Purification and properties of serine protease from Halobacterium halobium

Izotova, Lara S.; Strongin, Alex Y.; Chekulaeva, L. N.; Sterkin, V E; Ostoslavskaya, V. I.; Lyublinskaya, L. A.; Тимохина, Е. А.; Stepanov, V. M.

doi:10.1128/jb.155.2.826-830.1983

Cited by 96 publications

(34 citation statements)

References 17 publications

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“…Concerning the protease producers, they represent 17% of the hydrolytic population. Several proteolytic enzymes have been isolated and characterized from Archaea (Izotova et al, 1983;Kamekura & Seno, 1990;Yu, 1991;Stepanov et al, 1992;Ryu et al, 1994;Giménez et al, 2000;Studdert et al, 2001); however, no protease from extremely halophilic bacteria has been described so far.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Moreno

Garcı́a²,

Ventosa

et al. 2009

FEMS Microbiology Ecology

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In order to explore the diversity of extreme halophiles able to produce different hydrolytic enzymes (amylase, protease, lipase and DNAse) in hypersaline habitats of South Spain, a screening program was performed. A total of 43 extreme halophiles showing hydrolytic activities have been isolated and characterized. The isolated strains were able to grow optimally in media with 15-20% (w/v) total salts and in most cases, growth was detected up to 30% (w/v) total salts. Most hydrolase producers were assigned to the family Halobacteriaceae, belonging to the genera Halorubrum (22 strains), Haloarcula (nine strains) and Halobacterium (nine strains), and three isolates were characterized as extremely halophilic bacteria (genera Salicola, Salinibacter and Pseudomonas). An extremely halophilic isolate, strain IC10, showing lipase and protease activities and identified as a Salicola strain of potential biotechnological interest, was further studied. The optimum growth conditions for this strain were 15-20% (w/v) NaCl, pH 8.0, and 37 degrees C. Zymographic analysis of strain IC10 detected the lipolytic activity in the intracellular fraction, showing the highest activity against p-nitrophenyl-butyrate as a substrate in a colorimetric assay, whereas the proteolytic activity was detected in the extracellular fraction. This protease degraded casein, gelatin, bovine serum albumin and egg albumin.

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Section: Discussionmentioning

confidence: 99%

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Moreno

Garcı́a²,

Ventosa

et al. 2009

FEMS Microbiology Ecology

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“…Although many of the above applications have traditionally relied on proteases of animal and plant origin, microbial proteases have become more widely acceptable and available. Many proteases have been purified and characterized from bacteria (Izotova et al 1983;Monmaney 1985;Durham et al 1987), fungi (Klapper et al 1973;Dworschack 1952) and yeasts (Maddox and Hough 1970;Alessandro and Federico 1980;Nelson and Young 1986).…”

Section: Introductionmentioning

confidence: 99%

Production and properties of an alkaline protease by Aureobasidium pullulans

Donaghy¹,

McKay

1993

Journal of Applied Bacteriology

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A strain of the yeast‐like fungus Aureobasidium pullulans was grown on whey to produce an extracellular protease. The protease was totally inhibited by the serine inhibitor, phenyl methyl sulphonyl fluoride (PMSF), and partially inhibited by the chelating agent EDTA. The enzyme showed maximal activity in the alkaline range with an optimum pH of 9·5–10·5. The optimum temperature for protease activity was 41d̀C. As well as being active against the non‐specific proteolytic substrate Azocoll, the protease readily degraded purified α‐casein. A molecular weight of 27000 ± 350 was determined for the protease using gel filtration chromatography.

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“…The addition of aspartic acid, glutamic acid and glycine did not result in the maximal protease production. The addition of these amino acids resulted in the marked increase in protease production of some halophilic bacteria (Izotova et al 1983;Stepanov et al 1992;Namwong et al 2005Namwong et al , 2006. As halophilic proteases generally have remarkably high contents of glycine, aspartic acid and glutamic acid, these amino acids may enable cells to evolve proteins that are exceedingly rich in acidic amino acids.…”

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confidence: 99%

CHARACTERIZATION OF HALOPHILIC BACTERIUM NB2-1 FROMPLA-RAAND ITS PROTEASE PRODUCTION

Chamroensaksri

Akaracharanya

Visessanguan

et al. 2008

Journal of Food Biochemistry

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A halophilic proteolytic bacterium, NB2‐1, isolated from Pla‐ra, fermented fish in Thailand, was identified as Virgibacillus marismortui on the basis of the genetic and phenotypic characteristics. NB2‐1 produced a substantial level of extracellular proteolytic activity, corresponding with the growth and reaching the maximum at the end of the exponential phase (3 days). The optimum conditions required for maximum protease production were 15% (w/v) NaCl, pH at 9 and temperature at 30C. Of the media tested, maximum protease production occurred in a medium containing 15% NaCl, 0.5% yeast extract, 0.1% glutamic acid, 0.2% KCl, 0.3% trisodium citrate, 2% MgSO4·7H2O, 0.0036% FeCl2·4H2O and 0.000036% MnCl2·4H2O. The protease activity was optimally active at pH 10, 50C and 5% (w/v) NaCl when casein was used as a substrate. The activity was strongly inhibited by phenylmethane sulfonyl fluoride. Activity‐stained substrate gel indicated the presence of several proteases with the estimated molecular weights of 17, 19, 24, 29 and 35 kDa in the crude NB2‐1 extracellular protease. The results of the present investigation showed that the strain NB2‐1 could be a potential source of thermoactive alkaline protease production. PRACTICAL APPLICATIONS Halophiles are the most likely source of enzymes that not only are salt‐tolerant but also thermotolerant. They constitute a heterogeneous group of microorganisms including species belonging to different genera. The isolation of halophiles able to produce extracellular enzymes will provide the possibility to have optimal activities at different salt concentrations. Virgibacillus marismortui NB2‐1, a moderately halophilic bacterium isolated from Pla‐ra, produced a substantial level of extracellular proteolytic activity that was active in extreme conditions. Using casein as a substrate, the protease exhibited maximal activity at pH 10, 50C in the presence of 5% (w/v) NaCl. The results of the present investigation showed that the strain NB2‐1 could be a potential source of thermoactive alkaline protease production.

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Purification and properties of serine protease from Halobacterium halobium

Cited by 96 publications

References 17 publications

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Production and properties of an alkaline protease by Aureobasidium pullulans

CHARACTERIZATION OF HALOPHILIC BACTERIUM NB2-1 FROMPLA-RAAND ITS PROTEASE PRODUCTION

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Purification and properties of serine protease from Halobacterium halobium

Cited by 96 publications

References 17 publications

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Production and properties of an alkaline protease by Aureobasidium pullulans

CHARACTERIZATION OF HALOPHILIC BACTERIUM NB2-1 FROMPLA-RAAND ITS PROTEASE PRODUCTION

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Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile

Characterization of Salicola sp. âIC10, a lipase- and protease-producing extreme halophile