• Journal of Ginseng Research
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• v.14 no.1
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• pp.14-20
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• 1990

In An invertase (EC 3.2.1.26) was extracted from Korean giseng (Panax ginseng C.A. Meyer) with distilled tvater The ginseng invertase was purified about 62.6 folds purified by procedures including ammonium sulfate fractionation , DEAE-cellulofine chromatography and gelfiltrations through Sephadex G-75 and the recovery of enzyme activity was 11.1%. The homogeneity of the purified enzyme was probed by polyacrylamide gel disc electrophoresis. The purifled enzyme was divided into two different subunits by treating with a mixture of SDS and 2-mercautoethanol, and the molecular weight of the large subunit was estimatedtobe 116,000 and that of the small one to be 14,000. The optimal VH and temperature of the enzyme were pH 6 and 45$^{\circ}C$, respectively. The enzyme hydrolyzed specifically the hydrolyzation of the -fructofuranosides such as sucrose, raffinose and inulin. The Km values of the enzyme for sucrose and raffinose were determined to be 0.85 and 0.6 mM, respectively.

• Journal of the Korean Society of Food Culture
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• v.5 no.2
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• pp.269-274
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• 1990

This study was conducted to determine invertase activity in persimmon during the drying process and characterize the purified enzyme. As drying proceeded, invertase activity increased until 10 days and decreased gradually afterwards. Invertase in persimmon fruit was extracted with 250 mM potassium phosphate sulfate buffer at pH 7.4. The enzyme was purified by means of ammonium sulfate fractionation, column chromatography on DEAE-cellulose and gel filtration on Sephadex G-200 column. The optimal temperature of enzyme was $40^{\circ}C$ and optimal pH was 5.0 and 6.0 for sucrose and raffinose, respectively. The enzyme was stable up to $50^{\circ}C$ and pH 3-6. The Km value of the enzyme, with sucrose as a substrate, was 2.5mM. Electrophoretic pattern of purified enzyme solution showed a single band.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.368-375
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• 1990

Rhodoto& ghtbth~ K-24 was found to produce external invertase in addition to internal and cell wall bound invertase. External invertase was purified to an electrophoretically homogeneous state and partitally characterized and was compared with internal and cell wall bound invertase of which procedures for purification and characterization were reported previously. The enzyme was purified by ethanol precipitation, column chromatographies on DEAE-Sephadex A-50 and SP-Sephadex C-50, and gel filtration on Sephadex G-100. The molecular weight and subunit molecular weight of external invertasGwere estimated to be 220,000 and 100,000, respectively. The isoelectric point of the enzyme was about pH 6.0. The optimum pH and temperature for enzyme activity were pH 4.0 and $60^{\circ}C$, respectively. The enzyme remained stable at the wide range, from pH 3.0 to 11.0 and stable up to $40^{\circ}C$, but was inactivated at temperatures above that. $HgC_12, AgN0_3, MnS0_4$, SDS and p-CMB inhibited the enzyme activity. The $K_m$ value of the enzyme for sucrose was $1.0\times 10^{-2}$M. From these results, the three isozymes from Rh. glutinis K-24 seem to have the similar enzymatic properties, but to differ in molecular and subunit weights.

• Journal of Ginseng Research
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• v.14 no.1
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• pp.21-26
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• 1990

The chemical composition and stabilities of the purified ginseng invertase were investigated. The purified enzyme was found to be a glycoprotein composed of 80.2% protein and 19.7% total sugar. The protein component of the enzyme was composed of acidic amino acid (9.3%), basic amino acid (48.9%), nonpolar amino acid (21.4%), polar amino acid (20.4%) and 6.1% S-containing amino acid. It showed especially high contents of histidine and serine. The enzyme was inactivated almost completely by the treatment with some proteases (papain, pepsin. trypsin, pancreatin and microbial alkaline pretense) and protein denatllrants (8M urea and 6M guanidine-HC1), bolt not with glyrosidase (${\alpha}$-amylase, ${\beta}$-amylase. glcoamylese and cellullase). btonosaccharides sllch as glilrose, fructose, galactose and mannose did not exert any influence on the enzyme activity. The activity of the enzyme was inhibited by Ag+, Mn2+, Hg2+, Zn2+ and Al3+, whereas Ca2+, Mg2+, Ba2+ and Fe3+ gave rather activating effects on the enzyme activity. The enzyme was relatively stable in the VH range of VH 6 and 8, and at the temperatures below 35$^{\circ}C$.

• Preventive Nutrition and Food Science
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• v.2 no.3
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• pp.250-254
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• 1997

The internal invertase of Rhodosporidium toruloids mating type a cells was purified to a single band on SDS-PAGE from cell-free extract by acid precipitation, ion exchange chromatogaphy andgel filtration. The determined molecular weight of he purified enzyme was about 95,000 by gel filtration and 100,000 daltons on SDS-polyacryamide gel electrophoresis. This enzyme didn't show any activity change by several metal ions except 15.4% decrease by {TEX}$Mn^{2+}${/TEX} and was strongly inhibited by 2-mercaptoethanol and SDS. The invertase maintained its activity at high level until 70℃, but inactivated at 80℃ almost completely. The optimal temperature and pH of the enzyme were about 60℃ and pH 5.0, respectively. The stable pH range of invertase was narrow from pH 3.0 to 6.0. The Km value and isoelectric point of enzyme were 3.4×{TEX}$10^{3}${/TEX} M, pH 4.4, respectively.

• The Korean Journal of Food And Nutrition
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• v.2 no.2
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• pp.8-13
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• 1989

A strain of Saccharomyces cerevisiae BY-366 was found to produce a strong sucrose-hydrolyzing enzyme Using this strain, the optimal culture conditions for the production of invertase were investigated. The results are as follows : 1. For enzyme production, optimal temperature, initial pH and critical concentrations of sucrose and raffinose were 3$0^{\circ}C$, 5.0 and 3.0%, respectively. 2. Enzyme production was reached maximum by organic nitrogen source, 0.3% yeast extract plus 0.5% bactopeptone. 3. It was appeared the presence of 0.1 M Mn2+ and Fe2+ ion was essential factors, on the other hand, 0.1 M Ag+ and Hg2+ ion almost block in yeast growth and enzyme production. 4. Invertase productivity was reached maximum within 3 days on stationary culture with medium-composed of sucrose 3%, bactopeptone 0.5%, yeast extract 0.3%, KEHPO. 0,1%, MgSO4.7H2O 0.05%.

• KSBB Journal
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• v.25 no.1
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• pp.79-84
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• 2010

In the present study, we isolated a new bacterial strain producing invertase (EC 3.2.1.26) and determined optimized culture condition in flask culture. The strain was identified as Bacilus flexus determined by the 16S rDNA sequencing method. The invertase was produced only in the sucrose medium as the sole carbon source. Potassium nitrate was an adequate nitrogen source for enzyme production, whereas meat peptone showed the highest bacterial growth. Enzyme production was increased about 2-fold when $MgSO_4\cdot7H_2O$ was supplemented to the growth media. The optimum temperature was found to be $30^{\circ}C$ for both enzyme production and bacterial growth. Invertase exhibited pH optima in the range 5.0-6.0 and have a temperature optimum at $40^{\circ}C$, similarly to other invertases found from different microbial sources. Several mineral ions (K and Fe) stimulated the invertase activity, whereas some bioelements (Ag, Mg, and Mn) inhibited enzyme activity. Under the optimized culture condition, the maximum enzyme production (over 250 units/mL) was achieved at 20 h. To the best of our knowledge, this is the first time to report on invertase production by Bacilus flexus.

• Journal of Environmental Science International
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• v.4 no.4
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• pp.21-21
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• 1995

The effects of plant hormones (NAA, $GA_3$ and BA) and light qualities (white, red, green and blue light) on the changes of reducing sugar contents and invertase isozyme activities in leaves of maize (Zea mars L.) and mung bean (Phseolw radiatus L.) seedlings were investigated. NAA accelerated the increase of reducing sugar contents and invertase isozyme activities, on the contrary, $GA_3$ had little effect in the accumulation of reducing sugar and in the increase of enzyme activities from the leaves of maize and mung bean seedlings. On the other hand, BA accelerated an increase in the activities of the invertase isozyme from the leaves of mung bean seedlings whereas it had little effect in the increase of the enzyme activities from those of maize seedlings. The accumulation of reducing sugar in leaves of both seedlings was promoted by red light irradiation compared to white light irradiation, while the activities of the enzyme were little affected by various light Qualities. In the simultaneous applications of plant hormone and light quality, NAA with white light was very effective in the increase of reducing sugar contents and the enzyme activities from the leaves of mung bean seedlings, whereas NAA application with blue light showed a prominent enhancement in the reducing sugar contents and the enzyme activities from those of maize seedlings. These results suggest that plant hormone, particularly NAA, may be a more important factor than various light Qualities in the stimulation of invertase activity.

• Journal of Environmental Science International
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• v.4 no.4
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• pp.323-333
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• 1995

The effects of plant hormones (NAA, $GA_3$ and BA) and light qualities (white, red, green and blue light) on the changes of reducing sugar contents and invertase isozyme activities in leaves of maize (Zea mars L.) and mung bean (Phseolw radiatus L.) seedlings were investigated. NAA accelerated the increase of reducing sugar contents and invertase isozyme activities, on the contrary, $GA_3$ had little effect in the accumulation of reducing sugar and in the increase of enzyme activities from the leaves of maize and mung bean seedlings. On the other hand, BA accelerated an increase in the activities of the invertase isozyme from the leaves of mung bean seedlings whereas it had little effect in the increase of the enzyme activities from those of maize seedlings. The accumulation of reducing sugar in leaves of both seedlings was promoted by red light irradiation compared to white light irradiation, while the activities of the enzyme were little affected by various light Qualities. In the simultaneous applications of plant hormone and light quality, NAA with white light was very effective in the increase of reducing sugar contents and the enzyme activities from the leaves of mung bean seedlings, whereas NAA application with blue light showed a prominent enhancement in the reducing sugar contents and the enzyme activities from those of maize seedlings. These results suggest that plant hormone, particularly NAA, may be a more important factor than various light Qualities in the stimulation of invertase activity.

• Journal of Plant Biology
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• v.38 no.3
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• pp.251-258
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• 1995

The soluble acid invertase ($\beta$-D-fructofuranoside fructohydrolase, EC 3.2.1.26) was isolated and characterized from the hypocotyls of mung bean (Phaseolus radiatus L.). The enzyme was purified to apparent homogeneity by consecutive step using diethylaminoethyl (DEAE)-cellulose anion exchange, Concanavalin (Con) A affinity and Sephacryl S-300 chromatography. The overall purification was about 148-fold with a yield of about 15%. The finally purified enzyme exhibited a specific activity of about 139 $\mu$mol of glucose produced mg-1 protein min-1 at pH 5.0 and appeared to be a single protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing PAGE. The enzyme had the native molecular weight of 70 kD and subunit molecular weight of 70 kD as estimated by Sephadex G-200 chromatography and SDS-PAGE, respectively, suggesting that the enzyme was composed of a monomeric protein. On the other hand, the enzyme appeared to be a glycoprotein containing N-linked high mannose oligosaccharide chain on the basis of its ability to bind to the immobilized C on A. The enzyme had a Km for sucrose of 1.8 mM at pH 5.0 and maximum activity around pH 5.0. The enzyme showed highest enzyme activity with sucrose as substrate, but the activity was slightly measured with raffinose and cellobise. No activity was measured with maltose and lactose. These results indicate the soluble acid invertase is a $\beta$-fructofuranosidase.