Improving Properties of a Novel β-Galactosidase from Lactobacillus plantarum by Covalent Immobilization

Molecules

Rocio Benavente⁵  Benevides C. Pessela⁵  Jose Antonio Curiel¹  Blanca de las Rivas¹  Rosario Muñoz¹  Jose Manuel Guisán³  Jose M. Mancheño⁴  Alejandra Cardelle-Cobas²  Ana I. Ruiz-Matute⁶  Nieves Corzo⁶ 

[1] Grupo de Biotecnología Bacteriana, Instituto de Ciencia y Tecnología de Alimentos y Nutrición, (ICTAN-CSIC), Juan de la Cierva 3, 28006 Madrid, Spain; E-Mails:;CBQF—Centro de Biotecnologia e Química Fina, Escola Superior de Biotecnologia, Centro Regional do Porto da Universidade Católica Portuguesa, Rua Dr. António Bernardino Almeida, 4200-072 Porto, Portugal; E-Mail:;Departamento de Biocatálisis, Instituto de Catálisis y Petroleoquímica (ICP-CSIC), Marie Curie 2, Cantoblanco, 28049 Madrid, Spain; E-Mail:;Grupo de Cristalografía y Biología Estructural, Instituto de Química Física Rocasolano (IQFR-CSIC), Serrano 119, 28006 Madrid, Spain; E-Mail:;Departamento de Biotecnología y Microbiología de Alimentos, Instituto de Investigación en Ciencias de la Alimentación CIAL (CSIC-UAM), Campus de la Universidad Autónoma de Madrid, Nicolás Cabrera 9, 28049 Madrid, Spain; E-Mail:;Departamento de Bioactividad y Análisis de Alimentos, Instituto de Investigación en Ciencias de la Alimentación CIAL (CSIC-UAM), Campus de la Universidad Autónoma de Madrid, Nicolás Cabrera 9, 28049 Madrid, Spain; E-Mail:

关键词: β-galactosidase;    Lactobacillus plantarum;    immobilization;    glyoxyl-agarose;    oligosaccharides synthesis;    lactose;    lactulose;   

DOI  :  10.3390/molecules20057874

来源: mdpi

【 摘 要 】

A novel β-galactosidase from Lactobacillus plantarum (LPG) was over-expressed in E. coli and purified via a single chromatographic step by using lowly activated IMAC (immobilized metal for affinity chromatography) supports. The pure enzyme exhibited a high hydrolytic activity of 491 IU/mL towards o-nitrophenyl β-d-galactopyranoside. This value was conserved in the presence of different divalent cations and was quite resistant to the inhibition effects of different carbohydrates. The pure multimeric enzyme was stabilized by multipoint and multisubunit covalent attachment on glyoxyl-agarose. The glyoxyl-LPG immobilized preparation was over 20-fold more stable than the soluble enzyme or the one-point CNBr-LPG immobilized preparation at 50 °C. This β-galactosidase was successfully used in the hydrolysis of lactose and lactulose and formation of different oligosaccharides was detected. High production of galacto-oligosaccharides (35%) and oligosaccharides derived from lactulose (30%) was found and, for the first time, a new oligosaccharide derived from lactulose, tentatively identified as 3'-galactosyl lactulose, has been described.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.