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ISSN 1998-4812
Вестник Башкирского университета. 2015. Т. 20. №4
1177
y^K 541.64:536
Краткое сообщение
INFLUENCE OF SOME ANTIBIOTICS OF A CEPHALOSPORIN SERIES ON THE PROCESS OF ENZYME HYDROLYSIS OF CHITOZAN IN THE SOLUTION OF ACETIC ACID
© A. S. Shurshina, M. R. Garifullina, M. A. Elinson, I. F. Tuktarova, V. V. Chernova*
Bashkir State University 32 Zaki Validi St., 450076 Ufa, Republic of Bashkortostan, Russia.
*Email: [email protected] The process of enzyme hydrolysis of chitosan in the presence of some antibiotics ofa cephalosporin series - sodium salt of cefazolin and cefatoksim is studied by the method of viscometry. It is established that the introduction of medicines in the solution of chitosan leads to the reduction of a maximum rate of enzyme hydrolysis Vmax and to increase a constant of Mikhaelis Km that points the possibility of a slow transformation of a substratum in a threefold complex of chitosan-enzyme-antibiotic and allows to describe an effect of cefazolin and a cefatoksim within the model of a mixed inhibition. Keywords: chitosan, viscosity, enzyme hydrolysis.
Introduction
Nowadays one of the perspective directions of the use of chitosan (ChT) is the creation of the film materials on its basis including drugs for the treatment of surgical, burnt and sore wounds. In the conditions of medical application of chi-tosan materials their biodestruction is carried out under the influence of nonspecific enzymes of a human body (for example, hyaluronidase) and defines covering service life on a wound surface. In the works of previous years we defined some kinetic characteristics of the activity of enzyme hya-luronidase for the reaction of enzyme hydrolysis of ChT in the solution of 1% acetic acid [1], and also in the presence of antibiotics of an aminoglycoside series - amikacin [2, 3] and gentamycin [4]. This work is undertaken with the purpose of the expansion of some medical substances (MS) immobilized in a polymeric matrix.
Experimental part
The following objects were used in our research: they were ChT of "Bioprogress" enterprise (Russia) with a molecular mass of Msn=113000, antibiotics of cephalosporin series - sodium salt of cefazolin (CFZ) and cefatoksim (CFT) produced by "Sintez" enterprise (Russia), an enzyme preparation hyaluronidase (a trade name of the preparation is "Liraz") produced by "Mikrogen" (Russia). Concentration of enzyme preparation was 0.1 g/l. 1% acetic acid was used as some solvent for ChT. Concentration of ChT in the solution for carrying out the process of enzyme hydrolysis varied from 0.1 to 5 g/dl. Kinetic constants of the process of enzyme hydrolysis were defined according to the method described in the works [1-4] at molar ratios of ChT:MS. It was equal to 1:0.01 and 1:0.1.
Discussion of the results
The conducted researches showed that the observed dependences of an initial rate of enzyme hydrolysis of ChT in the presence of CFZ and CFT from concentration of a substratum (ChT) can be described within the scheme of Mi-khaelisa-Menten, as well as in case of enzyme hydrolysis of ChT with the lack of drugs [1]. The results of Mikhaelis constant, the maximum rate determined by a graphic method of Laynuivera-Burke [5] are presented in table 1.
Table 1
The value of kinetic parameters of the process of enzyme hydrolysis in Mikhaelisa-Menten equation for the system
Molar
ratio of
Used MS components ChT : MS Km, g/dl Vmax-106, g/(dlmin) Vmax/Km'106, min-1
CFZ 1:0.01 3.82 0.47 0.12
1:0.10 3.91 0.40 0.10
CFT 1:0.01 3.79 0.48 0.13
1:0.10 3.95 0.41 0.10
The received values of Km for the systems of ChT:CFZ and ChT:CFT testify that the affinity of ChT to enzyme in the presence of antibiotics worsens. Besides the reduction values of Vmax/Km in the systems of ChT-MS in comparison with an individual ChT says that drugs systems are more steady to biodegradation processes.
Thus, according to the influence of CFZ and CFT on the process of enzyme hydrolysis of ChT, antibiotics of a cephalosporin series, practically don't differ from the antibiotics of an aminoglycoside series which had been researched earlier. It is obviously connected with the fact that all of them represent low-molecular electrolytes.
REFERENCES
1. Kulish E. I., Tuktarova I. F., Chernova V. V., Kolesov S. V. Studying of the process of enzyme destruction of chitosan in the solution of acetic acid // Bulletin of the Bashkir University. 2013. T. 18. №3. P. 688-690.
2. Chernova V. V., Shurshina A. S., Bazunova M. V., Kulish E. I. Studying of enzyme hydrolysis of chitosan in the solution of acetate buffer // Journal of applied chemistry. 2015. T. 88. N°. 4. P. 616-621.
3. Kulish E. I., Tuktarova I. F., Chernova V. V. Enzyme stability of drugs film materials on the basis of chitosan // Perspective Materials. 2014. №6. P. 25-31.
4. Tuktarova I. F., Lazdin R. Yu., Chernova V. V. Studying of the influence of an antibiotic gentamicin sulfate on the process of enzyme hydrolysis of chitosan in the solution of acetic acid // Scientific and technical bulletin of the Volga region. 2015. №2. P. 41-43.
5. Biochemistry/ edit E. S. Severin. M.: Geotard-Med, 2004. 779 p.
Received 14.11.2015.
