Purification and Properties of Yeast Invertase<sup>*</sup>

Np, Neumann; Jo, Lampen

doi:10.1021/bi00854a015

Cited by 239 publications

(92 citation statements)

References 36 publications

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“…The purified enzyme is probably a glycoprotein based on its ability to bind to the lectin Con-A. This is consistent with other studies (4,18,23) where it has been shown that invertase is a glycoprotein. The precise role of the carbohydrate moiety of the enzyme is not well understood since it is not necessary for enzymic activity or stability (4).…”

Section: Resultssupporting

confidence: 91%

Wheat Invertases

Krishnan¹,

Blanchette²,

Okita³

1985

Plant Physiol.

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Wheat coleoptiles have two distinct invertases, a soluble and a cell wall-bound form as indicated by results from cytochemical and biochemical studies. These enzyme activities differ in their pH optima, chromatographic behavior on diethyaminoethyl cellulose, kinetic properties, thermal stability, and response to light treatment. The soluble invertase was purified to near homogeneity by diethylaminoethyl-cellulose, concanavalin-A Sepharose, and Sephacryl S-300 chromatography. MATERIALS AND METHODS Buffer Solutions. The buffer solutions used were as follows: A, 20 mm sodium phosphate (pH 6.5), 1 mm EDTA, 1 mM DTT, 0.1 mm phenylmethylsulfonyl fluoride, and 1 mM benzamidine; B, 10 mm sodium phosphate (pH 6.5), 1 mM EDTA, and 0.1 mM DTT; C, 100 mm sodium phosphate (pH 6.5), 1 mM MnCI2, 1 mM CaCl2, 1 mM MgCl2, 1 mM DTT, and 0.5 M NaCl; D, 50 mm sodium phosphate (pH 6.5), and 1 mM DTT.Plant Material. Seeds of Triticum aestivum L. cv Yecovo Rojo were sown on two layers of filter paper moistened with distilled H20 on Petri plates. One set was grown under continuous light and the other placed in total darkness at room temperature. The coleoptiles and roots were harvested after 4 d and analyzed for invertase activity as discussed below.Invertase Extraction. Five g of tissue were homogenized with a mortar and pestle containing 30 ml of buffer A and then centrifuged at 3000g for 20 min. The resulting pellet was washed by repeated cycles of suspension in 30 ml of buffer A and centrifugation as described above. All supernatant fluids were combined and centrifuged at 31,000g for 20 min, and the clear supernatant fluid was designated as the soluble invertase fraction. The final pellet was resuspended in 30 ml of buffer A containing 1 M NaCl and shaken overnight at 4C. The resuspended pellet was then centrifuged at 31,000g for 20 min, and the resulting supernatant fluid was designated as the cell wall enzyme fraction. Both the soluble and cell wall enzyme fractions were dialyzed separately against buffer B to remove endogenous reducing sugars.Invertase Assay. Appropriate amounts of an enzyme preparation were added to a test tube containing 2 ml of20 mm sucrose in 50 mm sodium acetate buffer, pH 5.5, and incubated at 37C for 30 min. The reaction was terminated by adding 2 ml of Somagi reagent (17)

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Section: Resultssupporting

confidence: 91%

Wheat Invertases

Krishnan¹,

Blanchette²,

Okita³

1985

Plant Physiol.

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“…form secreted into the periplasmic space and an apparently non-glycosylated form retained within the cell (Neumann & Lampen, 1967;Lampen, 1968). The glycosylated form has been used as a probe for the process of glycoprotein biosynthesis and secretion (Novick et al, 1981;Trimble et al, 1983;Esmon et al, 1987).…”

Section: Introductionmentioning

confidence: 99%

Regulation of invertase in Aspergillus nidulans: effect of different carbon sources

Vainstein¹,

Peberdy

1991

Microbiology

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Aspergillus nidulans produces an extracellular P-D-fructofuranoside fructohydrolase (invertase) when grown on a medium containing the P-fructofuranosides sucrose or raffinose, indicating that synthesis is subject to induction by the substrate. On a growth medium containing sucrose, production was maximal at 15 h in cultures incubated at 28 Co. After this time the level of detectable invertase in the cultures declined. A proportion of the enzyme was secreted during the linear growth phase of the fungus. Various sugars were investigated for induction of invertase, but only the two P-fructofuranosides induced high production levels; with the other sugars, the enzyme was produced only at a low constitutive level. Mycelium grown under repressive conditions (1% glucose), rapidly produced invertase when transferred to sucrose-containing medium. After 80 min the invertase level in these cultures was 26-fold higher than the constitutive level. The repressive effect of other sugars, e.g. glucose and xylose, on invertase production was also demonstrated in this experimental system.

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“…The same production medium used in secondary screening was used for optimization studies. In all the experiments the flasks were inoculated with 2 ml of inoculum prepared from the slant cultures [8].…”

Section: Optimization Studiesmentioning

confidence: 99%

Production of Invertase by Aspergillus niger Under Solid State Fermentation Using Orange Fruit Peel as Substrate

Raju¹,

King²,

Jetti³

2016

Adv Crop Sci Tech

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The Aspergillus niger was tested for its ability to produce invertase on natural agricultural residues. Orange peel was selected as substrate due to its high peak enzyme activity, the maximum invertase activity was obtained in preliminary studies by incubating flasks containing orange peel as substrate supplemented with sucrose (1.0% w/v), yeast extract (1% w/v), and 4 ml of A. niger conidial suspension for 96 hours at 30°C.

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Purification and Properties of Yeast Invertase^*

Cited by 239 publications

References 36 publications

Wheat Invertases

Wheat Invertases

Regulation of invertase in Aspergillus nidulans: effect of different carbon sources

Production of Invertase by Aspergillus niger Under Solid State Fermentation Using Orange Fruit Peel as Substrate

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Purification and Properties of Yeast Invertase*

Cited by 239 publications

References 36 publications

Wheat Invertases

Wheat Invertases

Regulation of invertase in Aspergillus nidulans: effect of different carbon sources

Production of Invertase by Aspergillus niger Under Solid State Fermentation Using Orange Fruit Peel as Substrate

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Purification and Properties of Yeast Invertase^*