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Extraction, partial purification and characterization of tomato invertase

TL;DR: The enzyme invertase was isolated and partially purifi ed from healthy tomato juice and was found to be stable at the temperature ranged from 10oC to 35oC, and the K m value of this enzyme for sucrose was 4.5 mM at pH 5.2.
Abstract: The enzyme invertase (β-D-fructofuranoside fructohydrolase; E.C. 3.2.1.26) was isolated and partially purifi ed from healthy tomato juice. The purifi cation involved buffer extraction, DEAE- cellulose and Sephadex G-75 chromatography. The purity of the enzyme preparation was determined by SDS-PAGE. The enzyme was purifi ed 29.9 fold with 23.19% yield, giving a fi nal specifi c activity 87.62 U/mg. The molecular weights of the purifi ed enzymes measured by gel fi ltration chromatogra- phy and SDS-PAGE were found to be 54 kDa and 49 kDa, respectively. The purifi ed invertase was a glycoprotein with 17.5% sugar. The optimum pH of the purifi ed enzyme was 5.5 and the activity was stable at pH 3.5-7.5. The enzyme showed maximum activity at 35oC and was found to be stable at the temperature ranged from 10oC to 35oC. The K m value of this enzyme for sucrose was 4.5 mM at pH 5. Tris, glucose and fructose reduced invertase activities poorly while urea, EDTA, acetic acid, Zn 2+ and Cd 2 + decreased moderately. Ag+ and Al 3 + produced a slight inhibitory effect on invertase activity. Ca 2+ had almost no effect on tomato invertase activity. Mn 2+ , Mg 2+ , K+, Na+ and Ba 2+ increased invertase activity slightly, while Cu 2+ accelerate invertase action moderately. Hg 2+ almost completely ceased the
Citations
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Journal ArticleDOI
TL;DR: In this paper, the authors extract and purify tomato (Lycopersicon esculentum) invertase using an aqueous two-phase system (ATPS), which is formed by mixing the polymer with a salt and a protein solution.

77 citations

Journal ArticleDOI
TL;DR: Three-phase partitioning, a technique used in protein purification, was used to purify invertase from tomato and shows that, TPP is a simple, quick and economical technique for purification of invertases.

65 citations

Journal ArticleDOI
TL;DR: Invertase belonging to glycoside hydrolases that catalyzes sucrose (table sugar) into two monosaccharides i.e. glucose and fructose assist in defense reaction against bacterial infections and gut fermentation by oxidation and categorize them as powerful antimicrobial and antioxidant agents.

63 citations

Journal ArticleDOI
TL;DR: This paper provides comprehensive information on the invertase and their types, sources, production methods, physical and chemical properties, structures, separation and purification methods, and their applications.
Abstract: Invertase (beta-fructofuranosidase) enzyme is a globular protein which hydrolyzes sucrose in living organisms. Invertase widely distributed in the biosphere especially in baker's yeast and plants. Invertase enzyme has recently been used in various biotechnological applications such as beverage, confectionary, bakery, invert sugar, high fructose syrup, artificial honey, calf feed, food for honeybees and other applications. Thus, invertase is an interesting enzyme which is widely used in industrial processes, however, there are no comprehensive reviews published on invertase enzymes. PubMed, Scopus, and Google Scholar search engines were searched from January 2013 to November 2019. Based on inclusion and exclusion criteria 200 relevant experimental studies with moderate bias were selected. This paper provides comprehensive information on the invertase and their types, sources, production methods, physical and chemical properties, structures, separation and purification methods, and their applications.

31 citations

Journal ArticleDOI
TL;DR: Overall results demonstrated the feasibility of aqueous two phase extraction for the isolation and purification of invertase without the need of multiple steps.

29 citations

References
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Journal Article
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.

289,852 citations


"Extraction, partial purification an..." refers methods in this paper

  • ...Protein concentration was determined colorimetrically by Lowry method [18] using bovine serum albumin as a standard....

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Journal ArticleDOI
15 Aug 1970-Nature
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Abstract: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four major components of the head are cleaved during the process of assembly, apparently after the precursor proteins have assembled into some large intermediate structure.

232,912 citations


"Extraction, partial purification an..." refers methods in this paper

  • ...The peak with the invertase activity was checked for purity using 10% SDS-PAGE (75 μl protin was loaded) according to the method of Laemmli [15]....

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  • ...10% SDS-PAGE as previously described [15] using the same molecular weight markers used in gelfi ltration....

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Journal Article
01 Jan 1970-Nature
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Abstract: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four major components of the head are cleaved during the process of assembly, apparently after the precursor proteins have assembled into some large intermediate structure.

203,017 citations

Journal ArticleDOI
TL;DR: In this article, a method was developed to determine submicro amounts of sugars and related substances using a phenol-sulfuric acid reaction, which is useful for the determination of the composition of polysaccharides and their methyl derivatives.
Abstract: Simple sugars, oligosaccharides, polysaccharides, and their derivatives, including the methyl ethers with free or potentially free reducing groups, give an orangeyellow color w-hen treated with phenol and concentrated sulfuric acid. The reaction is sensitive and the color is stable. By use of this phenol-sulfuric acid reaction, a method has been developed to determine submicro amounts of sugars and related substances. In conjunction with paper partition chromatography the method is useful for the determination of the composition of polysaccharides and their methyl derivatives.

45,381 citations


"Extraction, partial purification an..." refers background in this paper

  • ...Phenol, in the presence of sulfuric acid, can be used for quantitative calorimetric micro determination of sugars and their methyl derivatives, oligosaccharide and polysaccharides as described by Dubois [21]....

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  • ...[21] reported 4....

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