities of commercially classified wools has now been determined using the technique of Moore and Stein (1951). In this paper the results obtained on samples of Merino 70's and Corriedale 56's wool are compared with those previously reported for Merino wool of 64's quality. The overall pattern of the amino acid composition of the three wools is similar although small variations between the wools are observed with some of the amino acids.


1984 ◽  
Vol 62 (5) ◽  
pp. 276-279 ◽  
Author(s):  
C. H. Lin ◽  
W. Chung ◽  
K. P. Strickland ◽  
A. J. Hudson

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.


1973 ◽  
Vol 134 (2) ◽  
pp. 431-436 ◽  
Author(s):  
W. Ferdinand ◽  
W. Bartley ◽  
V. Broomhead

Amino acid analyses of mitochondrial membranes are compared with the amino acid composition of whole mitochondria (Alberti, 1964) and found to be very similar except in the cystine content. The composition of the endogenous amino acids found in freshly prepared mitochondria has been established and shown to differ considerably from the amino acid composition of membranes or whole mitochondria. The amino acids produced during anaerobic incubation of mitochondria at pH7.4, on the other hand, resemble the membrane in composition, supporting the view that neutral proteinase activity is responsible for their appearance. Aerobic incubation produces a similar pattern of amino acids except that amino acids such as proline, serine, asparagine, glutamic acid and glutamine, which can be metabolically utilized under aerobic conditions, are present to a smaller extent. The presence of large relative concentrations of endogenous taurine, cysteic acid and oxidized glutathione and the accumulation of taurine during incubation is found. The selective retention of taurine and cysteic acid within the mitochondria is established. It is proposed that the first step in the degeneration of isolated mitochondria results from lipid hydroperoxide accumulation caused by the lack of glutathione reductase in isolated mitochondria.


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