Dataset Information

Purification, Characterization and Thermodynamic Assessment of an Alkaline Protease by Geotrichum Candidum of Dairy Origin.

ABSTRACT: Background:Alkaline proteases is the important group of enzymes having numerous industrial applications including dairy food formulations. Objectives:The current study deals with the purification and characterization of an alkaline serine protease produced by Geotrichum candidum QAUGC01, isolated from indigenous fermented milk product, Dahi. Material and Methods:In total twelve G. candidum strains were screened for their proteolytic activity by using standard protease assay. The protease production from G. candidum QAUGC01 was optimized by varying physio-chemical conditions. The protease was purified by using two-step method: ammonium sulfate precipitation and gel filtration chromatography. Protease was further characterized by studying various parameter like temperature, pH, modulators, metal ions and organic solvent. A thermodynamic study was also carried out to explore the half-life of protease. Results:The G. candidum grew profusely at 25 °C and at an initial pH of 4.0 for 72 h of incubation producing 26.21 U/ml maximum extracellular protease. Protease revealed that Vmax and Km was 26.25 U.ml-1.min-1 and 0.05 mg.mL-1, respectively using casein as substrate. The enzyme was stable at a temperature range (25-45 °C) and pH (8-9). Residual enzyme activity was strongly inhibited in the presence of PMSF (7.5%). The protease could hydrolyze proteinaceous substrates, casein (98%) and BSA (95%). The thermodynamic studies explored that the half-life of the enzyme that was 106.62 min, 38.72 min and 15.71 min at 50, 60 and 70 °C, respectively. Conclusions:Purified protease from G. candidum GCQAU01 is an ideal candidate for industrial application.

SUBMITTER: Muhammad A

PROVIDER: S-EPMC6697849 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Purification, Characterization and Thermodynamic Assessment of an Alkaline Protease by <i>Geotrichum Candidum</i> of Dairy Origin.

Muhammad Abubakar A Bokhari Syed Ali Imran SAI Vernoux Jean-Paul JP Ali Muhammad Ishtiaq MI Faryal Rani R Desmasures Nathalie N Imran Muhammad M

Iranian journal of biotechnology 20190420 2

<h4>Background</h4>Alkaline proteases is the important group of enzymes having numerous industrial applications including dairy food formulations.<h4>Objectives</h4>The current study deals with the purification and characterization of an alkaline serine protease produced by <i>Geotrichum candidum</i> QAUGC01, isolated from indigenous fermented milk product, Dahi.<h4>Material and methods</h4>In total twelve <i>G. candidum</i> strains were screened for their proteolytic activity by using standard ...[more]

PMID: 31457056

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Project description:The yeast Geotrichum candidum (teleomorph Galactomyces candidus) is inoculated onto mold- and smear-ripened cheeses and plays several roles during cheese ripening. Its ability to metabolize proteins, lipids, and organic acids enables its growth on the cheese surface and promotes the development of organoleptic properties. Recent multilocus sequence typing (MLST) and phylogenetic analyses of G. candidum isolates revealed substantial genetic diversity, which may explain its strain-dependant technological capabilities. Here, we aimed to shed light on the phenotypic and genetic diversity among eight G. candidum and three Galactomyces spp. strains of environmental and dairy origin. Phenotypic tests such as carbon assimilation profiles, the ability to grow at 35°C and morphological traits on agar plates allowed us to discriminate G. candidum from Galactomyces spp. The genomes of these isolates were sequenced and assembled; whole genome comparison clustered the G. candidum strains into three subgroups and provided a reliable reference for MLST scheme optimization. Using the whole genome sequence as a reference, we optimized an MLST scheme using six loci that were proposed in two previous MLST schemes. This new MLST scheme allowed us to identify 15 sequence types (STs) out of 41 strains and revealed three major complexes named GeoA, GeoB, and GeoC. The population structure of these 41 strains was evaluated with STRUCTURE and a NeighborNet analysis of the combined six loci, which revealed recombination events between and within the complexes. These results hint that the allele variation conferring the different STs arose from recombination events. Recombination occurred for the six housekeeping genes studied, but most likely occurred throughout the genome. These recombination events may have induced an adaptive divergence between the wild strains and the cheesemaking strains, as observed for other cheese ripening fungi. Further comparative genomic studies are needed to confirm this phenomenon in G. candidum. In conclusion, the draft assembly of 11 G. candidum/Galactomyces spp. genomes allowed us to optimize a genotyping MLST scheme and, combined with the assessment of their ability to grow under different conditions, provides a reliable tool to cluster and eventually improves the selection of G. candidum strains.

Dataset Information

Purification, Characterization and Thermodynamic Assessment of an Alkaline Protease by Geotrichum Candidum of Dairy Origin.

Publications

Purification, Characterization and Thermodynamic Assessment of an Alkaline Protease by <i>Geotrichum Candidum</i> of Dairy Origin.

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