Purification and Characterization of a Unique Alkaline Elastase from Micrococcus luteus

Clark, Deborah J.; Hawrylik, Steven J.; Kavanagh, Edward; Opheim, Dennis J.

doi:10.1006/prep.1999.1166

Cited by 16 publications

(12 citation statements)

References 45 publications

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“…There are also reports on novel keratinolytic actinobacteria assigned to genera Amycolatopsis or Actinomadura ( Al-Musallam et al , 2003 ; Puhl et al , 2009 ). The ubiquitous bacteria of the genus Micrococcus are known producers of extracellular serine or thiol proteases ( Mohedano et al , 1997 ; Odu and Akujobi, 2012 ), as well as elastases ( Clark et al , 2000 ). Nevertheless, the true keratinolytic potential of Micrococcus bacteria is sparsely mentioned and underestimated.…”

Section: Introductionmentioning

confidence: 99%

Keratinolytic abilities of <italic>Micrococcus luteus</italic> from poultry waste

Łaba¹,

Choińska²,

Rodziewicz³

et al. 2015

Braz. J. Microbiol.

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Keratinolytic microorganisms have become the subject of scientific interest due to their ability to biosynthesize specific keratinases and their prospective application in keratinic waste management. Among several bacterial classes, actinobacteria remain one of the most important sources of keratin-degrading strains, however members of the Micrococcaceae family are rarely scrutinized in regard to their applicatory keratinolytic potential. The tested Micrococcus sp. B1pz isolate from poultry feather waste was identified as M. luteus. The strain, grown in the medium with 1–2% chicken feathers and a yeast extract supplement, produced keratinases of 32 KU and lower level of proteases, 6 PU. It was capable to effectively decompose feathers or “soft” keratin of stratum corneum, in contrast to other “hard” hair-type keratins. The produced keratinolytic enzymes were mainly a combination of alkaline serine or thiol proteases, active at the optimum pH 9.4, 55 °C. Four main protease fractions of 62, 185, 139 and 229 kDa were identified in the crude culture fluid. The research on the auxiliary role of reducing factors revealed that reducing sulfur compounds could be applied in keratinolysis enhancement during enzymatic digestion of keratin, rather than in culture conditions. The presented M. luteus isolate exhibits a significant keratinolytic potential, which determines its feasible applicatory capacity towards biodegradation of poultry by-products or formulation of keratin-based feed components.

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Section: Introductionmentioning

confidence: 99%

Keratinolytic abilities of <italic>Micrococcus luteus</italic> from poultry waste

Łaba¹,

Choińska²,

Rodziewicz³

et al. 2015

Braz. J. Microbiol.

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“…It is crosslinked with nonpolar amino acid residues, such as Ala, Leu and Ile, and is combined with microfibrils to form elastic fibers in vivo [1,2]. Elastase is an endopeptidase that hydrolyzes insoluble elastin [3,4]. Because of its hydrolytic bias for peptide bonds at the carboxyl end of neutral aliphatic amino acids, elastase shows a high specificity for elastin.…”

Section: Introductionmentioning

confidence: 99%

“…However, the high cost of elastase sourced from animals has impeded its application. Microbial elastase serves as a preferred option not only because of the low cost and the extensive distribution of elastase in microorganisms, but also the ease to genetically manipulate enzyme properties for various requirements [3]. Since the first isolation and purification of elastase from Flavobacterium elastolyticum by Ines et al [13], elastase from Flavobacterium sp.…”

Section: Introductionmentioning

confidence: 99%

Isolation, identification and characterization of a novel elastase from Chryseobacterium indologenes

Lei

Zhao

et al. 2018

Appl Biol Chem

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Elastase is a type of protease that specifically degrades elastin. It has broad application prospects in medicine, food industry, and daily-use chemical industry. In this study, we isolated a bacterial strain WZE87 with high elastin-hydrolysis activity, which was identified as Chryseobacterium indologenes based on morphology, physiological and biochemical characteristics, and 16S rDNA sequence analysis. The elastase produced by this strain was purified by three steps: ammonium sulfate precipitation, Q-Sepharose fast-flow anion-exchange chromatography, and Sephadex G-75 gel-filtration chromatography. The purified elastase was 2376.5 U/mg in activity (a 8.3-fold increase in specific activity), and the recovery was 5.8%. Its molecular mass was estimated to be 26 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This enzyme was stable in the pH range of 5.0-10.5 at 37°C. The optimal temperature and pH were 37°C and 7.5, respectively. The activity of this elastase was found to decrease when the temperature was higher than 50°C. The activity was also inhibited by Zn 2? , Fe 2? , Fe 3? , and Mn 2? ions. The specific hydrolytic ability of this enzyme was similar to that of papain on substrates like gelatin, casein, soybean-isolated protein and bovine hemoglobin. However, this elastase preferentially hydrolyzed elastin in a protein mixture because of its specific adsorption. Considering its promising properties, this protease may be considered a potential candidate for applications in related industries.

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“…Survival potential in wide range of salt, pH and temperature and growth at high salt and alkaline pH makes Micrococcus an ideal candidate for study of industrial enzymes, especially alkaline protease. Although, a few studies related to alkaline protease production have already been done using some species of Micrococcus [8][9][10][11][12] but extensive data on screening, characterization and optimization of medium conditions and protease production processes using different strains of Micrococcus from diverse niches are still lacking. Our study focuses on isolation of halo-tolerant, efficient alkaline protease producing endophytic species of Micrococcus in comparison to reported species of the said genus.…”

Section: Introductionmentioning

confidence: 99%

Optimization of Nutrients and Culture Conditions for Alkaline Protease Production Using Two Endophytic Micrococci: Micrococcus aloeverae and Micrococcus yunnanensis

et al. 2017

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An endophytic species of was isolated from leaf (syn. ) and screened for protease production with five other species of. Data indicated that endophytic AE-6 MCC 2184 and DSM 21948 showed efficient protease production potential and secreted active protease at high salt (10%), temperature (40 °C) and in wide range of pH 8-10. Unlike . DSM 21948, protease production by . AE-6 MCC 2184 was stringently controlled by pH. Protease induction study using different group of peptides, peptide carbohydrates and peptide macronutrient combinations showed variable response with both the organisms. Result indicated that the amount of protease was not directly related to cell biomass but it depends on nature of inducible peptides. In this study we also developed a modified agar-well assay for semi-quantitative data from large number of replicates.

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Purification and Characterization of a Unique Alkaline Elastase from Micrococcus luteus

Cited by 16 publications

References 45 publications

Keratinolytic abilities of <italic>Micrococcus luteus</italic> from poultry waste

Keratinolytic abilities of <italic>Micrococcus luteus</italic> from poultry waste

Isolation, identification and characterization of a novel elastase from Chryseobacterium indologenes

Optimization of Nutrients and Culture Conditions for Alkaline Protease Production Using Two Endophytic Micrococci: Micrococcus aloeverae and Micrococcus yunnanensis

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