RELEASE OF INTRACELLULAR β-GALACTOSIDASE FROM LACTOBACILLUS ACIDOPHILUS AND L-ASPARAGINASE FROM PECTOBACTERIUM CAROTOVORUM BY HIGH-PRESSURE HOMOGENIZATION

This article reports a comparison of the ease of disruption of gram-positive (Lactobacillus acidophilus) and gram-negative (Pectobacterium carotovorum) bacteria using high-pressure homogenization (HPH). The location factor for both enzymes calculated from enzyme release kinetics together with localization studies identified them as cytoplasmic enzymes. The results showed that release of β-galactosidase by HPH of L. acidophilus was more difficult than the release of L-asparaginase from P. carotovorum. It took nine passes at 55.14 MPa for maximum release of β-galactosidase (0.949 IU/mL) as compared to six passes at 41.35 MPa for L-asparaginase (4.653 IU/mL); 1.7 IU of β-galactosidase was released as against 11.5 IU of L-asparaginase per MJ of energy during high-pressure homogenization.

[1]  S. Harrison,et al.  Heat induced translocation of proteins and enzymes within the cell: an effective way to optimize the microbial cell disruption process , 2005 .

[2]  Yusuf Chisti,et al.  Disruption of microbial cells for intracellular products , 1986 .

[3]  S. Lele,et al.  Lactase Production from Lactobacillus acidophilus , 2005 .

[4]  A. Pandit,et al.  Significance of location of enzymes on their release during microbial cell disruption. , 2001, Biotechnology and bioengineering.

[5]  C. Michiels,et al.  High-Pressure Homogenization as a Non-Thermal Technique for the Inactivation of Microorganisms , 2006, Critical reviews in microbiology.

[6]  R. C. Dickson,et al.  Physiological studies of beta-galactosidase induction in Kluyveromyces lactis. , 1980, Journal of bacteriology.

[7]  T. Tsuchido,et al.  Heat‐Induced Translocation of Cytoplasmic β‐Galactosidase across Inner Membrane of Escherichiacoli , 1998, Biotechnology progress.

[8]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[9]  S. Harrison,et al.  The effect of chemical pretreatment combined with mechanical disruption on the extent of disruption and release of intracellular protein from E. coli , 2007 .

[10]  H. S. Choonia,et al.  RELEASE OF β-GALACTOSIDASE FROM INDIGENOUS LACTOBACILLUS ACIDOPHILUS BY ULTRASONICATION: PROCESS OPTIMIZATION , 2011 .

[11]  H. Verachtert,et al.  Localization and Characterization of α-Glucosidase Activity in Lactobacillus brevis , 1994 .

[12]  C. Michiels,et al.  Bacterial inactivation by high-pressure homogenisation and high hydrostatic pressure. , 2002, International journal of food microbiology.

[13]  Daniel G Bracewell,et al.  Advances in product release strategies and impact on bioprocess design. , 2009, Trends in biotechnology.

[14]  S. Harrison,et al.  Bacterial cell disruption: a key unit operation in the recovery of intracellular products. , 1991, Biotechnology advances.

[15]  S. Shifrin,et al.  L-Asparaginase from Erwinia carotovora. Physicochemical properties of the native and succinylated enzyme. , 1973, The Journal of biological chemistry.

[16]  M. Maillard,et al.  Efficient mechanical disruption of Lactobacillus helveticus, Lactococcus lactis and Propionibacterium freudenreichii by a new high-pressure homogenizer and recovery of intracellular aminotransferase activity , 2003, Journal of Industrial Microbiology and Biotechnology.

[17]  A. Middelberg,et al.  Process-scale disruption of microorganisms. , 1995, Biotechnology advances.

[18]  C. W. Robinson,et al.  Disruption of native and recombinant Escherichia coli in a high‐pressure homogenizer , 1989, Biotechnology and bioengineering.