Adsorption and chemisorption of galactose oxidase on silica surface
Experience accumulated over a number of years in developing of methods of immobilization of galactose oxidase from Fusarium graminearum on parent and modified silica matrices is analyzed. Sturdy adsorption of galactose oxidase on silica surface was observed, such heterogeneous specimens possessed by...
Saved in:
| Date: | 2002 |
|---|---|
| Main Authors: | , , |
| Format: | Article |
| Language: | English |
| Published: |
Chuiko Institute of Surface Chemistry National Academy of Sciences of Ukraine
2002
|
| Online Access: | https://surfacezbir.com.ua/index.php/surface/article/view/93 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Journal Title: | Surface |
| Download file: |  |
Institution
Surface| _version_ | 1869291184273227776 |
|---|---|
| author | Kondakova, L. V. Yanishpolskii, V. V. Tertykh, V. A. |
| author_facet | Kondakova, L. V. Yanishpolskii, V. V. Tertykh, V. A. |
| author_institution_txt_mv | [
{
"author": "L. V. Kondakova",
"institution": "Інститут хімії поверхні НАН України"
},
{
"author": "V. V. Yanishpolskii",
"institution": "Інститут хімії поверхні НАН України"
},
{
"author": "V. A. Tertykh",
"institution": "Інститут хімії поверхні НАН України"
}
] |
| author_sort | Kondakova, L. V. |
| baseUrl_str | |
| collection | OJS |
| datestamp_date | 2018-11-27T09:42:19Z |
| description | Experience accumulated over a number of years in developing of methods of immobilization of galactose oxidase from Fusarium graminearum on parent and modified silica matrices is analyzed. Sturdy adsorption of galactose oxidase on silica surface was observed, such heterogeneous specimens possessed by enhanced biocatalyst stability and activity as compared with enzyme solutions. Covalent immobilization of galactose oxidase was carried out on the amine-containing silicas activated by 2,4-tolylene diisocyanate and cyanuric chloride. It was also shown that in the presence of the substrate (galactose) enzyme chemisorption takes place on the surface on amine-containing silica matrices. Immobilized preparations were successfully applied for analytical determination of galactose-containing carbohydrates (galactose, lactose, raffinose) in complex mixtures. |
| first_indexed | 2025-07-22T19:30:12Z |
| format | Article |
| fulltext |
150
ADSORPTION AND CHEMISORPTION OF GALACTOSE
OXIDASE ON SILICA SURFACE
L.V. Kondakova, V.V. Yanishpolskii, and V.A. Tertykh
Institute of Surface Chemistry, National Academy of Sciences
Gen. Naumov Str. 17, 03680 Kyiv-164, UKRAINE
Abstract
Experience accumulated over a number of years in developing of methods of immobilization
of galactose oxidase from Fusarium graminearum on parent and modified silica matrices is analyzed.
Sturdy adsorption of galactose oxidase on silica surface was observed, such heterogeneous
specimens possessed by enhanced biocatalyst stability and activity as compared with enzyme
solutions. Covalent immobilization of galactose oxidase was carried out on the amine-containing
silicas activated by 2,4-tolylene diisocyanate and cyanuric chloride. It was also shown that in the
presence of the substrate (galactose) enzyme chemisorption takes place on the surface on amine-
containing silica matrices. Immobilized preparations were successfully applied for analytical
determination of galactose-containing carbohydrates (galactose, lactose, raffinose) in complex
mixtures.
Introduction
Galactose oxidase (D-galactose: EC 1139) came to the attention in the connection with possibility
to use for analytical determination of D-galactose and galactose-containing sugars. In this case oligo-
and polysugars, which contain D-galactose, display a high affinity to the enzyme, and they are
oxidized fast by galactose oxidase (GalO) [1-4]. An ability of GalO to stereospecific oxidation of the
tribasic alcohols may be used for enzymatic synthesis of some optically pure isomers. Many of the
difficulties, connected with use of the soluble GalO in analysis and synthesis, can be successfully
solved by application of immobilized enzyme preparations. As known, in the majority of cases a
bound enzyme becomes more stable, the immobilized biocatalyst may be easily separated from the
reaction products and used repeatedly. At the same time the data, devoted to methods of obtaining
and studies of properties of the immobilized GalO preparations, are limited in number [5-12]. In the
present work, experience, which has been accumulated over a number of years in the field of using of
active and stable preparations of GalO for analytical purposes and in organic synthesis, is discussed.
Such preparations can be obtained both as a result of the enzyme adsorption on the silica and by
covalent anchoring on the modified surface.
Experimental
Silochrom S-80, macroporous silica (prepared from fumed silica, the Stavropol plant of pure
chemical reagents and luminophores) with specific surface area 80 m2/g, the total pore volume 1.2
cm3/g, and the particle size 0.25-0.50 mm was applied as a support for the GalO adsorption and
immobilization. GalO preparations were obtained from Fusarium graminearum IMV-1060. They
have been used in solutions and in the form of liophilic-dried powder [13]. For the determination of
activity of soluble and immobilized GalO preparations D-galactose (purified grade, may contain 1-
2% glucose) was used as a substrate. D-galactose solutions were prepared in the form of 100 mM
solution in 0.05 M phosphate buffer (pH=7.0), containing 100 mM potassium ferricyanide and 1M
Na-EDTA [11].
151
The adsorption of galactose oxidase was carried out in the following way: the previously
prepared buffer enzyme solution was mixed with a weighted specimen of the carrier and
incubated at 25oC for 2 h. Then, for removing of an unbound enzyme the carrier was washed
by the buffer solution [14]. In this case about 10 ml of the buffer solution on 1g of a carrier
was usually applied.
Covalent attachment of GalO was carried out on the modified amine-containing silicas,
activated by cyanuric chloride (a) and 2,4-tolylene diisocyanate (b) [11, 15, 16]. In these cases
enzyme anchorage are achieved by the nitrogen-carbon (a) and urine bonds (b) formation in
accordance with the following schemes:
N
N
N
SiCH2CH2CH2NH
NH Enzyme
Cl
(a)
SiCH2CH2CH2NHC(O)NH CH3
NHC(O)NH Enzyme (b)
Syntheses of the activated silica carriers were executed by methods described in our
previous works [15, 16]. The immobilization process was carried out at room temperature by
proceeding as follows: to 100 mg of the appropriate activated support was added the enzyme
solution in 0.05 M phosphate buffer (pH=7.0) and mixture was stirred for 2 h. Thereafter the
carrier with immobilized biocatalyst was washed by the buffer solution.
As is shown in [17] and this work, when a solution contains galactose oxidase and its
substrate (galactose), a peculiar process of autoimmobilization of the enzyme on the
amine-containing silica (aminoorganosilica) surface takes place. In this situation, under the
action of a biocatalyst galactose is transformed into 2,4-galactohexodialdose that serves as a
cross-linking agent and provides bonding of the enzyme with the amine groups of the modified
matrix surface with formation of azomethine bonds. Thus, in this case the immobilization may
be carried out through the mixing of the buffer enzyme solution, galactose and
aminoorganosilica in the various ratios. Usually 50 ml of the galactose solution per 1g of a
carrier is used. After incubation of the mixture for 2 h at room temperature, for removing of an
unbound enzyme the immobilized preparations were washed by the buffer. In the control
experiments the GalO adsorption was performed onto the same aminoorganosilica carrier in
the absence of the substrate.
The activity of the soluble GalO was evaluated per a rate of oxygen consumption under the
certain conditions employing the Clark oxygen electrode. The activity of the immobilized GalO
was determined in the flow-type reactor. Measurements were done both in the impulsive and
continuous regimes of a substrate flowing through a column with the immobilized biocatalyst.
In the former case oxygen consumption is registered as a peak and in the latter case as a
stationary O2 concentration at the output of the reactor.
Results and discussion
GalO adsorption on the silica surface. The very firm GalO adsorption has been observed
on the initial silica surface. We could not completely remove enzyme from the carrier surface
by buffer solutions with the various pH (4.5, 5.5, 8.2) and molarity. This is due to the fact that
152
in water the silica particles go negative charge and the enzyme macromolecules have clearly
defined positive charge [18]. It should be particularly emphasized that in the most of cases at
various enzyme/carrier ratios, taking for immobilization, it has been possible to obtain the
heterogeneous specimens with an enhanced enzyme activity (in the some of cases enough
essential) as compared with the enzyme activity in the starting solution (Table 1). It should be
also noted that the pH-optimum of the soluble GalO was found at pH=7.0 and the deviations
from this value reduce the enzyme activity. It is significant that preparations of adsorbed GalO
possess sufficiently more high stability as compared with enzyme solutions. The activities of
immobilized specimens are still retained for several months of storage. It is suggested [6] that
enzyme attachment to surface of the support, which is unaffected by temperature or pH
alteration, provides a biocatalyst by the protective frame and preserves enzyme from the
conformational changes. It is quite possible that in the case of GalO adsorption on the silica
surface high conformational stability of the biocatalyst and thus its high activity are available.
Similar, but more significant effect was observed [19] in time of an incubation of the enzyme
by phosphate “bridges”, which have also stabilized the conformation of the protein molecules.
Table 1. Dependence of activity of galactose oxidase adsorbed by silica on
concentration of soluble enzyme in initial solution (optimum pH=7.0).
Example
Enzyme activity in initial solution,
Int. units*/ml
Enzyme/Silica
ratio taken for
adsorption,
Int.units/g
Activity of
adsorbed
preparation,
Int. units/g
1 0.13 1.3 28.6
2 0.01 0.1 2.4
3 0.25 2.5 13.0
4 0.002 0.02 0.1
5 0.27 2.7 2.6
*1 international unit of activity is equal to enzyme quantity, which catalyzes an
oxidation
of 1 mmol of the substrate for 1 min.
As follows from obtained data, of great importance for obtaining of the active preparations
is an enzyme/carrier ratio taken for adsorption. This can best be done in the buffer solutions
where the enzyme/carrier ratio is equal to 1.3 units/g (international units of an enzyme activity
per 1 g of a carrier). The deviation from this value causes decrease in the specimen activity.
Nevertheless, in these cases relatively active heterogeneous preparations were obtained. At
increasing of an enzyme/carrier ratio up to 2.7 (Table 1, sample 5) a growth of activity of the
enzyme was not detected at all. The observed trend can result from an enzyme inactivation
because of protein/protein interactions at high surface loadings. Less active specimens were
prepared at low surface loadings. It is conceivable that the observed results may be explained
by strong electrostatic interactions of adsorbed enzyme molecules with the surface sites. In a
case of such interaction the considerable conformational changes in the GalO molecule may
occur resulting in a lowering of the immobilised enzyme activity.
Variations in the pH values and buffer type (pH=4.5, 5.8, 6.1, 8.2, and 9.1; Na-acetate and
Tris-HCl buffer) have a small effect on a degree of the enzyme activation at sorption on the
silica surface. However, the enzyme stability is pH-dependent, and it may be connected with
distribution of protons and presence of the ionized groups on a matrix surface. Therefore the
153
obtained preparations of adsorbed GalO are usually characterized by more low stability as
compared with the specimens prepared at the optimal pH-value.
Covalent GalO immobilization. As known, stable preparations of immobilized enzymes,
which are not removed from the carrier surface at changing of pH and ionic strength of a
solution, may be obtained at covalent binding. In our previous works [10, 11] we proposed
methods for GalO chemical binding to surface of the silica matrix. For this purpose
amine-containing silicas, activated by cyanuric chloride and 2,4-tolylene diisocyanate, were
applied. The use of these activating reagents, whose groups of the same composition differ
substantially in their chemical activity, permits one to carry out a process of bonding in two
steps. First, a reaction is brought about for amine groups of the modified silica matrix, and then
functional groups of an enzyme subjected to immobilization enter into the reaction. In
particular, isocyanate groups of 2,4-tolylene diisocyanate bound to benzene rings in position 4
are more active than NCO group in position 2. Therefore, when amine-containing silica comes
into contact with solutions of 2,4-tolylene diisocyanate in carbon tetrachloride, it is isocyanate
groups in position 4 that predominantly enter into the surface reaction. The modified silica
matrices activated in this way contain grafted isocyanate groups in position 2 of benzene rings.
Then the grafted isocyanate groups interact with amine groups of enzyme macromolecule. In a
six-member triazine ring of cyanuric chloride in position 2,4,6 there are ºCCl groups with a
highly labile chlorine atom. Interaction between one of these bonds (C-Cl) of the activating
agent and a grafted aminopropyl group can result in formation of chemical compounds
attached to the surface by hydrolytically stable C-NH-C bonds. It is significant that if one of
the ºCCl groups of cyanuric chloride molecule enters into the reaction, the activity of the rest
of the C-Cl bonds in the surface chemical compounds formed slightly decreases. At the same
time, the mobility of chlorine atoms in such C-Cl bonds remained sufficiently high to allow
bonding of enzyme molecules under mild conditions.
The data represented in the Table 2 show that in a case of using for immobilization of the
GalO solutions with the activity 12-80 units the obtained immobilized preparations preserve
75-100% of initial activity. It should be noted that a growth of the enzyme activity at chemical
binding with surface do not occur, as it is in a case of the GalO adsorption on silica surface. If
the enzyme solutions, having more high activity (115-140 units), were used for covalent
attachment, the preparations obtained conserved only 54-59% of an initial activity. It is
conceivable that at more high concentrations of protein the role of protein-protein interactions
increase resulting in a loss of activity. It is significant that the influence of activation method of
amine-containing silica surface (by cyanuric chloride or 2,4-tolylene diisocyanate) on the
activity of immobilized preparations obtained be not detected (Table 2).
Immobilization by product of the enzymatic reaction. As known, GalO catalyzes an
oxidation of D-galactose in the C-6 position by molecular oxygen. The enzymatic reaction
product is dialdehyde - 2,4-galactohexodialdose. We know from experience that dialdehydes
(glutaraldehyde, gossypol) are effective cross-linking reagents, which provide covalent binding
of enzymes and other biologically active compounds to the surface of the amine-containing
silicas. It has been found that 2,4-galactohexodialdose at the neutral pH-values can react with
amine groups both the enzyme and matrix surface. From the data, represented in the Table 3,
one can see that an activity of immobilized preparations depends on the ratio of the taken
components. In the result of interaction of amine-containing carrier, enzyme and substrate the
immobilized enzyme specimens were obtained. These preparations preserved their activity for
2 months in the condition of flow-injection analyzer (passing through 50 L of a buffer and
1000-fold volume of a substrate). The stability of the obtained preparations is no different from
stability enzyme, immobilized by covalent binding to modified silica surfaces activated by
cyanuric chloride or 2,4-tolylene diisocyanate. Without addition of the substrate (galactose) we
154
have observed the enzyme desorption after washing by two volumes of 0.05 M phosphate
buffer. It is obvious that positive charge of amine-containing silica surface
(ºSiCH2CH2CH2NH3
+) is not favoured to binding of the GalO molecules also having positive
charge [18].
Table 2. Dependence of activity of galactose oxidase immobilized on amine-containing
silicas activated by cyanuric chloride (AsilCC-sample) or 2,4-tolylene
diisocyanate (AsilTDI-sample) on activity of enzyme in initial solution.
Enzyme AsilCC-sample AsilTDI-sample
N/N activity in
initial solution,
Int. units
Activity of
immobilized
preparation,
Int. units
Degree of
activity
conservation, %
Activity of
immobilized
preparation,
Int. units
Degree of
activity
conservation,
%
1 23 21 90 20.5 89
2 115 69 60 68 59
3 12 12 100 12 100
4 140 75 54 76 54
5 80 59 74 60 75
Table 3. Immobilization of galactose oxidase on the surface of amine-containing silica
matrices in the presence of various amounts of substrate (galactose).
N/N
Galactose/Silica
ratio
Initial galactose
oxidase activity,
Int. units
Activity of
immobilized galactose
oxidase, Int. units
Degree of activity
conservation, %
1 0.36 46 9 20
2 2.88 46 45 98
3 0.18 46 4.5 10
4 3.24 46 45 98
5 1.44 46 38 83
6 0.90 23 20 87
7 0.90 115 89 77
8 0.90 140 89 64
9 0.90 12 12 100
10 0.90 80 75 93
11 - 23 1 4
12 - 69 4 6
13 - 115 5 4
155
Fig. 1. Oxygen expenditure at determination of
lactose (1), galactose (2), and raffinose
(3) by immobilized galactose oxidase on
addition of 3×10-6 M CuCl2 to 0.05 M
(pH=7.0) phosphate buffer.
Fig. 2. Dependence of O2 expenditure by
immobilized galactose oxidase on
glycerol concentration: 1 - without
activator; 2 - on addition of 100 mmol
K3[Fe(CN)6]; 3 - on addition of
Na-EDTA and CuCl2 (10-5 mol/L).
From the data in the Table 3, one can see that a high enzymatic activity of heterogeneous
preparations was observed at an addition 0.90-2.88 g of galactose per 1g of the carrier.
However further enhancement of galactose amount up to 3.24 g does not lead to a growth of
immobilized enzyme activity. In a similar manner a lowering of galactose amount to 0.18 and
0.36 g results in a lowering of activity of preparations immobilized on the amine-containing
silicas. At the designated galactose concentration (0.90 g/g) a high activities of immobilized
GalO preparations were detected at using of solutions containing 80-115 units of enzyme
activity.
As can be readily appreciated, the proposed method allows one to exclude some stages, in
particular a carrier surface activation, and obtain the active preparations of immobilized
enzyme. Besides, at using of activating agents it is not inconceivable that these reagents at the
interaction with enzyme can deactivate the active centres of biocatalyst [6]. It is hoped that this
is not the case when the product of enzymatic reaction is used for enzyme immobilization.
The obtained specimens of GalO, immobilized on the initial and modified silica matrices, are
stable and active preparations, which have considerable promise for the determination of
galactose-containing sugars and in the fine organic synthesis [20]. These preparations were
successfully used (Figs. 1, 2) for analytical determination of galactose-containing
carbohydrates in a complex mixture (the linear region for analysis: 0.1-20 mmol/L for
raffinose; 2-20 mmol/L for galactose; and 5-100 mmol/L for lactose).
Conclusions
Method of adsorption immobilization of galactose oxidase on the silica surface at the
optimal pH-value makes possible obtaining of active heterogeneous enzyme preparations. In
addition the resulting specimens possess many a time by more high activity as compared with
an enzyme activity in solution taken for adsorption. As this takes place, the carrier has no need
of a surface preliminary activation. Growth of an enzyme activity was not detected at covalent
156
binding of galactose oxidase on amine-containing silicas activated by cyanuric chloride or
2,4-tolylene diisocyanate. It must be indicated that method of activation has no noticeable
effect on activity and stability of immobilized preparations. It has been found that in the
presence of galactose covalent immobilization of galactose oxidase is rendered possible on
amine-containing silicas without a stage of activation of the carrier surface. It is suggested that
binding with surface amine groups is executed with the use of 2,4-galactohexodialdose,
forming at enzymatic oxidation of galactose. The adsorbed and chemically bound preparations
of galactose oxidase have considerable promise in analyses of its substrates and in some
organic syntheses.
Acknowledgement
The authors render thanks to T.T. Buglova and O.V. Skorobogat’ko from D.K. Zabolotny
Institute of Microbiology and Virology of National Academy of Sciences of Ukraine for
submitting of the samples of galactose oxidase from Fusarium graminearum IMV-1060.
References
1. Lukomskaya I.S., Ellanskaya I.A., and Gorodetskii V.K. Experimental Micology. – Kyiv:
Naukova Dumka. – 1968 (in Russian).
2. Gansedo J.M., Gansedo C., and Asensio C. Widespread occurrence of galactose oxidase
and glucose oxidase in fungi // Archives Biochem. Bioph.- 1967.-V.119, N1-3.- P.588-590.
3. Avigad G., Amoral D., Asensio C., and Horecker B.U. The galactose oxidase of Polyporus
circinatus // J. Biol. Chem.-1962.-V.237, N9.-P.2736-2743.
4. Klibanov A.M., Albert B.N., and Marletta M.A. Stereospecific oxidation of aliphatic
alcohols catalyzed by galactose oxidase // Biochem. Biophys. Res. Commun.- 1982.-
V.108, N2.- P.804-808.
5. Tertykh V.A. and Yanishpolskii V.V. Chemical aspects of enzyme immobilization on
inorganic matrices // Adsorption and Adsorbents. – 1980, N8. – P.3-27 (in Russian).
6. Triven M. Immobilized Enzymes. Moscow: Mir. –1983 (in Russian).
7. Hamilton G.A., De Jersey J., and Adolf P.K. Galactose oxidase: the complexities of a
simple enzyme. In: Oxidases and Related Redox Systems.- Baltimore- London: Univ., Park
Press., 1973.- P.103-119.
8. Pat. USA 4220503. - Jonson J., Stabilization of activated galactose oxidase enzyme, Appl.
28.04.78, Publ. 2.09.80.
9. Taylor R.J., Kmetec E., and Jonson J.M. Design, construction and application of a
galactose selective electrodes // Anal. Chem.- 1977.-V. 49, N6.-P.789-794.
10. Kondakova L.V., Yanishpolskii V.V., Tertykh V.A., Buglova T.T., and Koroleva O.V.
Some properties of galactose oxidase from Fusarium graminearum IMV-F-N1060
immobilized on aminoorganosilicas // Ukr. Biokhim. Zhurn. – 1984. – V.56, N4. –
P.394-398.
11. Kondakova L.V., Yanishpolskii V.V., Tertykh V.A., and Buglova T.T., Effect of
potassium ferricyanide and Cu(II) ions on properties of immobilized galactose oxidase //
Ukr. Biokhim. Zhurn. – 1988. – V.60, N6. – P.34-38.
12. Tertykh V.A. and Yanishpolskii V.V. Adsorption and chemisorption of enzymes and other
natural macromolecules on silicas // In: Adsorption on Silicas, Ed. by E. Papirer, Marcel
Dekker, New York, 2000, P.523-564.
13. Pat. USSR 889705. – Buglova T.T., Kirilenko T.S., and Zakharova I.Ya. Method of
preparation of galactose oxidase. – Byull. Izobret., 1981, N46.
157
14. Pat. USSR 1723124. – Kondakova L.V., Yanishpolskii V.V., Tertykh V.A., and
Buglova T.T., Method of galactose oxidase immobilization. – Byull. Izobret., 1992, N2.
15. Tertykh V.A., Yanishpolskii V.V., Chuiko A.A., Galich I.P., Tsyperovich A.S., and
Koval’chuk T.A. Immobilization of a-amilase on surface of highly dispersed silica //
Doklady AN UkrSSR, ser. B. – 1977. – N7. –P.654-657.
16. Yanishpolskii V.V., Tertykh V.A., and Lyubinskii G.V. Trypsin immobilization on
organosilica surfaces // Ukr. Biokhim. Zhurn. – 1979. – V.51, N4. – P.324-329.
17. Pat. USSR 1521775. – Kondakova L.V., Yanishpolskii V.V., Tertykh V.A., Buglova T.T.
and Skorobogat’ko O.V. Method of galactose oxidase immobilization. – Byull. Izobret.,
1989, N42.
18. Buglova T.T., Skorobogat’ko O.V., and Zakharova I.Ya. Characterization of purified
preparations of galactose oxidase from Fusarium graminearum IMV-F-N1060 //
Mikrobiol. Zhurn. – 1983. – V.45, N6. – P.47-52.
19. Rippa M., Signorini M., and Bellini T. The effect of inorganic phosphate on the stability of
some enzymes // Biochem. J.- 1982.-V.197, N3.-P.747-749.
20. Kondakova L.V., Yanishpolskii V.V., and Tertykh V.A. Immobilization of galactose
oxidase on parent and modified silica matrices // Abstr. 3th Int. Conf. Chemistry of highly
organized substances and scientific bases of nanotechnology. – St.-Petersburg, 2001. –
P.303-306.
L.V. Kondakova, V.V. Yanishpolskii, and V.A. Tertykh
L.V. Kondakova, V.V. Yanishpolskii, and V.A. Tertykh
L.V. Kondakova, V.V. Yanishpolskii, and V.A. Tertykh
L.V. Kondakova, V.V. Yanishpolskii, and V.A. Tertykh
Institute of Surface Chemistry, National Academy of Sciences
Gen. Naumov Str. 17, 03680 Kyiv-164, UKRAINE
Abstract
Introduction
Experimental
Results and discussion
Results and discussion
Results and discussion
Results and discussion
Results and discussion
Activity of adsorbed preparation,
Activity of adsorbed preparation,
AsilTDI-sample
Conclusions
Acknowledgement
References
|
| id | oai:ojs.pkp.sfu.ca:article-93 |
| institution | Surface |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2025-07-22T19:30:12Z |
| publishDate | 2002 |
| publisher | Chuiko Institute of Surface Chemistry National Academy of Sciences of Ukraine |
| record_format | ojs |
| resource_txt_mv | surfacezbircomua/56/7db4d23d466c33ab05403689b912b856.pdf |
| spelling | oai:ojs.pkp.sfu.ca:article-932018-11-27T09:42:19Z Adsorption and chemisorption of galactose oxidase on silica surface Adsorption and chemisorption of galactose oxidase on silica surface Adsorption and chemisorption of galactose oxidase on silica surface Kondakova, L. V. Yanishpolskii, V. V. Tertykh, V. A. Experience accumulated over a number of years in developing of methods of immobilization of galactose oxidase from Fusarium graminearum on parent and modified silica matrices is analyzed. Sturdy adsorption of galactose oxidase on silica surface was observed, such heterogeneous specimens possessed by enhanced biocatalyst stability and activity as compared with enzyme solutions. Covalent immobilization of galactose oxidase was carried out on the amine-containing silicas activated by 2,4-tolylene diisocyanate and cyanuric chloride. It was also shown that in the presence of the substrate (galactose) enzyme chemisorption takes place on the surface on amine-containing silica matrices. Immobilized preparations were successfully applied for analytical determination of galactose-containing carbohydrates (galactose, lactose, raffinose) in complex mixtures. Experience accumulated over a number of years in developing of methods of immobilization of galactose oxidase from Fusarium graminearum on parent and modified silica matrices is analyzed. Sturdy adsorption of galactose oxidase on silica surface was observed, such heterogeneous specimens possessed by enhanced biocatalyst stability and activity as compared with enzyme solutions. Covalent immobilization of galactose oxidase was carried out on the amine-containing silicas activated by 2,4-tolylene diisocyanate and cyanuric chloride. It was also shown that in the presence of the substrate (galactose) enzyme chemisorption takes place on the surface on amine-containing silica matrices. Immobilized preparations were successfully applied for analytical determination of galactose-containing carbohydrates (galactose, lactose, raffinose) in complex mixtures. Experience accumulated over a number of years in developing of methods of immobilization of galactose oxidase from Fusarium graminearum on parent and modified silica matrices is analyzed. Sturdy adsorption of galactose oxidase on silica surface was observed, such heterogeneous specimens possessed by enhanced biocatalyst stability and activity as compared with enzyme solutions. Covalent immobilization of galactose oxidase was carried out on the amine-containing silicas activated by 2,4-tolylene diisocyanate and cyanuric chloride. It was also shown that in the presence of the substrate (galactose) enzyme chemisorption takes place on the surface on amine-containing silica matrices. Immobilized preparations were successfully applied for analytical determination of galactose-containing carbohydrates (galactose, lactose, raffinose) in complex mixtures. Chuiko Institute of Surface Chemistry National Academy of Sciences of Ukraine 2002-06-12 Article Article application/pdf https://surfacezbir.com.ua/index.php/surface/article/view/93 Surface; No. 7-8 (2002): Chemistry, Physics and Technology of Surface; 150-157 Поверхность; № 7-8 (2002): Химия, физика и технология поверхности; 150-157 Поверхня; № 7-8 (2002): Хімія, фізика та технологія поверхні; 150-157 3154-8091 3154-8083 en https://surfacezbir.com.ua/index.php/surface/article/view/93/92 Авторське право (c) 2002 L.V. Kondakova, V.V. Yanishpolskii, V.A. Tertykh |
| spellingShingle | Kondakova, L. V. Yanishpolskii, V. V. Tertykh, V. A. Adsorption and chemisorption of galactose oxidase on silica surface |
| title | Adsorption and chemisorption of galactose oxidase on silica surface |
| title_alt | Adsorption and chemisorption of galactose oxidase on silica surface Adsorption and chemisorption of galactose oxidase on silica surface |
| title_full | Adsorption and chemisorption of galactose oxidase on silica surface |
| title_fullStr | Adsorption and chemisorption of galactose oxidase on silica surface |
| title_full_unstemmed | Adsorption and chemisorption of galactose oxidase on silica surface |
| title_short | Adsorption and chemisorption of galactose oxidase on silica surface |
| title_sort | adsorption and chemisorption of galactose oxidase on silica surface |
| url | https://surfacezbir.com.ua/index.php/surface/article/view/93 |
| work_keys_str_mv | AT kondakovalv adsorptionandchemisorptionofgalactoseoxidaseonsilicasurface AT yanishpolskiivv adsorptionandchemisorptionofgalactoseoxidaseonsilicasurface AT tertykhva adsorptionandchemisorptionofgalactoseoxidaseonsilicasurface |