• Journal of Microbiology and Biotechnology
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• v.23 no.7
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• pp.1004-1014
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• 2013

Six nonpathogenic fungal strains isolated from alkaline soils of Buenos Aires Province, Argentina (Acremonium murorum, Aspergillus sidowii, Cladosporium cladosporoides, Neurospora tetrasperma, Purpureocillium lilacinum (formerly Paecilomyces lilacinus), and Westerdikella dispersa) were tested for their ability to produce keratinolytic enzymes. Strains were grown on feather meal agar as well as in solid-state and submerged cultures, using a basal mineral medium and "hair waste" as sole sources of carbon and nitrogen. All the tested fungi grew on feather meal agar, but only three of them were capable of hydrolyzing keratin, producing clear zones. Among these strains, P. lilacinum produced the highest proteolytic and keratinolytic activities, both in solid-state and submerged fermentations. The medium composition and culture conditions for the keratinases production by P. lilacinum were optimized. Addition of glucose (5 g/l) and yeast extract (2.23 g/l) to the basal hair medium increased keratinases production. The optimum temperature and initial pH for the enzyme production were $28^{\circ}C$ and 6.0, respectively. A beneficial effect was observed when the original concentration of four metal ions, present in the basal mineral medium, was reduced up to 1:10. The maximum yield of the enzyme was 15.96 $U_c/ml$ in the optimal hair medium; this value was about 6.5-fold higher than the yield in the basal hair medium. These results suggest that keratinases from P. lilacinum can be useful for biotechnological purposes such as biodegradation (or bioconversion) of hair waste, leading to a reduction of the environmental pollution caused by leather technology with the concomitant production of proteolytic enzymes and protein hydrolyzates.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.4
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• pp.594-599
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• 2001

This study was attempted to isolate and identify the strain revealing high angiotensin converting enzyme (ACE) inhibitory activity from various soy fermented foods, i.e. meju, soybean paste and soy sauce. Forty-two strains with morphologically different characteristics were selected and the ACE inhibitory and proteolytic activities were examined. Of the strains tested, SS103 which was isolated from soy sauce showed the highest ACE inhibitory and proteolytic activities and was finally selected for further studies. The SS103 strain showed motility, rod form and ellipsoidal spores. The shape of colonies on the agar media was irregular, mucoidal and surface dull. The strain could grow under aerobic conditions of pH 5~9 and 10~$50^{\circ}C$. Main cellular fatty acid was $C_{15:0}$ anteiso, $C_{17:0}$ cis and $C_{17:0}$ iso, which was 33.9%, 18.8% and 16.5%, respectively. Based upon these morphological, biochemical and cultural properties, SS103 was identified as a Bacillus subtilis. Optimum cultural condition of Bacillus subtilis SS103 was pH 8.0, $37^{\circ}C$ and 48 hr.48 hr.

• Korean Journal of Microbiology
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• v.30 no.2
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• pp.71-77
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• 1992

A 50 KD metalloprotease of Serratia marcesrens ATCC 21074 was purified by ammoniumsulfate precipitation. DEAE-cellulose ion exchange chromatography, and sephadex ti-100gel filtration. Optimal pH and temperature of enzyme were pH 8.0 and 37"C, respectively.This enzyme was stable in the ranges of 10-37$^{\circ}$C and pH 5.0--11.0. Thermal denaturationwas investigated by differential scanning calorimetry. Onset temperature of denaturationand endothermic peak temperature were 376$^{\circ}$C and 43.2"C. re:,pectively. The denaturationenthalpy was -8.4mJimg. The purified metalloprotease was ri.sistant to autodigestion for24 hr at 30$^{\circ}$C. Metalloprotease in culture supernatant was also resistant to autodigestionin this conditions. Heat-denatured enzyme. however. was rapidly digested by the nativeenzyme. The metalloprotease was stable to proteolytic digestion by mammalian proteasessuch as trypsin. a-chymotrypsin, and elastase. But the enzyme was easily digested bybacterial protease. thermolysin.bacterial protease. thermolysin.

• The Journal of the Korean dental association
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• v.11 no.8
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• pp.525-530
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• 1973

True collagenolytic enzymes in animal tissues were first demonstrated by Gross and Lapiere (1962), who showed the ability of such an enzyme in the culture medium of living explants of tadpole tissue to degrade a specific substrate of undenatured collagen under physiological conditions. Recently, tumor-associated collagenolytic activity has been demonstrated in human neoplasm and in ascites V Carcinoma. This investigation have been peforme to determine whether or not a collagen lytic enzyme could e found in isolated solid Tawa sarcoma of Donryu female rat obtained the culture medium. The results were as follows. 1. 11.5mg% of hydroxyproline contained in Donryu rat skin collagen, which was extracted by 0.5M acetic acid. 2. Cultivation of solid Tawa sarcoma tissues on reconstituted rat skin collagen gels showed lysis of adjacent gel after 18 hours, and much more extensive lysis after 5 days. 3. Collagen substrate was not attacked by the common proteolytic enzymes, trypsin, pepsin, and pronase.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.189-196
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• 1997

A producer of inhibitor against ${\beta}-glucosidase$ of Penicillium sp.-L4 was screened from Actinomycetes, and the isolated strain was identified as Streptomyces sp. The inhibitor produced was very stable against heat, acidic and alkaline conditions, proteolytic and amylolytic enzymes. The inhibotor was purified from culture broth through activated carbon treatment, ultrafiltration, anion and cation exchange, activated carbon columm, acetone precipitation and preparative HPLC. It showed inhibitory activities against a variety of dissacharide hydrolyzing enzymes produced by P.sp.-L4, and the mode of inhibition was competitive. Its structure and molecular formular was elucidated by IR, $^1H\;and\;^{13}C$ NMR and FAB/Mass spectrometry, which was identified as 1-deoxynojirimycin (dNM). dNM showed inhibitory effects on the cell growth and hydrolytic enzyme action of P.sp.-L4 on agar plate and infected lemon peel.

• Biomedical Science Letters
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• v.10 no.4
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• pp.347-351
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• 2004

Maggot fibrolase (MsMg-1) was purified from the maggots of Mimela splendems using ammonium sulfate fractionation, DEAE Affi-gel affinity chromatography. This protease had a molecular weight of 85 kDa as determined by SDS-polyacrylarnide gel electrophoresis under reducing conditions. It showed strong proteolytic and fibrinolytic activities. The purified enzyme was strongly inhibited by phenylmethanesulfonyl fluoride, Mn/sup 2+/, and Zn/sup 2+/ but it was not by EDTA, EGT, Mg/sup 2+/, Ca/sup 2+/, and Li/sup 2+/ ions. In these experimental results, we have speculated that MsMg-1 is a serine protease with a strong fibrinolytic activity.

• Journal of Life Science
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• v.11 no.2
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• pp.138-143
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• 2001

Marsh calm(Corbicular elatior)with a short-term storage in raw and a law-rate of utilization has been increasing the needs to develop new marsh calm processing products for a temporary mass treatment and long-term distribution, Therefore the processing conditions of marsh calm beverage using proteolytic enzyme hydrolysis were investigated. A partial hydrolysis at 6$0^{\circ}C$ for 1 hour after adding 3% Alcalase as more effective than a hot water extraction to develop taste compounds from the marsh calm. The result of ommission test showed that nucleotides and their related compounds were contributed in the taste of the marsh calm hydrolysates rather than free amino acids. The taste of the hydrolysates was produced by association with these compounds rather than only one compound s the hydrolystes taste differently for the control when one of these compound was omitted. The hydrolysates were fractionated to molecular weight below 500 dalton to eliminate bitter taste and to improve it flavor from the hydrolysates, 0.05% bay leaf was more effective to improve the odor than other herbs.

• BMB Reports
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• v.41 no.12
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• pp.827-832
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• 2008

Many age-dependent neurodegenerative diseases are associated with the accumulation of abnormally folded proteins within neurons. One of the major proteolytic pathways in the cell is the autophagy pathway, which targets cytoplasmic contents and organelles to the lysosomes for bulk degradation under various physiological and stressful conditions. Although the importance of autophagy in cellular physiology is well appreciated, its precise roles in neurodegeneration remain largely unclear. Recent studies indicate that components of the endosomal sorting complex required for transport (ESCRT) are important in the autophagy pathway. Reduced activity of some ESCRT subunits leads to the accumulation of autophagosomes and failure to clear intracellular protein aggregates. Interestingly, rare mutations in CHMP2B, an ESCRT-III subunit, are associated with frontotemporal dementia linked to chromosome 3 (FTD3). Mutant CHMP2B proteins seem to disrupt the fusion of autophagosomes and lysosomes in cell culture models. These findings suggest a potential mechanism for the pathogenesis of FTD3 and possibly other neurodegenerative diseases as well.

• Korean Journal of Microbiology
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• v.10 no.2
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• pp.79-86
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• 1972

The possibility of substituting potato or sweet potato for the wheat, one of the raw materials for soy sauce, was studied by measuring the amylase and proteolytic activities of Koji. Also optimum conditions of Koji making were determined. It was found that substitution of up to 30% (starch content) of wheat content (15% of the total bean and wheat content) with potato yielded good qulity of soy sauce. Use of more than 30% potato yielded a Koji of low enzymatic activity. This was attributed to the high moisture content of potato. It ws also found that substitution of up to 50% (starch content) of wheat content(25% of the total bean and wheat content) with sweet potato yieldede a good quality of soy sauce. But the taste was inferior to the control (the soy sauce which was made with 50% of bean and 50% of wheat).

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.1
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• pp.7-13
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• 2001

Enzymatic hydrolysis of different forms of silk fibroin (soluble, gel and insoluble forms) by industrial and commercial enzyme preparations to obtain aqueous and powdered silk fibroin in relatively mild conditions was investigated. A mono-enzymatic hydrolysate systems were tested for hydrolysis of water-soluble form of fibroin as most productive form of protein substrate. Insoluble forms of substrate usually were hydrolyzed less effective. In some cases from soluble fibroin substrate gel was formed during hydrolysis process. This hindered intermixing and decreased rates of hydrolysis. Insoluble sediments were formed in enzymatic hydrolysates in other cases. These sediments and also sediment after chemical hydrolysis were purified and tested on amino acids content for comparison. Sediments formation in these conditions are considered as pure tyrosine isolation method. Obtained hydrolysates were characterized by gel-chromatography analysis and other standard biochemical methods. Possibility of application of enzymatic hydrolysis for preparation of silk fibroin hydrolysates is discussed.