Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Analysis of Kinetic Data of Pectinases with Substrate Inhibition

  • Gummadi, Sathyanarayana-N. (Biotechnology Programme, Department of Chemical Engineering, Indian Institute of Technology-Madras) ;
  • Panda, T. (Biotechnology Programme, Department of Chemical Engineering, Indian Institute of Technology-Madras)
  • Published : 2003.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

Enzyme kinetics data play a vital role in the design of reactors and control of processes. In the present study, kinetic studies on pectinases were carried out. Partially purified polymethylgalacturonase (PMG) and polygalacturonase (PG) were the two pectinases studied. The plot of initial rate vs. initial substrate concentration did not follow the conventional Michaelis-Menten kinetics, but substrate inhibition was observed. For PMG, maximum rate was attained at an initial pectin concentration of 3 g/l, whereas maximum rate was attained when the initial substrate concentration of 2.5 g/l of polygalacturonic acid for PG I and PG II. The kinetic data were fitted to five different kinetic models to explain the substrate inhibition effect. Among the five models tested, the combined mechanism of protective diffusion limitation of both high and inhibitory substrate concentrations (semi-empirical model) explained the inhibition data with 96-99% confidence interval.

Keywords

References

  1. Enzyme Microb. Technol. v.16 Concomitant synthesis of Aspergillus niger pectolytic enzymes and citric acid on sucrose Friedrich,J.;A.Cimerman;W.Steiner https://doi.org/10.1016/0141-0229(94)90093-0
  2. Bioproc. Eng. v.19 Production of pectolytic enzymes-a review Naidu,G.S.N.;T.Panda https://doi.org/10.1007/PL00009023
  3. Enzyme Microb. Technol. v.25 Performance of pectolytic enzymes during hydrolysis of pectic substances under assay conditions: A statistical approach Naidu,G.S.N.;T.Panda https://doi.org/10.1016/S0141-0229(99)00017-4
  4. Proc. Biochem v.38 Purification and biochemical properties of microbial pectinases: A review Gummadi,S.N.;T.Panda https://doi.org/10.1016/S0032-9592(02)00203-0
  5. Appl. Microbiol. Biotechnol. v.59 Microbial alkaline pectinases and their industrial applications: A review Hoordal,G.S.;R.P.Tiwari;R.Tewari;N.Dahiya;Q.K.Beg https://doi.org/10.1007/s00253-002-1061-1
  6. Bioresource Technol v.77 Application of pectinases in the commercial sector: A review Kashyap,D.R.;P.K.Vohra;C.Chopra;R.Tewari https://doi.org/10.1016/S0960-8524(00)00118-8
  7. Appl Microbiol. Biotechnol v.53 Perspectives in the biological function and the technological application of polygalacturonases Lang,C;H.Dornenburg https://doi.org/10.1007/s002530051628
  8. Proc. Biochem v.8 Bacteria, enzymes and wood permeability Fogarty,W.M.
  9. Understanding Enzymes Palmer,T.
  10. Biochim. Biophys. Acta v.1594 Substrate activation in acetylcholinesterase induced by low pH or mutation in the pi-cation subsite Masson,P.;L.M.Schopfer;C.F.Bartels;M.T.Froment;F.Ribes;F.Nachon;O.Lockridge https://doi.org/10.1016/S0167-4838(01)00323-5
  11. J. Enzyme Inhib. v.16 Slow detection reaction can mimic initial inhibition of an enzymic reaction Stojan,J. https://doi.org/10.1080/14756360109162358
  12. Biotechnol. Techniq. v.11 Biochemical determination of insecticides via cholinesterases;1. Acetylcholineesterase from rat brain: Functional expression using a baculovirus system and biochemical characterization Mionetto,N.;N.Morel;J.Massoulie;R.D.Schmid https://doi.org/10.1023/A:1018425224892
  13. Mol. Plant Microbe Interact v.8 Fungal polygalacturonases exhibit different substrate degradation patterns and differ in their susceptibilities to polygalacturonase-inhibiting proteins Cook,B.J.;R.P.Clay;C.W.Bergmann;P.Albersheim;A.G.Darvill
  14. Biotechnol. Bioeng. v.32 Batch kinetics of microbial polysaccharide biosynthesis Mulchandani,A.;J.H.T.Luong;A.Leduy https://doi.org/10.1002/bit.260320508
  15. Biotechnol. Bioeng. v.25 Growth kinetics of Dioscorea delltoidea and Catharanthus roseus in batch culture Drapeau,D.;H.W.Blanch;C.R.Wilke https://doi.org/10.1002/bit.260250220
  16. Proteinase Inhibitors Proteolytic enzymes Powers,J.C.;J.W.Harper;A.J.Barrett(ed.);G.Salvesin(ed.)
  17. Biological Chemistry Mahler,H.R.;E.H.Cordes
  18. The Bacteria Gunsalus,I.C.;R.Y.Stainer
  19. The Enzymes Boyer,P.D.;H.Lardy;K.Myrback
  20. The Enzymes Dixon,M.;Webb,E.C.
  21. Comprehensive Biochemistry Florkin,M.;E.H.Stotz
  22. Enzymes Haldane,J.B.S.
  23. Agric. Biol. Chem. v.30 Kinetic studies on microbial activities in concentrated solutions Ⅰ;Effect of excess sugars on oxygen uptake rate of a cell free respiratory system Yano,T.;T.Nakahara;S.Kamiyagam;K.Yamada https://doi.org/10.1271/bbb1961.30.42
  24. Enzyme and Metabolic Inhibitors Webb,J.L.
  25. Biotechnol. Bioeng. v.10 Kinetics of product inhibition in alcohol fermentation Aiba,S.;M.Shoda;M.Nagatani https://doi.org/10.1002/bit.260100610
  26. Biotechnol. Bioeng. v.27 Influence of high substrate concetration on microbial kinetics Edwards,V.H.
  27. Proc. Biochem. v.35 Multiresponse analysis of microbiological parameters affecting the production of pectolytic enzymes: A statistical view Naidu,G.S.N.;T.Panda https://doi.org/10.1016/S0032-9592(99)00051-5
  28. Bioproc. Eng. v.16 Statistical optimization of medium components for improved synthesis of pectinase by Aspergillus niger Nair,S.P.;T.Panda https://doi.org/10.1007/s004490050305
  29. J. Biol. Chem v.153 A photometric adaptation of the Somogyi method for the determination of glucose Nelson,N.
  30. J. Biol. Chem. v.195 Notes on sugar determination Somogyi,M.
  31. J. Biol. Chem. v.193 Protein measurement with Folin phenol reagent Lowry,O.H.;N.J.Rosenbrough;A.L.Farr;R.J.Randall
  32. Coll. Chechoslov. Chem. Comm. v.31 Isolation of extracellular pectolytic enzymes produced by Aspergillus niger Benkova,L.R.;A.Slezarik https://doi.org/10.1135/cccc19660122
  33. Response Surface Desingns and Analysis Khuri,A.I.;J.A.Cornell