scholarly journals Purification and characterization of the invertase from Schizosaccharomyces pombe. A comparative analysis with the invertase from Saccharomyces cerevisiae

1990 ◽  
Vol 267 (3) ◽  
pp. 697-702 ◽  
Author(s):  
S Moreno ◽  
Y Sanchez ◽  
L Rodriguez
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Schizosaccharomyces Pombe ◽  
Molecular Mass ◽  
Total Mass ◽  
High Molecular Mass ◽  
Carbohydrate Moiety ◽  
Purification And Characterization ◽  
Protein Moiety ◽  
Protein Portion

Invertase (EC 3.2.1.26) was purified to homogeneity from exponentially growing cells of Schizosaccharomyces pombe fully de-repressed for synthesis of the enzyme, and was shown to be a high-molecular-mass glycoprotein that can be dissociated in the presence of 8 M-urea/1% SDS into identical subunits with an apparent molecular mass of 205 kDa. The carbohydrate moiety, accounting for 67% of the total mass, is composed of equimolar amounts of mannose and galactose. There is a small amount of glucosamine, which is probably involved in the linkage to the protein moiety, since the enzyme is sensitive to treatment with endoglycosidase H. The composition of the carbohydrate moiety resembles that found in higher-eukaryotic glycoproteins and differs from glycoproteins found in Saccharomyces cerevisiae. The protein portion of each subunit is a polypeptide of molecular mass 60 kDa, very similar to the invertase of Sacch. cerevisiae. Both proteins cross-react with antibodies raised against the protein fractions of the other, indicating that the two enzymes are similar.

Purification and characterization of fructans with β‐2, 1‐ and β‐2, 6‐glycosidic linkages suitable for enzyme studies

2020 ◽  
Author(s):  
GD BONNETT ◽  
Ian Sims ◽  
JA ST. JOHN ◽  
RJ SIMPSON
Keyword(s):  
Molecular Mass ◽  
Small Proportion ◽  
Gel Filtration ◽  
Triticum Aestivum L ◽  
High Molecular Mass ◽  
Enzyme Assays ◽  
Purification And Characterization ◽  
Linear Molecules ◽  
Low Levels

Fructan pentasaccharides were purified, in quantities suitable for use as substrates for enzyme assays, from Neosugar‐p‐(Meijj Seika Kaisha Ltd. Japan), tubers of Helianthus tuberosus L., L., and stems and leaf sheaths of Triticum aestivum L by a combination of gel‐filtration and RP‐HPLC. Fructan of higher molecular mass (mean DP = 30) was purified from Leaves of Lolium rigidum Gaud, that had been induced to accumulate fructan and characterized along; with the commercially available fructan from Cichorium intybus L. (Sigma, St Louis, USA) (mean DP = 33). The fructan pentasaccharide purified from H. tuberosus was found to contain exclusively 2, 1‐linked fructose and terminal fructose and terminal glucose, and was identified as (1, 1, 1)‐kestopentatise. The fructan pentasaccharide purified from Neosugar‐P also contained (1,1,1)‐kestopentaose. although the presence of fructan Klinked glucose and 1 % 2, 6‐linked fructose indicated that a small proportion of other kestopentaoses were present, The fructan pentasaccharide purified from T aestivum consisted of almost exclusively 2,6‐linked fructose and terminal glucose and terminal fructose and was considered to contain predominantly (6,6,6)‐kestopentaose. The presence of 1 % 2,1,6)‐linked fructose indicated the sample also contained a small proportion of branched kestopentanse. The high molecular mass fructan from C. intybus was found to comprise linear molecules containing only 2,1‐linked fructose, terminal glucose and terminal fructose‐ High molecular mass fructan from L. rigidum contained predominantly 2. h‐linked fructose, had predominantly internal glucose, indicated by 2 %, 1.6‐linked glucose, low levels of branching, indicated 2 % 2,1,6‐linked fructose residues; and 1% of the residues were 2,1 ‐linked fructose. Copyright © 1994, Wiley Blackwell. All rights reserved

scholarly journals Characterization of the invertase from Pichia anomala

1995 ◽  
Vol 306 (1) ◽  
pp. 235-239 ◽  
Author(s):  
J Rodriguez ◽  
J A Perez ◽  
T Ruiz ◽  
L Rodriguez
Keyword(s):  
Carbon Source ◽  
Molecular Mass ◽  
Total Mass ◽  
Culture Medium ◽  
Thermal Sensitivity ◽  
Apparent Molecular Mass ◽  
Carbohydrate Moiety ◽  
Pichia Anomala

Synthesis of invertase (EC 3.2.1.26) in Pichia anomala is controlled by the carbon source in the culture medium. The enzyme was purified to homogeneity from P. anomala cells fully derepressed for invertase synthesis and shown to be a multimeric glycoprotein composed of identical subunits with an apparent molecular mass of 86.5 kDa. The carbohydrate moiety accounts for approx. 30% of the total mass of the molecule and consists of manno-oligosaccharides N-linked to the polypeptide. Most of the characteristics of the enzyme analysed in this study were similar to those previously reported for other yeast invertases, with the remarkable exception of its thermal sensitivity which appears after 15 min incubation at temperatures above 32 degrees C.

Purification and characterization of fructans with β‐2, 1‐ and β‐2, 6‐glycosidic linkages suitable for enzyme studies

2020 ◽  
Author(s):  
GD BONNETT ◽  
Ian Sims ◽  
JA ST. JOHN ◽  
RJ SIMPSON
Keyword(s):  
Molecular Mass ◽  
Small Proportion ◽  
Gel Filtration ◽  
Triticum Aestivum L ◽  
High Molecular Mass ◽  
Enzyme Assays ◽  
Purification And Characterization ◽  
Linear Molecules ◽  
Low Levels

Fructan pentasaccharides were purified, in quantities suitable for use as substrates for enzyme assays, from Neosugar‐p‐(Meijj Seika Kaisha Ltd. Japan), tubers of Helianthus tuberosus L., L., and stems and leaf sheaths of Triticum aestivum L by a combination of gel‐filtration and RP‐HPLC. Fructan of higher molecular mass (mean DP = 30) was purified from Leaves of Lolium rigidum Gaud, that had been induced to accumulate fructan and characterized along; with the commercially available fructan from Cichorium intybus L. (Sigma, St Louis, USA) (mean DP = 33). The fructan pentasaccharide purified from H. tuberosus was found to contain exclusively 2, 1‐linked fructose and terminal fructose and terminal glucose, and was identified as (1, 1, 1)‐kestopentatise. The fructan pentasaccharide purified from Neosugar‐P also contained (1,1,1)‐kestopentaose. although the presence of fructan Klinked glucose and 1 % 2, 6‐linked fructose indicated that a small proportion of other kestopentaoses were present, The fructan pentasaccharide purified from T aestivum consisted of almost exclusively 2,6‐linked fructose and terminal glucose and terminal fructose and was considered to contain predominantly (6,6,6)‐kestopentaose. The presence of 1 % 2,1,6)‐linked fructose indicated the sample also contained a small proportion of branched kestopentanse. The high molecular mass fructan from C. intybus was found to comprise linear molecules containing only 2,1‐linked fructose, terminal glucose and terminal fructose‐ High molecular mass fructan from L. rigidum contained predominantly 2. h‐linked fructose, had predominantly internal glucose, indicated by 2 %, 1.6‐linked glucose, low levels of branching, indicated 2 % 2,1,6‐linked fructose residues; and 1% of the residues were 2,1 ‐linked fructose. Copyright © 1994, Wiley Blackwell. All rights reserved

Purification and characterization of a high molecular mass serine carboxypeptidase from Monascus pilosus

2004 ◽  
Vol 31 (12) ◽  
pp. 572-580 ◽  
Author(s):  
Fang Liu ◽  
Shinjiro Tachibana ◽  
Toki Taira ◽  
Masanobu Ishihara ◽  
Fumio Kato ◽  
...  
Keyword(s):  
Molecular Mass ◽  
High Molecular Mass ◽  
Serine Carboxypeptidase ◽  
Purification And Characterization ◽  
Monascus Pilosus
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