Production, purification, and characterization of a polygalacturonase from a new strain of kluyveromyces marxianus isolated from coffee wet-processing wastewater |
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Authors: | Manuel Serrat Rose Catalina Bermúdez Tomás Gonzáles Villa |
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Institution: | (1) Centro de Estudios de Biotecnología Industrial, Universidad de Oriente, Patricio Lumumba s/n, 90500 Santiago de Cuba, Cuba;(2) Facultad de Farmacia, Cátedra de Microbiología, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain |
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Abstract: | A new high polygalacturonase (PG)-producing Kluyveromyces marxianus strain was isolated from coffee wet-processing wastewater. PG production in this strain is not repressed in the presence
of 100g/L of glucose and, being growth-associated, reached its maximum accumulation in the culture medium at the beginning
of the stationary phase. Oxygen and galacturonic acid negatively regulated enzyme synthesis, and glucose as the carbon source
afforded better enzyme yields than lactose. The data reported here show that this strain exhibits the highest index of PG
production among the wild-type strains reported so far (18.8U/mL). PG was readily purified by ion-exchange chromatography
on SP-Sepharose FF. The activity corresponded to a single protein with an M
r of 41.7 kDa according to sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was stable in the pH range
of 3.0–5.0 and displayed an optimal temperature of 55°C; it showed a typical endo-splitting way of substrate hydrolysis and exhibited a fair degree of activity on pectin with a high degree of esterification. |
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Keywords: | Polygalacturonase pectin production characterization Kluyveromyces marxianus |
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