Purification and characterization of extracellular β-galactosidase from the psychrotolerant yeast Guehomyces pullulans 17-1 isolated from sea sediment in Antarctica |
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| Institution: | 1. Laboratory of Food Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University & Research, Bornse Weilanden 9, 6708WG Wageningen, the Netherlands;2. Synthetic and Systems Biology Unit, Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Temesvári krt. 62, H-6726 Szeged, Hungary;3. Department of Bioengineering, University of California, 9500 Gilman Drive La Jolla, San Diego, CA 92093, USA;4. Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800 Kgs. Lyngby, Denmark;1. State Key Laboratory of Bioreactor Engineering, Department of Food Science and Technology, School of Biotechnology, East China University of Science and Technology, Shanghai 200237, People’s Republic of China;2. State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, People’s Republic of China;3. Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), Shanghai 200237, People’s Republic of China;1. Instituto de Ciencias Biológicas, Universidad de Talca, Avda. Lircay s/n, Talca, Chile;2. Núcleo Científico Multidisciplinario-DI, Universidad de Talca, Avda. Lircay s/n, Talca, Chile;3. Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Pozuelo de Alarcón, Spain;4. Centro de Estudios Fotosintéticos y Bioquímicos, Universidad Nacional de Rosario, Rosario, S2002LRK, Argentina;5. Centro de Estudios en Ecología Molecular y Funcional, Universidad de Talca, Talca, Chile;6. Centro de Estudios Avanzados en Zonas Áridas (CEAZA), Universidad Católica del Norte, Coquimbo, Chile;1. Departamento de Microbiologia, Universidade Federal de Viçosa, Viçosa, MG, Brazil;2. Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, MG, Brazil;3. Núcleo de Análise de Biomoléculas (NuBioMol), Universidade Federal de Viçosa, Viçosa, MG, Brazil;4. Departamento de Biologia Vegetal, Universidade Federal de Viçosa, Viçosa, MG, Brazil;1. Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China;2. State Key Laboratory of Chemical Engineering, Tianjin University, Tianjin 300072, PR China;3. Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, PR China;4. Tianjin Key Laboratory of Membrane Science and Desalination Technology, Tianjin University, Tianjin 300072, PR China |
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| Abstract: | A psychrotolerant yeast Guehomyces pullulans 17-1 isolated from sea sediment in Antarctica could produce high level (17.2 U/ml) of both extracellular and cell-bound β-galactosidase. The extracellular β-galactosidase in the supernatant of the cell culture of the psychrotolerant yeast G. pullulans 17-1 was purified to homogeneity with a 2.4-fold increase in specific activity as compared to the supernatant by concentration, gel filtration chromatography (Sephadex G-200) and cation-exchange chromatography (CM-Sepharose Fast Flow cation-exchange). The molecular mass of the purified extracellular β-galactosidase was estimated to be 335 kDa. The optimal temperature and pH of the purified β-galactosidase were 50 °C and 4.0, respectively. Km and Vmax values of the purified β-galactosidase for o-nitrophenyl-β-d-galactopyranoside were 3.3 mM and 9.2 μmol/min. Lactose can be converted into glucose and galactose and a large amount of reducing sugar can be released from milk under catalysis of the purified β-galactosidase. The matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectroscopy identified a peptide ALEEYKK which is the conserved motif of the β-galactosidases from other yeasts. The results show that the enzyme may have potential applications in food industry. |
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