Purification of extracellular amylolytic enzymes from the thermophilic fungus Thermomyces lanuginosus

1988 ◽  
Vol 34 (3) ◽  
pp. 218-223 ◽  
Author(s):  
Bo Jensen ◽  
Jorgen Olsen ◽  
Knud Allermann
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Soluble Starch ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Thermomyces Lanuginosus ◽  
Amylolytic Enzymes ◽  
Molecular Weights ◽  
Isoelectric Points ◽  
Hydrolysis Of

When grown in static culture it appears as if Thermomyces lanuginosus has a biphasic secretion of the extracellular starch-degrading activity. This could be due to the presence of at least two different amylases. By ion-exchange chromatography on DEAE-Trisacryl an α-amylase (EC 3.2.1.1) and a glucoamylase (EC 3.2.1.3) were separated and purified from the extracellular protein from 14-day-old static cultures grown on soluble starch. The hydrolysis of soluble starch by the purified glucoamylase resulted in only glucose as the end product, whereas the α-amylase gave maltose as the smallest end product. The molecular weights and isoelectric points of the enzymes were for glucoamylase 70 000 – 76 000 and pH 4.0, and for α-amylase 54 000 – 57 000 and pH 3.4. An α-glucosidase (EC 3.2.1.20) with a molecular weight of 44 000 – 48 000 and an isoelectric point at pH 3.8 was eluted close to the α-amylase fraction on the DEAE-Trisacryl column.

Some properties of a highly thermostable α-amylase from a Thermoactinomyces sp.

1984 ◽  
Vol 30 (6) ◽  
pp. 780-785 ◽  
Author(s):  
S. K. C. Obi ◽  
F. J. C. Odibo
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Ion Exchange ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzyme Activation ◽  
Cow Dung ◽  
C 30 ◽  
Hydrolysis Of

Thermostable α-amylase from Thermoactinomyces sp. No. 15, isolated from cow dung, was partially purified and characterized. The enzyme was purified (318-fold) by acetone precipitation, ion-exchange chromatography, and gel filtration techniques. The molecular weight was estimated to be 47 800. Optimum enzyme activity was recorded at pH 7 and at 80 °C. The enzyme was stable at pH 5.0–10.0 and retained 74% activity at 100 °C (30 min). Enzyme activation was observed in the presence of Mn2+, Ag+, and Fe2+, but Hg2+ and Zn2+ were inhibitory. Products of hydrolysis of native starches were mainly glucose and maltose.

scholarly journals Rat small-intestinal β-galactosidases. Studies on the fractionation of ‘acid’ β-galactosidase with isoelectric focusing, gel filtration and ion-exchange chromatography

1970 ◽  
Vol 117 (2) ◽  
pp. 369-375 ◽  
Author(s):  
Nils-Georg Asp
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
High Molecular Weight ◽  
Isoelectric Focusing ◽  
Gel Filtration ◽  
Ion Exchange Resin ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Isoelectric Points ◽  
Small Intestinal

1. Different forms of the rat small-intestinal ‘acid’ β-galactosidase were separated by using the isoelectric-focusing technique. The isoelectric points of the different forms were at pH4.2, 4.6, 5.4, 6.1 and approx. 8. 2. The two forms of ‘acid’ β-galactosidase isoelectric at pH4.2 and 4.6 were completely excluded from the Sephadex G-200 gel, whereas the form isoelectric at pH8 had Kav. 0.4. The concentration and pH of the elution buffer influenced the distribution of enzyme activity between different forms. Thus, under certain conditions of ionic strength and pH, the enzyme seems to form high-molecular-weight aggregates with low isoelectric points. These may be homopolymeric aggregates or the result of binding of enzyme to, for example, membrane fragments. The forms isoelectric at pH5.4 and 6.1 are probably aggregates of intermediate size. 3. During ion-exchange chromatography at pH6.0 one fraction of ‘acid’ β-galactosidase was not retained on the column and was isoelectric at pH8 and another fraction was eluted when the buffer concentration in the eluate had increased to about 50mm. The main part of enzyme eluted in this second fraction was also isoelectric at pH8, indicating that the elution of this fraction is not a simple ion-exchange procedure but probably also involves a splitting of high-molecular-weight aggregates, originally retained because of their low isoelectric points. The enzyme subunits have a higher isoelectric point, and are therefore no longer bound to the ion-exchange resin.

scholarly journals Access to N-Acetylated Chitohexaose with Well-Defined Degrees of Acetylation

2017 ◽  
Vol 2017 ◽  
pp. 1-15 ◽  
Author(s):  
Kecheng Li ◽  
Ronge Xing ◽  
Song Liu ◽  
Yukun Qin ◽  
Pengcheng Li
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Esi Ms ◽  
Molecular Weights

Chitohexaose has attracted wide interest due to its special bioactivities and these potential activities are significantly related to N-acetylation. Herein, six chitohexaose fractions with different degrees of acetylation were prepared by selective N-acetylation and ion-exchange chromatography and further analyzed by ESI/MS. It is revealed that all the six N-acetylated chitohexaoses were of single molecular weight, the molecular weights of which were exactly assigned to 1026.44 Da, 1068.44 Da, 1110.48 Da, 1152.48 Da, 1194.49 Da, and 1236.48 Da, respectively. These results suggested that the six prepared N-acetylated chitohexaoses were N-acetylchitohexaose (D5A1), di-N-acetylchitohexaose (D4A2), tri-N-acetylchitohexaose (D3A3), tetra-N-acetylchitohexaose (D2A4), penta-N-acetylchitohexaose (D1A5), and hexa-N-acetylchitohexaose (A6), respectively, which are of great significance to screen their bioactivities and discover well-defined chitooligosaccharide molecules as potential drugs.

PURIFICATION AND CHARACTERISATION OF FIBRINOLYTIC ENZYMES FROM ENDOPHYTIC FUNGI AND LIGNOSUS RHINOCERUS

2016 ◽  
Vol 78 (6-5) ◽  
Author(s):  
Zainon Mohd Noor ◽  
Mohd Sidek Ahmad ◽  
Zaidah Zainal Ariffin
Keyword(s):  
Ion Exchange ◽  
Endophytic Fungi ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fibrinolytic Enzymes ◽  
Molecular Weights ◽  
Ammonium Sulphate Precipitation ◽  
Fusarium Sp ◽  
Different Sources

Three enzymes FH3, S13 and LR1 from three different sources showed fibrinolytic activities. Two were from endophytic fungal cultures and one from the sclerotium of Lignosus rhinocerus mushroom (LR1). FH3, S13 cultures and LR1, the crude extract of the sclerotium were concentrated and purified by ammonium sulphate precipitation, ion-exchange chromatography and gel-filtration. The molecular weights of the FH3, S13 and LR1 purified enzymes were estimated to be approximately 34kDa, 34kDa and 10kDa, respectively. Maximum fibrinolytic activities were observed for FH3 at pH 7 and 30°C, S13 at pH 8 and 40°C and LR1 at pH 6 and 40°C.  In our earlier paper we identified FH3 as Fusarium sp. and S13 as Penicilium citrinum. 

A dipeptidylpeptidase secreted by Fasciola hepatica

Parasitology ◽  
1994 ◽  
Vol 109 (1) ◽  
pp. 113-118 ◽  
Author(s):  
C. Carmona ◽  
S. McGonigle ◽  
A. J. Dowd ◽  
A. M. Smith ◽  
S. Coughlan ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Liver Fluke ◽  
Fasciola Hepatica ◽  
Gel Filtration ◽  
Serine Proteinase ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Substrate Preference ◽  
Proteolytic Digestion

SUMMARYA dipeptidylpeptidase (DPP) was isolated from Fasciola hepatica by gel-filtration and ion-exchange chromatography. The exoproteinase is secreted by newly excysted juveniles, immature and mature flukes. The liver fluke DPP is a serine proteinase of molecular weight > 200 kDa and differs from previously characterized mammalian DPPs in its substrate preference and susceptibility to inactivation by inhibitors. The parasite DPP may function in the latter stages of the proteolytic digestion of host macromolecules. In this manner, the enzyme may be important in providing the parasite with dipeptides that could be absorbed through the intestine as nutrient.

scholarly journals A Polysaccharide Produced by Lactococcus lactis subsp. lactis YZ1 Isolated from Traditional Indonesian Fermented Milk, "Dadih"

2000 ◽  
Vol 1 (1) ◽  
pp. 1-5
Author(s):  
Yusdar Zakaria
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Lactococcus Lactis ◽  
Monosaccharide Composition ◽  
Fermented Milk ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Molar Ratio ◽  
Neutral Sugar ◽  
Glycoside Linkage

ABSTRACT.Lactococcus lactis subsp. Lactis YZI was isolated from M17 agar in which diluted Dadih was poured and incubated at 30 0C for 48 h. Taxonomix properties of the isolate were examined according to Bergey’s Manual of Systematic Bacteriologi and Manual for  Identification of Medical Bacteria. The isolation of polysaccharide from the precipitant was performed on an ion-exchange chromatography. The result showed that the polysaccharides produced by Lactococus lactis subsp. lactis YZI were neutral sugar (unadsorbrd fraction) and glycoconjugated (absorbed fraction). The neutral sugar had molecular weight of 10,000 and 20,000 with and α-glycoside linkage. The monosaccharide composition was mannose, glucose and galactose with a molar ratio of 1 :1,5 : 4,9.

scholarly journals High-Sulphur Proteins From a-Keratins ll. Isolation and Partial Characterization of Purified Components From Mouse Hair

1976 ◽  
Vol 29 (2) ◽  
pp. 11 ◽  
Author(s):  
Robert C Marshall ◽  
JM Gillespie
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fractional Precipitation ◽  
Molecular Weight Range

The present paper continues the study of the reduced and S-carboxymethylated high-sulphur proteins from mouse hair. Fractions have been obtained in a substantially purified form by fractional precipitation with ammonium sulphate at pH 6, followed by ion exchange chromatography on cellulose phosphate at pH 2�6. Approximately 80% by weight of the high-sulphur proteins fall into the ultra-high-sulphur category (carboxymethyicysteine content greater than 26 residues per 100 residues), and they cover a molecular weight range of 17000-28000. The components show a remarkable diversity in amino acid composition; for example the contents of arginine and glycine each vary by about 3 : 1. The remainder of the proteins contain 17-20 residues per 100 residues of carboxymethyicysteine, are smaller in size (molecular weight 11 500), and also show great diversity in overall amino acid composition.

scholarly journals PEMISAHAN ASAM AMINO DARI LIMBAH CAIR PABRIK KELAPA SAWIT DENGAN KROMATOGRAFI PENUKAR ION

2018 ◽  
Vol 6 (2) ◽  
pp. 69
Author(s):  
Indah Rahmi Sari ◽  
Ade Ayu Oksari ◽  
Irma Kresnawaty
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Liquid Waste ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzyme Hydrolysis ◽  
Protease Enzyme ◽  
Absorbance Reading ◽  
Amino Acid Levels ◽  
Hydrolysis Of

Separation of Amino acid from Liquid waste of Oil palm Factory with Ion Exchange Chromatography Research on Separation of Amino Acid Liquid Waste mills with Ion Exchange Chromatography was carried out from October to November 2015. The results of  hydrolysis of 6 N HCl results showed  that the highest absorbance reading was obtained at a concentration of eluent of 0,2 and 0,6 M NaCl, while the results of the protease enzyme hydrolysis the highest absorbance reading at NaCl eluent 0,2 and 1 M. There was no significant difference in the results of separation by ion exchange chromatography, showed that the concentration of NaCl eluent is not very influential, so for subsequent analysis used only one concentration of the eluent. Results of linear regression obtained was equal to 0,9946, these results indicate that the series standard amino acid lysine has a value that is linear as it approaches 1. The amino acid levels obtained on the sample results LCPKS hydrolysis with 6 N HCl which was about 0 to 8.82 ppm and samples of the protease enzyme hydrolysis of about 0 to 4.31 ppm. Amino acid levels obtained were still far from the expected.Keywords: Amino Acid, Oil Palm, Liquid Waste, Ion Exchange Chromatography ABSTRAKPenelitian mengenai Pemisahan Asam Amino dari Limbah Cair Pabrik Kelapa Sawit dengan Kromatografi Penukar Ion telah dilaksanakan dari bulan Oktober sampai November 2015. Hasil hidrolisis HCl 6 N menunjukkan bahwa pembacaan absorbansi tertinggi diperoleh pada konsentrasi eluen NaCl 0,2 dan 0,6 M, sedangkan hasil hidrolisis enzim protease pembacaan absorbansi tertinggi pada eluen NaCl 0,2 dan 1 M. Tidak ada perbedaan yang signifikan pada hasil pemisahan dengan kromatografi penukar ion ini, menunjukkan bahwa konsentrasi eluen NaCl tidak terlalu berpengaruh, sehingga untuk analisis selanjutnya digunakan hanya satu konsentrasi eluen. Hasil regresi linear yang diperoleh yaitu sebesar 0,9946, hasil tersebut menunjukkan bahwa deret standar asam amino lisin mempunyai nilai yang linear karena mendekati 1. Kadar asam amino yang diperoleh pada sampel hasil hidrolisis LCPKS dengan HCl 6N yaitu sekitar 0 – 8,82 ppm dan sampel hasil hidrolisis enzim protease sekitar 0 – 4,31 ppm. Kadar asam amino yang diperoleh masih jauh dari yang diharapkan.Kata Kunci: Asam Amino, Minyak Kelapa Sawit, Limbah Cair, Kromatografi Penukar Ion

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