scholarly journals Purification and Substrate Specificities of Two α-l-Arabinofuranosidases from Aspergillus awamori IFO 4033

1998 ◽  
Vol 64 (10) ◽  
pp. 4021-4027 ◽  
Author(s):  
Satoshi Kaneko ◽  
Mitsue Arimoto ◽  
Misako Ohba ◽  
Hideyuki Kobayashi ◽  
Tadashi Ishii ◽  
...  
Keyword(s):  
Culture Filtrate ◽  
The Other ◽  
Hydrolysis Reaction ◽  
Aspergillus Awamori ◽  
Side Chain ◽  
Optimum Temperature ◽  
Substrate Specificities ◽  
Molecular Weights ◽  
Ph Values ◽  
Optimum Ph

ABSTRACT α-l-Arabinofuranosidases I and II were purified from the culture filtrate of Aspergillus awamori IFO 4033 and had molecular weights of 81,000 and 62,000 and pIs of 3.3 and 3.6, respectively. Both enzymes had an optimum pH of 4.0 and an optimum temperature of 60°C and exhibited stability at pH values from 3 to 7 and at temperatures up to 60°C. The enzymes released arabinose from p-nitrophenyl-α-l-arabinofuranoside,O-α-l-arabinofuranosyl-(1→3)-O-β-d-xylopyranosyl-(1→4)-d-xylopyranose, and arabinose-containing polysaccharides but not fromO-β-d-xylopyranosyl-(1→2)-O-α-l-arabinofuranosyl-(1→3)-O-β-d-xylopyranosyl-(1→4)-O-β-d-xylopyranosyl-(1→4)-d-xylopyranose. α-l-Arabinofuranosidase I also released arabinose fromO-β-d-xylopy-ranosyl-(1→4)-[O-α-l-arabinofuranosyl-(1→3)]-O-β-d-xylopyranosyl-(1→4)-d-xylopyranose. However, α-l-arabinofuranosidase II did not readily catalyze this hydrolysis reaction. α-l-Arabinofuranosidase I hydrolyzed all linkages that can occur between two α-l-arabinofuranosyl residues in the following order: (1→5) linkage > (1→3) linkage > (1→2) linkage. α-l-Arabinofuranosidase II hydrolyzed the linkages in the following order: (1→5) linkage > (1→2) linkage > (1→3) linkage. α-l-Arabinofuranosidase I preferentially hydrolyzed the (1→5) linkage of branched arabinotrisaccharide. On the other hand, α-l-arabinofuranosidase II preferentially hydrolyzed the (1→3) linkage in the same substrate. α-l-Arabinofuranosidase I released arabinose from the nonreducing terminus of arabinan, whereas α-l-arabinofuranosidase II preferentially hydrolyzed the arabinosyl side chain linkage of arabinan.

scholarly journals Hydrolysis of penicillins and related compounds by the cell-bound penicillin acylase of Escherichia coli

1969 ◽  
Vol 115 (4) ◽  
pp. 733-739 ◽  
Author(s):  
M. Cole
Keyword(s):  
Escherichia Coli ◽  
Penicillin Acylase ◽  
Side Chain ◽  
Bacterial Cells ◽  
Optimum Temperature ◽  
Phenoxymethyl Penicillin ◽  
Optimum Ph ◽  
Penicillanic Acid ◽  
Hydrolysis Of ◽  
Related Compounds

1. A method is given for the preparation of penicillin acylase by using Escherichia coli N.C.I.B. 8743 and a strain selected for higher yield. The enzyme is associated with the bacterial cells and removes the side chains of penicillins to give 6-amino-penicillanic acid and a carboxylic acid. 2. The rates of penicillin deacylation indicated that p-hydroxybenzylpenicillin was the best substrate, followed in diminishing order by benzyl-, dl-α-hydroxybenzyl-, 2-furylmethyl-, 2-thienylmethyl-, d-α-aminobenzyl-, n-propoxymethyl- and isobutoxymethyl-penicillin. Phenylpenicillin and dl-α-carboxybenzylpenicillin were not substrates and phenoxymethyl-penicillin was very poor. 3. Amides and esters of the above penicillins were also substrates for the deacylation reaction, as were cephalosporins with a thienylmethyl side chain. 4. For the deacylation of 2-furylmethylpenicillin at 21° the optimum pH was 8·2. The optimum temperature was 60° at pH7. 5. By using selection A of N.C.I.B. 8743 and determining reaction velocities by assaying yields of 6-amino-penicillanic acid in a 10min. reaction at 50° and pH8·2, the Km for benzylpenicillin was found to be about 30mm and the Km for 2-furylmethylpenicillin, about 10mm. The Vmax. values were 0·6 and 0·24μmole/min./mg. of bacterial cells respectively.

scholarly journals The Properties of Immobilized Invertase Onto a New Support Material; Poly(Methacrylamide/Maleic Acid) Copolymeric Hydrogel

2018 ◽  
Vol 26 (2) ◽  
pp. 307-328 ◽  
Author(s):  
Hesna Nursevin Öztop ◽  
Fatma Banu Çatmaz ◽  
Dursun Saraydin
Keyword(s):  
Maleic Acid ◽  
Storage Stability ◽  
Maximum Velocity ◽  
Optimum Temperature ◽  
Michaelis Constant ◽  
Ph Values ◽  
Optimum Ph ◽  
And Storage ◽  
And Diffusion

Abstract Poly (methacrylamide / maleic acid) PM/MA and poly (methacrylamide) PM hydrogels were prepared aiming to be used as a support for invertase. Spectrophotometric, thermal analysis methods, swelling and diffusion experiments were used for the characterization of hydrogels. The swelling of PM/MA was higher than that of PM in water. The diffusion of water within the hydrogel was found to be non-Fickian. Invertase was immobilized onto PM and PM/MA (samples named PM-I and PM/MA-I respectively). The optimum pH values were found to be; 6.0, 5.0 and 5.5 for free invertase, PM-I and PM/MA-I respectively. The optimum temperature values were found to be 30 °C, 35 °C and 40 °C for free invertase, PM-I and PM/MA-I respectively. The Michaelis constant (Km) and maximum velocity of the enzymes (Vmax) were Km: 11,75 mM, Vmax: 1,95 μmol min−1 for free invertase, Km: 67,24 mM, Vmax: 60,6 μmol min−1 for PM-I and Km: 74,55 mM, Vmax: 18,12 μmol min−1 for PM/MA-I. PM/MA-I showed excellent thermal, operational and storage stability.

scholarly journals Biochemical Differences Between Alcohol Debydrogenases of Drosophila Melanogaster

1976 ◽  
Vol 29 (4) ◽  
pp. 365 ◽  
Author(s):  
JG Oakeshott
Keyword(s):  
Drosophila Melanogaster ◽  
The Other ◽  
Developmental Changes ◽  
Substrate Specificities ◽  
Ph Values ◽  
Ph Profiles

This paper describes substrate specificities, developmental changes in activity, pH profiles, and heat stabilities of isozymes produced by four Adh genotypes in D. melanogaster. No differences are found in the substrate specificities of isozymes from the different genotypes but studies of the other three properties reveal significant differences between the isozymes. Thus relatively low activities are found among extracts of AdhF Adhn2 larvae and among extracts of AdhF AdhF adults aged 44 days. Also AdhF Adhs and Adhs Adhs extracts have relatively high activities at pH 6� 5, and AdhF Adhn2 extracts have relatively low activities at pH values above 10�0. Finally, extracts of AdhF AdhF and AdhF Adhs are more stable at 40�C than are those of Adhs Adhs and AdhF Adhn2 ?

Extracellular enzymes of Phytophthora infestans: endo-cellulase, β-glucosidases, and 1,3-β-glucanases

1985 ◽  
Vol 31 (1) ◽  
pp. 75-82 ◽  
Author(s):  
Jakob Bodenmann ◽  
Ursula Heiniger ◽  
Hans R. Hohl
Keyword(s):  
Phytophthora Infestans ◽  
Isoelectric Point ◽  
Culture Filtrate ◽  
Extracellular Enzymes ◽  
Biochemical Properties ◽  
The Other ◽  
Molecular Weights ◽  
Glucosidase Ii ◽  
Optimal Ph

An endo-cellulase, two β-glucosidases, and two 1,3-β-glucanases from Phytophthora infestans were partially purified from the culture filtrate and their biochemical properties determined. The molecular weights were estimated by chromatography on Sephacryl S-200 and were 21 000 (endo-cellulase), 160 000 – 230 000 and 32 000 (β-glucosidases I and II), 160 000 – 230 000 and 21 000 (β-glucanases I and II). The optimal pH of the endocellulase was 6.0. The other enzymes showed the following optimal pH and temperature values: β-glucosidase 1,5.5 and 48 °C; β-glucosidase II, 5.25 and 30 °C; 1,3-β-glucanase 1, 7.0and40 °C; and 1,3-β-glucanase II, 4.5 and 45 °C. The β-glucosidase II was unstable above 30 °C, while the other enzymes remained stable to 43 °C. The β-glucosidase I did not show Michaelis–Menten kinetics for p-nitrophenyl-glucopyranoside (pNPG) and gentiobiose as substrates. The extrapolated Km value for pNPG was 1.1 mmol/L and the Km value for cellobiose was 280 mmol/L. The Km values of the β-glucosidase II were 34 mmol/L for pNPG, 340 mmol/L for cellobiose, and 42 mmol/L for gentiobiose. Finally, the Km value of the 1,3-β-glucanase II for laminarin was 0.29 g/L. The isoelectric point of the enzymes were 3.2 (endo-cellulase), 3.3 (β-glucosidase I), 4.7 (β-glucosidase II), and 3.4 (the two 1,3-β-glucanases). At 10 mmol/L, Cu2+ inhibited the β-glucosidase I by 90%, and the β-glucosidase II by about 50%. The 1,3-(3-glucanase II was inhibited 75% by Mn2+ and 35% by Cu2+.

Trans-sialidase Associated with Atherosclerosis: Defining the Identity of a Key Enzyme Involved in the Pathology

2019 ◽  
Vol 20 (9) ◽  
pp. 938-941
Author(s):  
Victor Y. Glanz ◽  
Veronika A. Myasoedova ◽  
Andrey V. Grechko ◽  
Alexander N. Orekhov
Keyword(s):  
Acid Metabolism ◽  
Research Subject ◽  
Disease Pathogenesis ◽  
Sialidase Activity ◽  
Ph Values ◽  
Ldl Particles ◽  
Optimum Ph ◽  
Key Enzyme ◽  
St6gal I

Atherosclerosis is associated with the increased trans-sialidase activity, which can be detected in the blood plasma of atherosclerosis patients. The likely involvement in the disease pathogenesis made this activity an interesting research subject and the enzyme that may perform such activity was isolated and characterized in terms of substrate specificity and enzymatic properties. It was found that the enzyme has distinct optimum pH values, and its activity was enhanced by the presence of Ca2+ ions. Most importantly, the enzyme was able to cause atherogenic modification of lowdensity lipoprotein (LDL) particles in vitro. However, the identity of the discovered enzyme remained to be defined. Currently, sialyltransferases, mainly ST6Gal I, are regarded as major contributors to sialic acid metabolism in human blood. In this mini-review, we discuss the possibility that atherosclerosis- associated trans-sialidase does, in fact, belong to the sialyltransferases family.

Comparison of the Potency of 8-L-Arginine, 8-D-Arginine and 8-D-Homoarginine Vasopressin Analogs with Substituted Phenylalanine in Position 2

1994 ◽  
Vol 59 (6) ◽  
pp. 1439-1450 ◽  
Author(s):  
Miroslava Žertová ◽  
Jiřina Slaninová ◽  
Zdenko Procházka
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
Basic Amino Acid ◽  
The Other ◽  
Side Chain ◽  
Inhibitory Potency ◽  
Phase Synthesis ◽  
Other Hand ◽  
Order Of Magnitude

An analysis of the uterotonic potencies of all analogs having substituted L- or D-tyrosine or -phenylalanine in position 2 and L-arginine, D-arginine or D-homoarginine in position 8 was made. The series of analogs already published was completed by the solid phase synthesis of ten new analogs having L- or D-Phe, L- or D-Phe(2-Et), L- or D-Phe(2,4,6-triMe) or D-Tyr(Me) in position 2 and either L- or D-arginine in position 8. All newly synthesized analogs were found to be uterotonic inhibitors. Deamination increases both the agonistic and antagonistic potency. In the case of phenylalanine analogs the change of configuration from L to D in position 2 enhances the uterotonic inhibition for more than 1 order of magnitude. The L to D change in position 8 enhances the inhibitory potency negligibly. Prolongation of the side chain of the D-basic amino acid in position 8 seems to decrease slightly the inhibitory potency if there is L-substituted amino acid in position 2. On the other hand there is a tendency to the increase of the inhibitory potency if there is D-substituted amino acid in position 2.

scholarly journals Critical importance of pH and collector type on the flotation of sphalerite and galena from a low-grade lead–zinc ore

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Abdolrahim Foroutan ◽  
Majid Abbas Zadeh Haji Abadi ◽  
Yaser Kianinia ◽  
Mahdi Ghadiri
Keyword(s):  
Iron Ore ◽  
Low Grade ◽  
Zinc Recovery ◽  
Flotation Process ◽  
Ph Values ◽  
Lead And Zinc ◽  
Optimum Ph ◽  
Iron And Zinc ◽  
Lead Zinc ◽  
Zinc Concentrate

AbstractCollector type and pulp pH play an important role in the lead–zinc ore flotation process. In the current study, the effect of pulp pH and the collector type parameters on the galena and sphalerite flotation from a complex lead–zinc–iron ore was investigated. The ethyl xanthate and Aero 3418 collectors were used for lead flotation and Aero 3477 and amyl xanthate for zinc flotation. It was found that maximum lead grade could be achieved by using Aero 3418 as collector at pH 8. Also, iron and zinc recoveries and grades were increased in the lead concentrate at lower pH which caused zinc recovery reduction in the zinc concentrate and decrease the lead grade concentrate. Furthermore, the results showed that the maximum zinc grade and recovery of 42.9% and 76.7% were achieved at pH 6 in the presence of Aero 3477 as collector. For both collectors at pH 5, Zinc recovery was increased around 2–3%; however, the iron recovery was also increased at this pH which reduced the zinc concentrate quality. Finally, pH 8 and pH 6 were selected as optimum pH values for lead and zinc flotation circuits, respectively.

scholarly journals Purification, properties and comparative specificities of the enzyme prolyl-transfer ribonucleic acid synthetase from Phaseolus aureus and Polygonatum multiflorum

1965 ◽  
Vol 97 (1) ◽  
pp. 112-124 ◽  
Author(s):  
PJ Peterson ◽  
L Fowden
Keyword(s):  
Carboxylic Acid ◽  
Enzyme Preparation ◽  
Enzymic Activity ◽  
The Other ◽  
Asparagus Officinalis ◽  
Acid Activation ◽  
Phaseolus Aureus ◽  
Substrate Specificities ◽  
Imino Acids ◽  
Ammonium Sulphate Fractionation

1. A prolyl-s-RNA synthetase (prolyl-transfer RNA synthetase) has been purified about 250-fold from seed of Phaseolus aureus (mung bean), a species not producing azetidine-2-carboxylic acid, and more than 10-fold from rhizome apices of Polygonatum multiflorum, a liliaceous species containing azetidine-2-carboxylic acid. The latter enzyme was unstable during ammonium sulphate fractionation. 2. The enzymes exhibited different substrate specificities towards the analogue. That from Phaseolus, when assayed by the ATP-PP(i) exchange, showed azetidine-2-carboxylic acid activation at about one-third the rate with proline. Both labelled imino acids gave rise to a labelled aminoacyl-s-RNA. The enzyme from Polygonatum, however, activated only proline. 3. The enzyme from Polygonatum also formed a labelled prolyl-s-RNA with Phaseolus s-RNA but at a lower rate than when the Phaseolus enzyme was used. No reaction occurred when the Phaseolus enzyme was coupled with Polygonatum s-RNA, and only a very slight one was observed when both enzyme and s-RNA came from Polygonatum. 4. Protein preparations from seeds of Pisum sativum, another species not producing azetidine-2-carboxylic acid, also activated the analogue in addition to proline, whereas those from rhizome and seeds of Convallaria, the species from which the analogue was originally isolated, failed to activate it. However, a liliaceous species not producing the analogue, Asparagus officinalis, activated it. 5. Of the other proline analogues investigated, only 3,4-dehydro-dl-proline and l-thiazolidine-4-carboxylic acid were active with the enzyme preparation from Phaseolus. 6. pH optima of 7.9 and 8.4 were established for the enzymes from Phaseolus and Polygonatum respectively. 7. The Phaseolus enzyme was specific for ATP and PP(i). Mn(2+) partially replaced the requirement for Mg(2+) as cofactor. Preincubation with p-chloromercuribenzoate at a concentration of 0.5mm or higher produced over 99% inhibition of the Phaseolus enzyme. One-half the enzymic activity was destroyed by preheating for 5min. at 62 degrees in tris-hydrochloric acid buffer, pH7.9. 8. All experimental evidence supports the hypothesis that azetidine-2-carboxylic acid and proline are activated by the same enzyme in Phaseolus preparations, whereas the analogue was inactive in all Polygonatum preparations. The possible nature of this different substrate behaviour is discussed.

scholarly journals OSMOTIC PRESSURE STUDY OF PROTEIN FRACTIONS IN NORMAL AND IN NEPHROTIC SUBJECTS

1939 ◽  
Vol 69 (6) ◽  
pp. 819-831 ◽  
Author(s):  
Jaques Bourdillon
Keyword(s):  
Osmotic Pressure ◽  
Normal Serum ◽  
The Other ◽  
Protein Fractions ◽  
Molecular Weights ◽  
Other Hand ◽  
Normal Albumin

In serum of patients with nephrosis both albumin and globulin showed by osmotic pressure nearly double the molecular weights of normal albumin and globulin. In the urines of such patients, on the other hand, both proteins showed molecular weights lower even than in normal serum. The colloidal osmotic pressures were measured by the author's method at such dilutions that the van't Hoff law relating pressures to molecular concentrations could be directly applied. For the albumin and globulin of normal serum the molecular weights found were 72,000 and 164,000 respectively, in agreement with the weights obtained by other methods.

Immobilization and Characterization of Tannase and its Haze-removing

2009 ◽  
Vol 15 (6) ◽  
pp. 545-552 ◽  
Author(s):  
Erzheng Su ◽  
Tao Xia ◽  
Liping Gao ◽  
Qianying Dai ◽  
Zhengzhu Zhang
Keyword(s):  
Kinetic Parameter ◽  
High Activity ◽  
Green Tea ◽  
Storage Stability ◽  
Residual Activity ◽  
Black Tea ◽  
Optimum Temperature ◽  
Oolong Tea ◽  
Optimum Ph

Tannase was effectively immobilized on alginate by the method of crosslinking-entrapment-crosslinking with a high activity recovery of 76.6%. The properties of immobilized tannase were investigated. Its optimum temperature was determined to be 35 ° C, decreasing 10 °C compared with that of free enzyme, whereas the optimum pH of 5.0 did not change. The thermal and pH stabilities of immobilized tannase increased to some degree. The kinetic parameter, Km, for immobilized tannase was estimated to be 11.6 × 10-4 mol/L. Fe2+ and Mn2+ could activate the activity of immobilized tannase. The immobilized tannase was also applied to treat the tea beverage to investigate its haze-removing effect. The content of non-estern catechins in green tea, black tea and oolong tea increased by 52.17%, 12.94% and 8.83%, respectively. The content of estern catechins in green tea, oolong tea and black tea decreased by 20.0%, 16.68% and 5.04%, respectively. The anti-sediment effect of green tea infusion treated with immobilized tannase was significantly increased. The storage stability and reusability of the immobilized tannase were improved greatly, with 72.5% activity retention after stored for 42 days and 86.9% residual activity after repeatedly used for 30 times.

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