ZYMOGRAPHIC ANALYSIS OF LIPOLYTICAL ENZYMES OF SUBCELLULAR VIBRIO CHOLERAE FRACTIONS OF O1 AND O139 SEROGROUPS
Abstract and keywords
Abstract (English):
Zymographic analysis showed that all subcellular fractions of the serogroups possessed different degree of lipolytical activity. Maximal hydrolysis in the radial enzyme-diffusion reaction in agarose gel was observed in Tween-20, Tween-80, Triton Х-305 and Triton Х- 405. Total hydrolase activity of urea extracts in Tween-20, Tween-80, Triton Х- 405, Span-85 excelled those in the preparations of outer membranes. Maximal lipolytical activity in regard to the Tweens was shown by the preparations of urea extracts prepared from the nontoxigenic strains of O1 serogroups, while in relation to the Tritons – preparations of urea extracts from the toxigenic strains (p < 0,05).

Keywords:
zymographic analysis, subcellular fraction, lipolytical enzyme, Vibrio cholerae, substrate electrophoresis
References

1. Adamov AK (1986). Perspectives of cholera vaccine improvement [Perspektivy sovershenstvovaniya kholernykh vaktsin]. Epidemiologiya, epizootologiya i profilaktika osobo opasnykh infektsiy, 75-84.

2. Duvanova OV, Shimanyuk NY, Mishankin BN (2000). Ability to hydrolyze Tween-20 as a differential test for vibrios of O139 serovar of different origin [Sposobnost’ rasshcheplyat’ tvin 20 kak differentsial’nyy test dlya vibrionov O139 serovara razlichnogo proiskhozhdeniya]. Klinicheskaya laboratornaya diagnostika, (5), 48-49.

3. Kuzmichenko IA, Grachyova IV, Plotnikov OP (2001). Destructive activity and growth of Vibrio cholerae in presence of Tweens [Destruktivnaya aktivnost’ i rost kholernogo vibriona v prisutstvii tvinov]. Problemy osobo opasnykh infektsiy, 1 (81), 101-105.

4. Pakaneshchikova NV, Silishchev NN, Galimzyanova NF, Loginov ON (2006). Comparative study of dependence of lipolytical activity of Serratia genus bacteria on the content of nutritional media [Sravnitel’noe izuchenie zavisimosti lipoliticheskoy aktivnosti bakteriy roda Serratia ot sostava pitatel’noy sredy]. Bashkirskiy khimichesky zhurnal, 13 (4), 31-34.

5. Agafonova VV, Telesmanich NR, Akulova MV (2009). Method of differentiation of atoxigenic Vibrio cholerae strains of O1 and O139 serogroups from toxigenic strains by hydrolase activity: Patent N 2375457 of the Russian Federation [Sposob differentsiatsii atoksigennykh shtammov kholernykh vibrionov O1 i O139 serogrupp ot toksigennykh po gidrolaznoy aktivnosti: Pat. 2375457 Ros. Federatsiya: C1 C12Q 1/04, 1/30, C12R 1/63], (34), 8.

6. Kozlov SN, Markov EY, Nikolaev VB, Urbanovich LY, Zagoskina TY, Popova YO (2013). Method for determination of lipolytical activity in subcellular fractions of bacteria [Sposob opredeleniya lipoliticheskoy aktivnosti v subkletochnykh fraktsiyakh bakteriy: Pat. 2501014 Ros. Federatsiya; S1 G01N 33/50, 33/48], (34), 8.

7. Telesmanich NR (2004). Triacylglycerol lipase activity of haemolytic Vibrio cholerae [Triatsilglitserolipaznaya aktivnost’ gemoliticheskikh kholernykh vibrionov]. Zh. mikrobiol., epidemiol. i immunobiol., (2), 11-14.

8. Urbanovich LY, Markov EY, Golubinsky EP, Kolesnik RS, Sappo SG, Ganin VS, Karetnikova ES, Ivanova TA (1996). Characteristic of immunobiological properties of antigenic preparation of Vibrio cholerae outer membranes [Kharakteristika immunobiologicheskikh svoystv antigennogo preparata naruzhnykh membran kholernogo vibriona]. Biotekhnologiya, (6), 27-33.

9. Bornscheuer UT (2002). Microbial carboxyl esterases: classification, properties and application in biocatalysis. FEMS Microbiol. Rev., 26 (1), 73-81.

10. Hosseingholi EZ (2014). In silico analysis of Acinetobacter baumanri phospholipase B as a subunit vaccine candidate. Acta Biotheoretica, 64 (4), 455-475.

11. Ingole KV, Jalgaonkar SV, Fule RP (1998). Study of proteases and other enzymes of Vibrio cholerae O1 Eltor and O139 serotypes isolated in Yavatmal (Maharashtra). Indian J. Pathol. Microbiol., 41 (4), 419-422.

12. Jensen RG (1983). Detection and determination of lipase (acylglycerol hydrolase) activity from various sources. Lipids, 18 (9), 650-657.

13. Rosenstein R, Gotz F (2000). Staphylococcal lipases: biochemical and molecular characterization. Biochimie, 82, 1005-1014.

14. Schuepp C, Kermasha S, Michalski MC, Morin A (1997). Production, partial purification and characterisation of lipases from Pseudomonas fragi CRDA 037. Proc. Biohem., 32 (3), 225-232.

15. Stehr F, Kretschmar M, Kroger C, Hube B, Schafer W. (2003). Microbial lipases as virulence factors. J. Mol. Cat. B: Enzym., 22 (5-6), 347-355.

16. Whayeb SA, Yamamoto K, Castillo MY (1996). Lysophospholipase L2 of Vibrio cholerae O1 affects cholera toxin production. FEMS Immun. Med. Microbiol., 15 (1), 9-15.

Login or Create
* Forgot password?