Production of Geotrichum candidum polygalacturonases via solid state fermentation on grape pomace
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Kateřina Illková
,
Zuzana Zemková
Abstract
Geotrichum candidum CCY 16-1-29 (teleomorph Galactomyces geotrichum) is able to grow and produce polygalacturonase of remarkable activities on pectin or grape pomace as a sole carbon source. The highest activities of extracellular enzymes were found on the third and the seventh day of cultivation. After extraction and precipitation, polygalacturonases produced in these cultivation periods were characterized. Production of multiple forms of polygalacturonase was observed in both cultivation periods. Two major forms, polygalacturonase with random action pattern (endo-PGase, EC 3.2.1.15) and oligogalacturonate hydrolase (exoPGase, exopolygalacturonase preferring oligogalacturonides as substrates), as well as numerous minor forms were detected by IEF-PAGE using the print technique detection. EndoPGase was identified by mass spectrometry. The major forms have similar isoelectric points (below pH 6.0) and pH optima (4.6 and 4.8, respectively). pH optimum of 4.6 was associated with exoPGase and that of 4.8 with endoPGase. Both enzymes were stable after freeze-drying and storage at 4°C. EndoPGase had molecular mass of about 29 kDa (36 kDa by SDS-PAGE) as determined by gel filtration, temperature optimum of about 45°C and it was stable only below 35°C. Molecular mass of exoPGase was about 50 kDa, its temperature optimum was about 60°C, and it was stable to 60°C. Optimal substrate for exoPGase was a pentamer, for endoPGase it was a pectate. Values of K m for optimal substrate reached the values of 11.4 × 10−5 M for for exoPGase and 6.6 × 10−5 M for endoPGase. Pectin methylesterase as another pectolytic enzyme was also identified by mass spectrometry.
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© 2012 Institute of Chemistry, Slovak Academy of Sciences
Artikel in diesem Heft
- 5th Meeting on Chemistry & Life 2011
- Induction of Cryptococcus laurentii α-galactosidase
- Incorporation of β-(1,6)-linked glucooligosaccharides (pustulooligosaccharides) into plant cell wall structures
- Metal-metabolomics of microalga Chlorella sorokiniana growing in selenium- and iodine-enriched media
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- Seasonal changes of Rubisco content and activity in Fagus sylvatica and Picea abies affected by elevated CO2 concentration
- Identification and determination of relatedness of lactobacilli using different DNA amplification methods
- Production of Geotrichum candidum polygalacturonases via solid state fermentation on grape pomace
- Monitoring of yeast population isolated during spontaneous fermentation of Moravian wine
- Biodegradable polyhydroxybutyrate as a polyol for elastomeric polyurethanes
- Conformational changes in humic acids in aqueous solutions
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Artikel in diesem Heft
- 5th Meeting on Chemistry & Life 2011
- Induction of Cryptococcus laurentii α-galactosidase
- Incorporation of β-(1,6)-linked glucooligosaccharides (pustulooligosaccharides) into plant cell wall structures
- Metal-metabolomics of microalga Chlorella sorokiniana growing in selenium- and iodine-enriched media
- Metabolomic approach to Alzheimer’s disease diagnosis based on mass spectrometry
- Seasonal changes of Rubisco content and activity in Fagus sylvatica and Picea abies affected by elevated CO2 concentration
- Identification and determination of relatedness of lactobacilli using different DNA amplification methods
- Production of Geotrichum candidum polygalacturonases via solid state fermentation on grape pomace
- Monitoring of yeast population isolated during spontaneous fermentation of Moravian wine
- Biodegradable polyhydroxybutyrate as a polyol for elastomeric polyurethanes
- Conformational changes in humic acids in aqueous solutions
- Preparation and properties of cementitious composites for geothermal applications
