scholarly journals Production and optimization of Aspergillus niger glucoamylase using amylopectin from guinea corn starch as the sole carbon source

2017 ◽  
Vol 52 (4) ◽  
pp. 263-272
Author(s):  
PC Okwuenu ◽  
AL Ezugwu ◽  
FC Chilaka
Keyword(s):  
Carbon Source ◽  
Ammonium Sulphate ◽  
Sole Carbon Source ◽  
Corn Starch ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Glucoamylase Activity ◽  
Ammonium Sulphate Precipitation ◽  
Specific Activities ◽  
Two Peaks

A Fourteen day experimental study was carried out to determine the day of highest glucoamylase activity using amylopectin from guinea corn starch as the sole carbon source. Two peaks of high activity were observed on the fifth and twelveth days, and were thus mass produced. Specific activities for crude enzymes were found to be 729.45 U/mg and 1046.82 U/mg for day five and twelve harvested enzymes respectively. Ammonium sulphate saturations, 70% and 20%, were found suitable to precipitate proteins with highest glucoamylase activity for day five and twelve harvested enzymes respectively. After ammonium sulphate precipitation and gel filtration, specific activities were found to be 65.98 U/mg and 180.52 U/mg respectively for day five harvested enzyme and 61.51 U/mg and 272.81 U/mg for day twelve harvested enzyme. The pH optimum for day five harvested enzyme were found to be 7.5, 7.5 and 6.0 using tiger nut, cassava and guinea corn starches as substrates respectively, also, the pH optimum for day twelve harvested enzyme were found to be 5.0, 8.5 and 7.0 using tiger nut, cassava and guinea corn starches as substrate, respectively. Optimum temperatures were found to be 50˚C and 45˚C for day five and twelve harvested enzymes, respectively. Km and Vmax, of day five harvested enzyme were found to be 770.75 mg/ml and 2500 μmol/min, 158.55 mg/ml and 500 μmol/min and 46.23 mg/ml and 454.53 μmol/min using cassava, guinea corn and tiger nut starches as substrate respectively. Km and Vmax of day twelve harvested enzyme were found to be 87.1 mg/ml and 384.61 μmol/min, 29.51 mg/ml and 243.90 μmol/min, and 2364 mg/ml and 2500 μmol/min, using cassava, guinea corn and tiger nut starches as substrate respectively.Bangladesh J. Sci. Ind. Res. 52(4), 263-272, 2017

scholarly journals Canine submandibular-gland hyaluronidase. Purification and properties

1968 ◽  
Vol 110 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Y. H. Tan ◽  
J. M. Bowness
Keyword(s):  
Submandibular Gland ◽  
Ammonium Sulphate ◽  
Rat Liver ◽  
Gel Filtration ◽  
Acid Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Purification And Properties ◽  
Freeze Dried ◽  
Specific Activities

1. Methods for the purification of dog submandibular-gland hyaluronidase from sedimentable and non-sedimentable portions of a homogenate and from the whole homogenate are presented. The method consists of three main steps: removal of mucin by acid precipitation or gel filtration on Sephadex G-200, ammonium sulphate precipitation and CM-cellulose chromatography. By this method specific activities of up to 1·28 and 0·78μmoles of N-acetylglucosamine/min./mg. of protein were obtained for the purified freeze-dried non-sedimentable hyaluronidase and for the sedimentable hyaluronidase respectively. 2. A comparison of some of the properties of the non-sedimentable and the sedimentable hyaluronidase preparation indicated that there was little difference between the two and that they both resembled lysosomal hyaluronidase from rat liver.

Fermentative Production, Purification and Characterization of Nisin

2008 ◽  
Vol 4 (5) ◽  
Author(s):  
Swapnali S Gujarathi ◽  
Sandip B. Bankar ◽  
Laxmi A. Ananthanarayan
Keyword(s):  
Ammonium Sulphate ◽  
Hydrophobic Interaction ◽  
Orthogonal Array ◽  
Gel Filtration ◽  
Statistical Design ◽  
Temperature Stability ◽  
Gel Chromatography ◽  
Purification And Characterization ◽  
Ammonium Sulphate Precipitation

Bacteriocins are bactericidal or bacteriostatic in action and active against closely related species. Nisin is the bacteriocin produced by the lactic acid bacteria (LAB). Nisin is a small (3353 Da), cationic, hydrophobic, and 34-amino acid peptide. It is used in products such as pasteurized processed cheese, salad dressing, and liquid whole eggs to inhibit the growth of Gram-positive microorganisms including Listeria monocytogenes. The objective of the present work was to study production, purification and characterization of nisin. The production of nisin was carried out using Lactococcus lactis subsp lactis MTCC 440 by one-factor-at-a-time method and statistical design (Orthogonal array). Purification was carried out using ammonium sulphate precipitation followed by hydrophobic interaction and gel chromatography. Characterization was done for pH and temperature stability. The activity of nisin after one factor at a time optimization was found to be 5120 AU/ml. The activity of the nisin increased to 6800 AU/ml after optimization by Orthogonal Array design. Ammonium sulphate precipitation gives good yield of nisin with 60 to 80% saturation with 2.52 fold purity. The overall purification by hydrophobic interaction and gel filtration chromatography was 10.87 fold with 50.84% yield and 8.8 fold with 49.65% yield as compared to crude broth respectively.

Meta-Methylation of Flavonol Rings A (8-) and B (3'-) Is Catalysed by Two Distinct O-Methyltransferases in Lotus corniculatus

1983 ◽  
Vol 38 (5-6) ◽  
pp. 413-417 ◽  
Author(s):  
Maurice Jay ◽  
Vincenzo De Luca ◽  
Ragai Ibrahim
Keyword(s):  
Ammonium Sulphate ◽  
Biochemical Markers ◽  
Flower Colour ◽  
Lotus Corniculatus ◽  
The Other ◽  
Flower Buds ◽  
Ph Optimum ◽  
Genetic Studies ◽  
Ammonium Sulphate Precipitation ◽  
Optimum Ph

Two O-methyltransferases specific for flavonol rings A and B were isolated from young flower buds of Lotus corniculatus. They were partially purified by ammonium sulphate precipitation and successive chromatography on Sephadex G-100 and Polybuffer ion exchanger. One enzyme focused at pI 5.5 and catalysed the O-methylation of position 8 of flavonols with a pH optimum of 8.1. The other enzyme had a pi of 5.1 and preferentially attacked position 3' at an optimum pH of 7.7. The methylated products of both enzymes seem to contribute to the flower colour of Lotus and may be used as biochemical markers in genetic studies of this genus.

scholarly journals Biobleaching of Industrial Important Dyes with Peroxidase Partially Purified from Garlic

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Akudo Chigozirim Osuji ◽  
Sabinus Oscar O. Eze ◽  
Emmanuel Emeka Osayi ◽  
Ferdinand Chiemeka Chilaka
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Low Cost ◽  
Gel Filtration ◽  
Enzyme Concentration ◽  
Gel Filtration Chromatography ◽  
Vat Dyes ◽  
Ammonium Sulphate Precipitation ◽  
Ph Range ◽  
Better Than

An acidic peroxidase was extracted from garlic (Allium sativum) and was partially purified threefold by ammonium sulphate precipitation, dialysis, and gel filtration chromatography using sephadex G-200. The specific activity of the enzyme increased from 4.89 U/mg after ammonium sulphate precipitation to 25.26 U/mg after gel filtration chromatography. The optimum temperature and pH of the enzyme were 50°C and 5.0, respectively. The Km andVmaxfor H2O2and o-dianisidine were 0.026 mM and 0.8 U/min, and 25 mM and 0.75 U/min, respectively. Peroxidase from garlic was effective in decolourizing Vat Yellow 2, Vat Orange 11, and Vat Black 27 better than Vat Green 9 dye. For all the parameters monitored, the decolourization was more effective at a pH range, temperature, H2O2concentration, and enzyme concentration of 4.5–5.0, 50°C, 0.6 mM, and 0.20 U/mL, respectively. The observed properties of the enzyme together with its low cost of extraction (from local sources) show the potential of this enzyme for practical application in industrial wastewater treatment especially with hydrogen peroxide. These Vat dyes also exhibited potentials of acting as peroxidase inhibitors at alkaline pH range.

scholarly journals Cathepsin D. Purification of isoenzymes from human and chicken liver

1970 ◽  
Vol 117 (3) ◽  
pp. 601-607 ◽  
Author(s):  
A. J. Barrett
Keyword(s):  
Isoelectric Focusing ◽  
Cathepsin D ◽  
Ph Dependence ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Ph Optimum ◽  
Specific Activities ◽  
Acetone Fractionation ◽  
High Degree

1. The Barrett (1967) assay for cathepsin D was slightly modified. 2. The enzyme was purified from liver of man and chicken by a procedure involving autolysis, acetone fractionation, ion-exchange chromatography and isoelectric focusing. 3. Several isoenzymes of cathepsin D were resolved in the isoelectric-focusing step, and three major forms, α,β and γ, were distinguished for each species. 4. A modified analytical method of isoelectric focusing in polyacrylamide gel indicated a high degree of homogeneity of the purified β and γ isoenzymes from each species, and this was supported by their constant high specific activities. 5. Gel filtration of the isoenzymes in a calibrated column of Sephadex G-100 showed that each had a molecular weight of 45000. 6. Human cathepsin D had a pH optimum of 3.5, and that of chicken enzyme was 3.0, haemoglobin being used as substrate. In each species, the three isoenzymes have the same pH-dependence curve. 7. The purified cathepsin D samples showed very little action on acid-denatured albumin.

scholarly journals Isopentenyl pyrophosphate isomerase from liver

1968 ◽  
Vol 106 (4) ◽  
pp. 835-840 ◽  
Author(s):  
P. W. Holloway ◽  
G. Popják
Keyword(s):  
Ammonium Sulphate ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Pig Liver ◽  
Isopentenyl Pyrophosphate ◽  
Km Value ◽  
Ammonium Sulphate Fractionation ◽  
Dimethylallyl Pyrophosphate

Isopentenyl pyrophosphate isomerase (EC 5.3.3.2) was purified from extracts of pig liver by ammonium sulphate fractionation and by gel filtration. After about 20-fold purification the preparations were free of phosphatase and prenyltransferase (EC 2.5.1.1), the two enzymes that could have interfered with the assays. The isomerase has a distinct pH optimum at 6·0 and is activated by Mn2+ in preference to Mg2+. The Km value for isopentenyl pyrophosphate is 4×10−6m. The equilibrium of the reaction favours the formation of dimethylallyl pyrophosphate. The reversibility of the isomerase reaction was demonstrated directly by the formation of isopentenyl pyrophosphate from dimethylallyl pyrophosphate. It is suggested that two prenyl isomerases might exist, one involved in the synthesis of trans- and another in the synthesis of cis-polyprenyl substances.

scholarly journals Microbial oxidation of amines. Partial purification of a trimethylamine mono-oxygenase from Pseudomonas aminovorans and its role in growth on trimethylamine

1974 ◽  
Vol 140 (2) ◽  
pp. 253-263 ◽  
Author(s):  
Christopher A. Boulton ◽  
M. James C. Crabbe ◽  
Peter J. Large
Keyword(s):  
Carbon Source ◽  
Oxidation Product ◽  
Sole Carbon Source ◽  
Secondary Amines ◽  
Partial Purification ◽  
Similar Response ◽  
Tertiary Amines ◽  
Microbial Oxidation ◽  
Ph Optimum ◽  
Tetramethylammonium Chloride

1. A mono-oxygenase, which oxidizes trimethylamine and other tertiary amines bearing methyl or ethyl groups, was partially purified sixfold from Pseudomonas aminovorans grown on trimethylamine as sole carbon source. 2. The preferred electron donor was NADPH. The enzyme had a pH optimum of 8.0–9.4 for trimethylamine oxidation, and 8.8–9.2 for dimethylamine oxidation. 3. The oxidation product of trimethylamine was shown to be trimethylamine N-oxide. Other tertiary amines were probably also converted into N-oxides. 4. The enzyme also oxidized secondary amines. 5. The oxidation of trimethylamine was only slightly inhibited by CO and not at all by KCN or proadifen hydrochloride (SKF 525-A), but was inhibited by trimethylsulphonium chloride, tetramethylammonium chloride, 2,4-dichloro-6-phenylphenoxyethylamine (Lilly 53325) and its NN-diethyl derivative (Lilly 18947). 6. The oxidation of dimethylamine showed a similar response to inhibitors and a parallel loss in activity on heating at 35°C. 7. The activities of the trimethylamine mono-oxygenase, trimethylamine N-oxide demethylase and the secondary-amine mono-oxygenase increased severalfold during adaptation of succinate-grown bacteria to growth on trimethylamine, and the trimethylamine mono-oxygenase was the first enzyme to show an increase in activity. It is concluded that all three enzymes are involved in growth on trimethylamine by this organism.

scholarly journals Biodecolourisation of Reactive Red 120 as a Sole Carbon Source by a Bacterial Consortium—Toxicity Assessment and Statistical Optimisation

2021 ◽  
Vol 18 (5) ◽  
pp. 2424
Author(s):  
Motharasan Manogaran ◽  
Nur Adeela Yasid ◽  
Ahmad Razi Othman ◽  
Baskaran Gunasekaran ◽  
Mohd Izuan Effendi Halmi ◽  
...  
Keyword(s):  
Carbon Source ◽  
Ammonium Sulphate ◽  
Azo Dye ◽  
Sole Carbon Source ◽  
Bacterial Consortium ◽  
Toxicity Assessment ◽  
Substrate Availability ◽  
Reactive Red 120 ◽  
Significant Attention

The application of microorganisms in azo dye remediation has gained significant attention, leading to various published studies reporting different methods for obtaining the best dye decolouriser. This paper investigates and compares the role of methods and media used in obtaining a bacterial consortium capable of decolourising azo dye as the sole carbon source, which is extremely rare to find. It was demonstrated that a prolonged acclimation under low substrate availability successfully isolated a novel consortium capable of utilising Reactive Red 120 dye as a sole carbon source in aerobic conditions. This consortium, known as JR3, consists of Pseudomonas aeruginosa strain MM01, Enterobacter sp. strain MM05 and Serratia marcescens strain MM06. Decolourised metabolites of consortium JR3 showed an improvement in mung bean’s seed germination and shoot and root length. One-factor-at-time optimisation characterisation showed maximal of 82.9% decolourisation at 0.7 g/L ammonium sulphate, pH 8, 35 °C, and RR120 concentrations of 200 ppm. Decolourisation modelling utilising response surface methodology (RSM) successfully improved decolourisation even more. RSM resulted in maximal decolourisation of 92.79% using 0.645 g/L ammonium sulphate, pH 8.29, 34.5 °C and 200 ppm RR120.

scholarly journals Studies on mammalian glucoamylases with special reference to monkey intestinal glucoamylase

1970 ◽  
Vol 117 (5) ◽  
pp. 939-946 ◽  
Author(s):  
B. Seetharam ◽  
N. Swaminathan ◽  
A. N. Radhakrishnan
Keyword(s):  
Special Reference ◽  
Enrichment Factor ◽  
Intestinal Mucosa ◽  
Liver Enzymes ◽  
Gel Filtration ◽  
Maximum Activity ◽  
Similar Magnitude ◽  
Glucoamylase Activity ◽  
Specific Activities ◽  
Intestinal Enzyme

1. Highly purified preparations of glucoamylase were obtained from liver, spleen and intestine of the monkey. The enrichment factor was lower for intestine (60-fold) compared with that of liver (1200-fold) and of spleen (2000-fold) but the final specific activities were of a similar magnitude. 2. The liver and spleen enzymes had maximum activity at pH4.8 whereas the intestinal enzyme showed an optimum at pH5.8. The Km values for both starch and maltose with spleen and liver enzymes were higher than for the intestinal enzyme. With the intestinal enzyme, the Vmax. values were higher for both starch and maltose than those of the spleen and liver enzymes. 3. Gel filtration on Sephadex G-200 under identical conditions revealed that liver and spleen enzymes emerge from the columns much later than the intestinal enzyme. 4. Evidence is presented that the glucoamylase activity of the intestinal mucosa is exhibited by the maltase II fraction. 5. Tris, pentaerythritol and turanose inhibited glucoamylase from all the three tissues, but turanose inhibited the spleen and liver enzymes to a higher degree than the intestinal enzyme.

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