β-Galactosidase from Kluyveromyces lactis cell disruption and enzyme immobilization using a cellulose–gelatin carrier system

Abstract Whole cell immobilization is one of the major immobilization methods due to cost advantages. Extracellular enzyme producing cells can be immobilized directly, but intracellular enzyme producing cells should be treated first to increase cell permeability. In this work β-galactosidase-producing Kluyveromyces lactis (ATCC 8583) cells were used. Since β-galactosidase is an intracellular enzyme permeabilized dead cells were immobilized into gelatin using glutaraldehyde as cross-linker. Two chemical and one physical disruption processes were tested and the physical method was determined to be better because of the probable risk of chemical toxicity accompanied with the chemical methods. Thirty percent activity was obtained by immobilized cells relative to free disrupted cells.

[1]  S. Shimizu,et al.  Hydrolysis of lactose by immobilized microorganisms , 1977, Applied and environmental microbiology.

[2]  Channing R. Robertson,et al.  The immobilization of whole cells: Engineering principles , 1985 .

[3]  Ural Akbulut,et al.  Immobilisation of β‐galactosidase onto gelatin by glutaraldehyde and chromium(III) acetate , 1994 .

[4]  J. Teissié,et al.  Electroinduced extraction of β-galactosidase from Kluyveromyces lactis , 2001, Applied Microbiology and Biotechnology.

[5]  U. Akbulut,et al.  Immobilization of Urease into Carboxymethylcellulose - Gelatine System , 1992 .

[6]  K. Yam,et al.  Effects of Individual Components in a Vitamin E Formulation on Off-Odor Release, Melt Flow Index, and Yellowness Index of an Hdpe Polymer , 1999 .

[7]  R. R. Mahoney,et al.  PURIFICATION AND CHARACTERIZATION OF β‐GALACTOSIDASE FROM MUCOR PUSILLUS , 1997 .

[8]  M. Kula,et al.  Purification of Proteins and the Disruption of Microbial Cells , 1987 .

[9]  J. Cabral,et al.  Penicillin acylase release from Escherichia coli cells by mechanical cell disruption and permeabilization , 2002 .

[10]  Nikos G. Papayannakos,et al.  Studies on modelling and simulation of lactose hydrolysis by free and immobilized β-galactosidase from Aspergillus niger , 1993 .

[11]  M. Becerra,et al.  Yeast β-galactosidase in solid-state fermentations , 1996 .

[12]  P. Rouxhet,et al.  Immobilization of lactase in yeast cells retained in a glass wool matrix , 1989 .

[13]  S. Furlan,et al.  Optimization of pH, Temperature and Inoculum Ratio for the Production of β‐D‐Galactosidase by Kluyveromyces marxianus Using a Lactose‐free Medium , 2001 .

[14]  B. Brena,et al.  Activity and stability of Escherichia coli β-galactosidase in cosolvent systems , 1998 .

[15]  E. L. Harris,et al.  Protein purification applications : a practical approach , 1990 .

[16]  V. Gekas,et al.  Hydrolysis of lactose: a literature review , 1985 .

[17]  E. Šturdı́k,et al.  Optimization of β-galactosidase extraction from Kluyveromyces marxianus , 1993 .

[18]  Jiaye Yu,et al.  L‐Asparaginase Release from Escherichia coli Cells with K2HPO4 and Triton X100 , 2001, Biotechnology progress (Print).

[19]  Robert Eisenthal,et al.  Enzyme assays : a practical approach , 1992 .

[20]  J. Whitaker,et al.  PURIFICATION AND PHYSICOCHEMICAL PROPERTIES OF ?-GALACTOSIDASE FROM Kluyveromyces fragilis , 1978 .