SOME ENZYMIC PROPERTIES OF A PARTIALLY PURIFIED AMINOACYLASE OBTAINED FROM A POLISH VARIETY OF RAPESEED (BRASSICA CAMPESTRIS L.)

1961 ◽  
Vol 39 (5) ◽  
pp. 843-853 ◽  
Author(s):  
K. Ozaki ◽  
L. R. Wetter

Aminoacylase, which hydrolyzes N-acetyl amino acids, has been demonstrated in rapeseed. The enzyme was purified 150-fold by fractionation with ammonium sulphate and calcium phosphate gel. The purified preparation hydrolyzed N-acetyl amino acids and in addition certain dipeptides and chloracetyl-L-tyrosine. The enzyme was stable at −20 °C and had a wide pH optimum (7.2 to 8.8). Cobalt ion was found to be an activator, while sulphydryl-reacting agents such as p-chloromercuribenzoate and some metal-chelating agents inhibited the hydrolysis. The enzyme showed rigorous specificity for the L-isomer. A comparison of the ratio of activities obtained for different enzyme preparations indicates that more than one enzyme is concerned in the hydrolysis of the different substrates.

1960 ◽  
Vol 38 (2) ◽  
pp. 125-134 ◽  
Author(s):  
K. Ozaki ◽  
L. R. Wetter

Glyceric acid kinase from Polish variety rapeseed (Brassica campestris L.) was purified approximately 30-fold by ammonium sulphate fractionation and adsorption on alumina Cγ gel. The enzyme has a pH optimum at 7.9 and a pH stability range extending from 6.5 to 7.5. The maximum temperature for the reaction was 45 °C. The phosphorylation required ATP and a metal ion; Mg++ was slightly more effective than Mn++ and Co++, while Ni++ and Zn++ were ineffective. The Michaelis constants for the Mg–ATP complex and the substrate were 7.3 × 10−4 M and 1.6 × 10−3 M respectively. The reaction products, ADP and 3-phosphoglyceric acid, inhibited the phosphorylation. Sulphydryl reagents such as p-chloromercuribenzoate, o-iodosobenzoate, N-ethylmaleimide, and iodo-acetate completely inhibited the enzyme at low concentrations. 3-Phosphoglyceric acid was isolated and characterized from the enzyme reaction mixture.


Author(s):  
С.А. БИРЮКОВА ◽  
Н.М. АГЕЕВА ◽  
М.Г. МАРКОВСКИЙ ◽  
Е.Н. ГОНТАРЕВА

Исследовано влияние ферментных препаратов Лалзайм ИЭКС-ВИ, Колор энзим (оба производства Франции) и Тренолин руж (Германия) на ферментацию виноматериалов из винограда сорта Каберне-Совиньон. Установлено, что внесение ферментных препаратов Лалзайм ИЭКС-ВИ и Колор энзим в дозировке в 3–4 раза меньшей, чем дозировка препарата Тренолин руж, способствует увеличению выхода сусла и его насыщению компонентами фенольного комплекса, в том числе красящими веществами. При применении препаратов Лалзайм ИЭКС-ВИ и Колор энзим также снижаются дозировки других вспомогательных материалов – бентонита и желатина и потери вина с клеевыми осадками. Образцы виноматериала, обработанные препаратом Лалзайм ИЭКС-ВИ, были наиболее устойчивы к коллоидным помутнениям. Установлено, что применение ферментных препаратов способствует глубокому гидролизу комплексов биополимеров вина, в том числе белково-полифенольно-полисахаридных, до низкомолекулярных соединений – аминокислот, углеводов и фенольных соединений. Установлено, что при батонаже заметно улучшается качество вина (появление мягкости во вкусе, снижение проявления кислотности, формирование яркого сортового аромата), но концентрация фенольных соединений, в том числе красящих веществ, равномерно уменьшается, особенно на 3-м месяце выдержки. Поэтому продолжительность батонажа в технологии красных столовых вин следует ограничить двумя месяцами. Influence of enzyme preparations Lalzim IEX-VI, Kolor enzyme (both from France) and Trenolin Rouge (Germany) to the fermentation of wine materials from grapes of Cabernet-Sauvignon was investigated. Introduction of enzyme preparations Lalzim IEX-VI and Kolor enzyme in a dosage of 3–4 times smaller than the dosage of the preparation Trenolin Rouge, increases the yield of the wort and saturation components of the phenolic complex, including colorants. When using preparations Lalzim IEX-VI and Kolor enzyme dosage other support materials – bentonite and gelatin and the loss of wine from glutinous rainfall are also decrease. The samples of wine processed by drug Lalzim IEX-VI were the most resistant to colloidal opacities. It was found that the use of enzyme preparations promotes deep hydrolysis of complexes of wine biopolymers, including protein-polyphenol-polysaccharide, to low molecular compounds – amino acids, carbohydrates and phenolic compounds. It is established that the quality of the wine when batonnage is distinctly improved (the appearance of softness on the palate, decreasing the appearance of acidity, the formation of bright varietal aroma), but the concentration of phenolic compounds, including pigments, are evenly reduced, particularly at 3d months of exposure. Therefore, the duration batonnage of in the technology of red table wines should be limited to two months.


1960 ◽  
Vol 38 (1) ◽  
pp. 125-134 ◽  
Author(s):  
K. Ozaki ◽  
L. R. Wetter

Glyceric acid kinase from Polish variety rapeseed (Brassica campestris L.) was purified approximately 30-fold by ammonium sulphate fractionation and adsorption on alumina Cγ gel. The enzyme has a pH optimum at 7.9 and a pH stability range extending from 6.5 to 7.5. The maximum temperature for the reaction was 45 °C. The phosphorylation required ATP and a metal ion; Mg++ was slightly more effective than Mn++ and Co++, while Ni++ and Zn++ were ineffective. The Michaelis constants for the Mg–ATP complex and the substrate were 7.3 × 10−4 M and 1.6 × 10−3 M respectively. The reaction products, ADP and 3-phosphoglyceric acid, inhibited the phosphorylation. Sulphydryl reagents such as p-chloromercuribenzoate, o-iodosobenzoate, N-ethylmaleimide, and iodo-acetate completely inhibited the enzyme at low concentrations. 3-Phosphoglyceric acid was isolated and characterized from the enzyme reaction mixture.


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.


MRS Advances ◽  
2020 ◽  
Vol 5 (52-53) ◽  
pp. 2669-2678
Author(s):  
Jeovani González P. ◽  
Ramiro Escudero G

AbstractDeinking of recycled office (MOW) paper was carried out by using a flotation column and adding separately sodium hydroxide, and the enzyme Cellulase Thricodema Sp., as defibrillators.The de-inked cellulose fibers were characterized according to the standards of the paper industry, to compare the efficiency of the deinking of each chemical reagent used to hydrolyze the fibers and defibrillate them.The computational simulation of the molecular coupling between the enzyme and cellulose was performed, to establish the enzyme-cellulose molecular complex and then to identify the principal amino-acids of endo-β-1,4-D-glucanase in this molecular link, which are responsible for the hydrolysis of the cellulose.Experimental results show the feasibility to replace sodium hydroxide with the enzyme Cellulase Thricodema Sp., by obtaining deinked cellulose with similar optical and physical properties.The use of the enzyme instead of sodium hydroxide avoids the contamination of the residual water; in addition to that, the column is operated more easily, taking into consideration that the pH of the system goes from alkaline to neutral.


Author(s):  
Е. M. Serba ◽  
М. B. Overchenko ◽  
L. V. Rimareva ◽  
N. I. Ignatova ◽  
А. E. Orekhova ◽  
...  

In the production of alcohol in the preparation of grain raw materials for fermentation, the main role is given to enzyme preparations of amylolytic action, which are key enzymes that catalyze the hydrolysis of starch. Amylolytic enzyme preparations with a different composition of enzymes and their level of activity, a mechanism of biocatalytic effect on starch, and a range of thermal and pH optimum are widely represented on the Russian market. The development of optimal conditions for the preparation of grain wort, the rational selection and dosage of concentrated enzyme preparations, the properties of which correspond to the parameters of the technological process, will ensure the effective preparation of starch for fermentation, and increase the profitability of alcohol production. The aim of this work was to study the influence of enzyme preparations of amylolytic action and the conditions of their use on the efficiency of the process of alcoholic fermentation and the yield of the final product, ethanol. The effect of various dosages of enzyme preparations of glucoamylase action, with a different ratio of the main enzyme glucoamylase and minor enzyme α-amylase, as well as methods for preparing wheat wort on the process of alcoholic fermentation, was studied. It was found that the enzyme preparation, the source of glucoamylase, in which α-amylase was present in a ratio of 15: 1 (in terms of activity level), turned out to be more effective in fermenting prepared wheat wort: its optimal dosage was 8 units. GLS / g starch. The presence of a sufficient amount of α-amylase in this preparation compensated for the dosage of thermostable α-amylase. The alcohol concentration in the mash was 10.2% vol., The alcohol yield was 67.9 cm3 / 100 g of starch. When glucoamylase with a lower ratio of the main and minor enzyme (75: 1) was used at the saccharification stage, an increase in the wort fermentation depth was observed with an increase in the concentration of glucoamylase to 9-10 units of GLS / g and α-amylase to 0.5 units. AC / g. It was also found that an increase in the duration of enzymatic-hydrolytic preparation of the wort had a positive effect on the fermentation process, the alcohol concentration in the mash increased to 10.2 vol.%. It was shown that the introduction of proteases into the wort helps to reduce the viscosity of grain wort, enriching it with assimilable yeast amino acids, which leads to an increase in the yield of alcohol. It has been confirmed that the synergy of the action of enzymes of amylolytic and proteolytic effects on polymers of grain raw materials allows to increase the efficiency of their conversion to ethanol. The conditions of enzymatic-hydrolytic processing of grain raw materials for fermentation are developed. The use of the digestion stage did not significantly affect the fermentation results of wheat wort.


2018 ◽  
Vol 69 (10) ◽  
pp. 2794-2798
Author(s):  
Alina Diana Panainte ◽  
Ionela Daniela Morariu ◽  
Nela Bibire ◽  
Madalina Vieriu ◽  
Gladiola Tantaru ◽  
...  

A peptidic hydrolysate has been obtained through hydrolysis of bovine hemoglobin using pepsin. The fractioning of the hydrolysate was performed on a column packed with CM-Sepharose Fast Flow. The hydrolysate and each fraction was filtered and then injected into a HPLC system equipped with a Vydak C4 reverse phase column (0.46 x 25 cm), suitable for the chromatographic separation of large peptides with 20 to 30 amino acids. The detection was done using mass spectrometry, and the retention time, size and distribution of the peptides were determined.


1980 ◽  
Vol 45 (4) ◽  
pp. 1099-1108 ◽  
Author(s):  
Mikuláš Chavko ◽  
Michal Bartík ◽  
Evžen Kasafírek

A polarographic study of the hydrolysis of [8-lysine]vasopressin and some hormonogens of the vasopressin series with the blood serum of women in the last week of pregnancy was studied. The dependence of hydrolysis on pH (pH optimum: 7.4-7.50, substrate concentration (Km 1.2 . 10-5M), pH stability and thermal stability were determined. The rate of hydrolysis of individual vasopressin analogues decreases in the order: [8-lysine]vasopressin > Nα-glycyl-prolyl[8-lysine]-vasopressin > Nα-leucyl-[8-lysine]vasopressin > Nα-alanyl-[8-lysine]vasopressin > Nα-phenyl alanyl-[8-lysine]vasopressin > Nα-diglycyl-[8-lysine]vasopressin > Nα-prolyl-[8-lysine]vasopressin > Nα-triglycyl-[8-lysine]vasopressin > Nα-sarcosyl-glycyl-[8-lysine]vasopressin. The degree of hydrolysis gradually increases to a multiple with the length of the pregnancy in consequence of the presence of oxytocine. However, vasopressin is also hydrolysed to a small extent with the enzymes from the blood sera of non-pregnant women. Under similar analytical conditions oxytocin was not hydrolysed with the sera of non-pregnant women and therefore oxytocin is a more suitable substrate than vasopressin for polarographic determination of serum oxytocinase.


1972 ◽  
Vol 247 (18) ◽  
pp. 5746-5752
Author(s):  
Ferenc J. Kézdy ◽  
Satya P. Jindal ◽  
Myron L. Bender

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