{"title":"β-Fructofuranosidase from grape berries II. Solubilization of a bound fraction","authors":"Wilfred Niels Arnold","doi":"10.1016/0926-6593(66)90148-2","DOIUrl":null,"url":null,"abstract":"<div><p></p><ul><li><span>1.</span><span><p>1. A bound fraction of grabe <span><math><mtext>β-</mtext><mtext>fructofuranosidase</mtext></math></span> (EC 3.2.1.26) was solubilized by treatment with 0.2 M borate buffer (pH 8.5), whereas several other aqueous salts were without effect.</p></span></li><li><span>2.</span><span><p>2. In the system described, solubilization was influenced by boric acid concentration and pH but not by presentation time. Solubilization was essentially irreversible.</p></span></li><li><span>3.</span><span><p>3. Solubilization was maximal at pH 8.5; under more alkaline conditions, enzyme inactivation occurred. The decrease in amount of bound enzyme followed an inverse trend.</p></span></li><li><span>4.</span><span><p>4. Borate solubilization gave a total yield of <span><math><mtext>β-</mtext><mtext>fructofuranosidase</mtext></math></span> about 190% based on the starting material. This, together with the well-known interaction of borate with carbohydrates, suggested that a masking polysaccharide was operative in the starting material.</p></span></li><li><span>5.</span><span><p>5. The solubilized and soluble enzyme were partially purified and characterized. In both cases, only unsubstituted β-fructofuranosides were hydrolyzed. The Michaelis constants for sucrose were not significantly different.</p></span></li><li><span>6.</span><span><p>6. The dangers inherent in comparing the relative amounts of enzymes from different tissues were underlined by this experience.</p></span></li></ul></div>","PeriodicalId":100160,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1966-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0926-6593(66)90148-2","citationCount":"22","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0926659366901482","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 22
Abstract
1.
1. A bound fraction of grabe (EC 3.2.1.26) was solubilized by treatment with 0.2 M borate buffer (pH 8.5), whereas several other aqueous salts were without effect.
2.
2. In the system described, solubilization was influenced by boric acid concentration and pH but not by presentation time. Solubilization was essentially irreversible.
3.
3. Solubilization was maximal at pH 8.5; under more alkaline conditions, enzyme inactivation occurred. The decrease in amount of bound enzyme followed an inverse trend.
4.
4. Borate solubilization gave a total yield of about 190% based on the starting material. This, together with the well-known interaction of borate with carbohydrates, suggested that a masking polysaccharide was operative in the starting material.
5.
5. The solubilized and soluble enzyme were partially purified and characterized. In both cases, only unsubstituted β-fructofuranosides were hydrolyzed. The Michaelis constants for sucrose were not significantly different.
6.
6. The dangers inherent in comparing the relative amounts of enzymes from different tissues were underlined by this experience.