Skip to main content
SpringerLink
Log in

Functional stabilization of invertase by covalent modification with pectin

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Pectin was attached to ethylenediamine-activated carbohydrate moieties of invertase using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide as coupling agent. The modified enzyme retained 57% of the original activity and contained 2.7 mol polymer per mol holoenzyme. Its optimum temperature was increased by 8 °C and its thermostability by 7.3 °C. The half-life at 65 °C was increased from 5 min to 2 days. The enzyme stability was enhanced by 33% at pH 2.0, and also by 27% at pH 12.0. The conjugate retained about 96% of its initial activity after 3 h incubation in 6 M urea.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anger H, Berth G (1986) Gel permeation chromatography and the Mark-Houwink relation for pectins with different degrees of esterification. Carbohydr. Polym. 6: 193–202.

    Google Scholar 

  • Bernfeld P (1955) Amylases α-and β-. Meth. Enzymol. 1: 149–158.

    Google Scholar 

  • Blomhoff HK, Christensen TB (1983) Effect of dextran and dextran modifications on the thermal and proteolytic stability of conjugated bovine testis β-galactosidase and human serum albumin. Biochim. Biophys. Acta 743: 401–407.

    PubMed  Google Scholar 

  • Bruneel D, Schacht E (1993) Chemical modification of pullulan: 2. Chloroformate activation. Polymer 34: 2633–2637.

    Google Scholar 

  • Dubois MK, Gilles A, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350–356.

    Google Scholar 

  • Gómez L, Ramírez HL, Villalonga R (2000) Stabilization of invertase by modification of sugar chains with chitosan. Biotechnol. Lett. 22: 347–350.

    Google Scholar 

  • Grasdalen H, Bakøy OE, Larsen B (1988) Determination of the degree of esterification and the distribution of methylated and free carboxyl groups in pectin by 1H-N.M.R. spectroscopy. Carbohydr. Res. 184: 183–191.

    Google Scholar 

  • Horovitz A, Serrano L, Avron B, Bycroft M, Fersht AR (1990) Strength and co-operativity of contributions of surface salt bridges to protein stability. J. Mol. Biol. 216: 1031–1044.

    PubMed  Google Scholar 

  • Husain S, Jafri F, Saleemuddin M (1996) Effects of chemical modification on the stability of invertase before and after immobilization. Enzyme Microb. Technol. 18: 275–280.

    Google Scholar 

  • Klibanov AM (1983) Stabilization of enzymes against thermal inactivation. Adv. Appl. Microbiol. 29: 1–28.

    PubMed  Google Scholar 

  • Pazur JH, Knull HR, Simpson DL (1970) Glycoenzymes: a note on the role for the carbohydrate moieties. Biochem. Biophys. Res. Commun. 40: 110–116.

    PubMed  Google Scholar 

  • Perutz ML (1978) Electrostatic effects in proteins. Science 201: 1187–1191.

    PubMed  Google Scholar 

  • Pollak A, Baughn RL, Adaisteinsson O, Whitesides GJ (1978) Immobilization of synthetically useful enzymes by condensation, polymerization. J. Am. Chem. Soc. 100: 302–304.

    Google Scholar 

  • Srivastava RAK (1991) Studies on stabilization of amylase by covalent coupling to soluble polysaccharides. Enzyme Microb. Technol. 13: 164–170.

    Google Scholar 

  • Trimble RB, Maley F (1977) Subunit structure of external invertase from Saccharomyces cerevisiae. J. Biol. Chem. 252: 4409–4412.

    PubMed  Google Scholar 

  • Villalonga R, Gómez L, Ramírez HL, Villalonga ML (1999) Stabilization of α-amylase by chemical modification with carboxymethylcellulose. J. Chem. Technol. Biotechnol. 74: 635–638.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Center for Biotechnological Studies, Faculty of Agronomy, University of Matanzas, Autopista a Varadero Km 3 1/2, Matanzas, C.P. 44740, Cuba

    Leissy Gómez & Reynaldo Villalonga

Authors
  1. Leissy Gómez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Reynaldo Villalonga
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gómez, L., Villalonga, R. Functional stabilization of invertase by covalent modification with pectin. Biotechnology Letters 22, 1191–1195 (2000). https://doi.org/10.1023/A:1005645531521

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005645531521

Search

Navigation