scholarly journals The specificity of proteinases from Streptomyces griseus (pronase)

1970 ◽  
Vol 116 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Moshe Trop ◽  
Yehudith Birk
Keyword(s):  
Streptomyces Griseus ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Considerable Extent ◽  
Small Peptides ◽  
Diisopropyl Phosphorofluoridate ◽  
Naturally Occurring ◽  
Bovine Trypsin ◽  
Wide Range ◽  
Pancreatic Trypsin Inhibitor

Purification of pronase by ion-exchange chromatography gave four proteolytically active fractions. Fraction A2 contained an endopeptidase that attacks poly l-valine. Fraction B contained an endopeptidase, an aminopeptidase and carboxypeptidases. The activities against hippuryl-L-arginine and hippuryl-L-phenylalanine could be inhibited to a considerable extent by di-isopropyl phosphorofluoridate and by EDTA. Fraction C contained an endopeptidase resembling bovine trypsin. The pure enzyme was completely inactivated by di-isopropyl phosphorofluoridate and pancreatic trypsin inhibitor and to about 90% by other naturally occurring trypsin inhibitors. Fraction D contained an apparently homogeneous endopeptidase, inhibited by diisopropyl phosphorofluoridate, that adsorbed to and hydrolysed elastin. The activity of all these fractions was tested qualitatively against a wide range of small peptides and synthetic substrates.

scholarly journals Antigenicity of proteinases from Streptomyces griseus (pronase)

1970 ◽  
Vol 119 (2) ◽  
pp. 339-342 ◽  
Author(s):  
M. Trop ◽  
R. R. Avtalion ◽  
Z. Malik ◽  
A. Pinsky
Keyword(s):  
Amino Acid ◽  
Active Sites ◽  
Streptomyces Griseus ◽  
Ion Exchange Chromatography ◽  
Enzymic Activity ◽  
Exchange Chromatography ◽  
Soya Bean ◽  
Catalytic Site ◽  
Antigenic Sites ◽  
Bovine Trypsin

The five pronase fractions, A1, A2, B, C (trypsin-like), and D (elastolytic), obtained by ion-exchange chromatography, were found to be antigenically distinct. Antibodies to pronase inhibited the enzymic activity of each of the enzyme fractions. Pronase trypsin and bovine trypsin, although resembling each other in enzymic activity and in amino acid sequence around their active sites, did not cross-react antigenically with, nor was their enzymic activity inhibited by, the respective homologous antibodies. Inactivation of pronase trypsin by complexing with soya-bean inhibitor AA, was not associated with a decrease in capacity to precipitate with its antibody. It is assumed that the antigenic sites are located far enough from the catalytic site of the enzyme to allow it to precipitate immunologically even when the catalytic site was blocked.

Disulphide arrangement in bovine caseins: localization of intrachain disulphide bridges in monomers of κ- and αs2-casein from bovine milk

1994 ◽  
Vol 61 (4) ◽  
pp. 485-493 ◽  
Author(s):  
Lone K. Rasmussen ◽  
Peter Højrup ◽  
Torben E. Petersen
Keyword(s):  
Mass Spectrometry ◽  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Bovine Milk ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequence Analysis ◽  
Naturally Occurring ◽  
Localize Potential

SummaryNaturally occurring monomeric κ-casein and αs2-casein in bovine milk were purified by ion-exchange chromatography in order to localize potential intrachain disulphide bridges. Enzymic cleavage of the proteins followed by mass spectrometry and amino acid sequence analysis of cystine-containing peptides revealed the presence of an intrachain disulphide bond in both proteins.

scholarly journals Industry Perspectives on the Use of Natural Antimicrobials and Inhibitors for Food Applications

1996 ◽  
Vol 59 (13) ◽  
pp. 82-86 ◽  
Author(s):  
GRAHAME W. GOULD
Keyword(s):  
Food Preservation ◽  
Small Peptides ◽  
Natural Systems ◽  
Naturally Occurring ◽  
Wide Range ◽  
Food Applications ◽  
Future Potential ◽  
Product Quality And Safety ◽  
Natural Inhibitors ◽  
Synergistic Combinations

ABSTRACT The wide range of extremely effective naturally occurring antimicrobial systems include those derived front animals (e.g., enzymes such as lysozyme and lactoperoxidase; other proteins such as lactoferrin, lactoferricin, ovotransferrin, and serum transferrins; small peptides such as histatins and magainins; and the immune system), those derived front plants (e.g., phytoalexins, low- molecular-weight components of herbs and spices; phenolics such as oleuropein; and essential oils) and those derived front microorganisms (e.g., bacteriocins such as nisin and pediocin). An increasing number of such natural systems is being deliberately utilized for food preservation, or being explored for such use. The future potential is substantial, particularly as the efficacy of these systems is demonstrated in additive or synergistic combinations with some of the other antimicrobial factors that we can employ to improve the safety and shelf stability of foods. While “naturalness” alone is not necessarily a sufficient objective for these developments, the use of natural inhibitors as components of systems that can together enhance the effectiveness of preservation, with advantages in product quality and safety, justifies pursuit.

scholarly journals Production of amylases by Aspergillus tamarii

1999 ◽  
Vol 30 (2) ◽  
pp. 157-162 ◽  
Author(s):  
Fabiana Guillen Moreira ◽  
Francieli Arrias de Lima ◽  
Sophia Renata Fazzano Pedrinho ◽  
Veridiana Lenartovicz ◽  
Cristina Giatti Marques de Souza ◽  
...  
Keyword(s):  
Carbon Source ◽  
Ion Exchange ◽  
Filamentous Fungus ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Aspergillus Tamarii ◽  
Ph Values ◽  
Wide Range ◽  
Initial Culture ◽  
Static Cultivation

A strain of Aspergillus tamarii, a filamentous fungus isolated from soil, was able to produce both <FONT FACE="Symbol">a</FONT>-amylase and glucoamylase activities in mineral media supplemented with 1% (w/v) starch or maltose as the carbon source. Static cultivation led to significantly higher yields than those obtained using shaking culture. The production of amylases was tolerant to a wide range of initial culture pH values (from 4 to 10) and temperature (from 25 to 42oC). Two amylases, one <FONT FACE="Symbol">a</FONT>-amylase and one glucoamylase, were separated by ion exchange chromatography. Both partially purified enzymes had optimal activities at pH values between 4.5 and 6.0 and were stable under acid conditions (pH 4.0-7.0). The enzymes exhibited optimal activities at temperatures between 50o and 60o C and were stable for more than ten hours at 55oC.

IODOPEPTIDES FROM HUMAN THYROGLOBULIN

1977 ◽  
Vol 85 (4) ◽  
pp. 769-780 ◽  
Author(s):  
Lubomir J. Valenta ◽  
A. Donny Strosberg ◽  
Vera Valenta ◽  
Jean-Claude Jaton
Keyword(s):  
Ion Exchange ◽  
Peptide Mapping ◽  
Thyroid Tissue ◽  
Iodine Content ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Small Peptides ◽  
Thyroid Glands ◽  
High Voltage Electrophoresis

ABSTRACT Human thyroglobulin labelled in vivo by 125I was purified from eight different thyroid glands including normal thyroid tissue, thyrotoxic goitre and euthyroid multinodular goitre. The purified protein was cleaved with cyanogen bromide (CNBr) and the resulting peptides were separated by column chromatography and ion exchange chromatography. Reproducible elution profiles of both protein and iodine were obtained. However, the distribution of iodine depended on the iodine content of the intact thyroglobulin. Small CNBr peptides seemed to be preferentially iodinated, but with a limited capacity. With higher degrees of iodination, larger peptides became richer in iodine. This suggests sequential iodination of the thyroglobulin molecule. The mixture of small peptides was digested by trypsin. Two iodopeptides were identified in this material by peptide mapping and they had identical migration in thyroglobulins of different origin. One of them was purified by ion exchange chromatography and high voltage electrophoresis. Analogous amino acid composition was obtained for the iodopeptide purified from two different thyroglobulins. The data indicates that thyroglobulin iodination occurs in specific portions of the polypeptide chain and probably in a sequential manner.

The Amino Acid Sequence of Streptomyces griseus Trypsin. I. The Peptic Peptides

1973 ◽  
Vol 51 (7) ◽  
pp. 1077-1088 ◽  
Author(s):  
L. Jurášek ◽  
L. B. Smillie
Keyword(s):  
Amino Acid ◽  
Serine Proteases ◽  
System Analysis ◽  
Streptomyces Griseus ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Terminal Sequence ◽  
Unique Amino Acid ◽  
High Voltage Electrophoresis

Streptomyces griseus trypsin (S.G.T.) isolated from pronase was digested with pepsin. The peptic peptides were isolated by high-voltage electrophoresis on paper and ion-exchange chromatography on Chromobead P resin using the Technicon autoanalyzer system. Analysis of the purified peptides provides 28 unique amino acid sequences accounting for approximately 95% of the S.G.T. molecule. A portion of the residues not accounted for can be ascribed to free leucine, phenylalanine, tyrosine, and tryptophan present in the peptic digest. The NH2-terminal sequence of S.G.T. was shown to be Val–Val–Gly–Gly–Thr–Arg–Ala–Ala–Gln–Gly–Glu–Phe and is highly homologous with NH2-terminal sequences of other Asp–Ser–Gly serine proteases.

The Amino Acid Sequence of Streptomyces griseus Trypsin. II The Tryptic Peptides

1974 ◽  
Vol 52 (5) ◽  
pp. 382-392 ◽  
Author(s):  
L. Jurášek ◽  
L. B. Smillie
Keyword(s):  
Soluble Fraction ◽  
Streptomyces Griseus ◽  
Paper Electrophoresis ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Insoluble Residue ◽  
Limited Extent ◽  
Terminal Sequence ◽  
Tryptic Peptides ◽  
Final Purification

Streptomyces griseus trypsin (S.G.T.) isolated from pronase was reduced, aminoethylated, and digested with trypsin. The soluble peptides were recovered and the insoluble residue redigested with chymotrypsin. Following recovery of the soluble fraction, the insoluble portion was in turn digested with α-lytic protease of Myxobacter 495. The three groups of soluble peptides were separately subjected to ion-exchange chromatography on the Technicon peptide analyzer and to final purification by high-voltage paper electrophoresis. Sequence analysis by the dansyl-Edman procedure provided unique sequences from the soluble tryptic peptides accounting for 65% of the S.G.T. molecule. Peptides obtained from the redigests of the insoluble residues accounted for an additional 20%.Tryptic digestion of dansylated S.G.T. yielded a unique α-NH2 dansylated peptide whose composition showed it to be the same as the NH2-terminal sequence previously postulated for this enzyme. The tryptic peptides isolated in this work have provided overlaps for many of the previously sequenced peptic peptides. Three continuous sequences of 49, 36, and 28 residues have been elucidated.Evidence has also been obtained that the NH2-terminal valine residue was, to a limited extent, aminoethylated during reaction of the reduced protein with ethylenimine.

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