Purification and characterization of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp.

Patel, Rajesh; Dodia, M. S.; Joshi, Rupal H.; Singh, Satya P.

doi:10.1016/j.procbio.2006.04.016

Cited by 82 publications

(48 citation statements)

References 40 publications

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“…These results were similar to those of thermostable alkaline protease from Bacillus sp. (Patil et al, 2010) and PLLA-degrading serine protease from Actinomadura sp. T16-1 (Sukkhum et al, 2009b).…”

Section: Effect Of Detergents Inhibitors and Metal Ions Of Purified mentioning

confidence: 99%

Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175

Hanphakphoom

Maneewong

Sukkhum

et al. 2014

J. Gen. Appl. Microbiol.

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on an emulsified PLLA agar plate at 50 C. Among the isolates, strain LP175 showed the highest PLLAdegrading ability. It was closely related to Laceyella sacchari, with 99.9% similarity based on the 16S rRNA gene sequence. The PLLA-degrading enzyme produced by the strain was purified to homogeneity by 48.1% yield and specific activity of 328 U·mg-protein-1 with a 15.3-fold purity increase. The purified enzyme was strongly active against specific substrates such as casein and gelatin and weakly active against Suc-(Ala) 3 -pNA. Optimum enzyme activity was exhibited at a temperature of 60 C with thermal stability up to 50 C and a pH of 9.0 with pH stability in a range of 8.5 10.5. Molecular weight of the enzyme was approximately 28.0 kDa, as determined by gel filtration and SDS-PAGE. The inhibitors phenylmethylsulfonyl fluoride (PMSF), ethylenediaminetetraacetate (EDTA), and ethylene glycolbis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) strongly inhibited enzyme activity, but the activity was not inhibited by 1 mM 1,10-phenanthroline (1,10-phen). The N-terminal amino acid sequences had 100% homology with thermostable serine protease (thermitase) from Thermoactinomyces vulgaris. The results obtained suggest that the PLLA-degrading enzyme produced by L. sacchari strain LP175 is serine protease.Key words: Laceyella sacchari; PLLA-degrading enzyme; Thermoactinomycetaceae; thermophilic filamentous bacteria IntroductionPoly(L-lactide) (PLLA); (C 3 H 4 O 2 )n is one of the aliphatic biodegradable polyesters derived from renewable resources such as corn, cassava, sugar cane, rice and potato through lactic acid fermentation, and it is also fully degradable by both microbial and enzymatic processes (Tokiwa et al., 2009;Gupta et al., 2007;Jarerat et al., 2006;Tokiwa and Calabia, 2006;Tomita et al., 2004). PLLA, as an environmentally friendly (eco-friendly or green ) product, has been developed on a large scale and is currently used for a wide range of applications, including packaging materials (Bhalla et al., 2007;Nolan-Itu Pty Ltd., 2002), medical applications (Jalil, 1990), agricultural products (Gross and Kalra, 2002;Sakai et al., 2001) and textiles.Most PLLA-degrading microorganisms are found in the bacterial order Actinomycetales, e.g. the families Pseudonocardiaceae (Pranamuda and Tokiwa, 1999;Pranamuda et al., 1997), Thermomonosporaceae (Sangwan and Wu, 2008;Sukkhum et al., 2009b), and Streptosporangiaceae (Sukkhum et al., 2009b). Apart from that, thermophilic bacteria in the families Thermoactinomycetaceae (Sukkhum et al., 2009b) and Firmicutes (Oda et al., 2000;Sakai et al., 2001), filamentous fungi in the genera Tritirachium and Paecilomyces (Sangwan and Wu, 2008), and yeast in the genus Cryptococcus have also been reported to be PLLA-degrading microorganisms.Many different types of enzymes are found in PLLAdegrading microorganisms, such as: protease from Amycola- Full PaperCharacterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175 (Received F...

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Section: Effect Of Detergents Inhibitors and Metal Ions Of Purified mentioning

confidence: 99%

Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175

Hanphakphoom

Maneewong

Sukkhum

et al. 2014

J. Gen. Appl. Microbiol.

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“…Salt tolerant proteases are highly specific in their action and their activity can be better standardized in any harsh environment. Several researchers reported the novel characteristics of salt tolerant/resistant proteases identified from different microbes (4,10,11,12,13,17,20,21,22). Such kind of enzyme characteristics encourages their application in wastewater treatment systems.…”

Section: Introductionmentioning

confidence: 99%

Optimization studies on production of a salt-tolerant protease from Pseudomonas aeruginosa strain BC1 and its application on tannery saline wastewater treatment

Sivaprakasam¹,

Dhandapani²,

Surianarayanan³

2011

Braz. J. Microbiol.

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Treatment and safe disposal of tannery saline wastewater, a primary effluent stream that is generated by soaking salt-laden hides and skin is one of the major problems faced by the leather manufacturing industries.Conventional treatment methods like solar evaporation ponds and land composting are not eco-friendly as they deteriorate the ground water quality. Though, this waste stream is comprised of high concentration of dissolved proteins the presence of high salinity (1-6 % NaCl by wt) makes it non-biodegradable. Enzymatic treatment is one of the positive alternatives for management of such kind of waste streams. A novel salttolerant alkaline protease obtained from P.aeruginosa (isolated from tannery saline wastewater) was used for enzymatic degradation studies. The effect of various physical factors including pH, temperature, incubation time, protein source and salinity on the activity of identified protease were investigated. Kinetic parameters (K m , V max ) were calculated for the identified alkaline protease at varying substrate concentrations. Tannery saline wastewater treated with identified salt tolerant protease showed 75 % protein removal at 6 h duration and 2 % (v/v) protease addition was found to be the optimum dosage value.

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“…was found active in the range of pH 8.5-12.0 with the optima at 10.0-11.0. The requirement of salt for enzyme catalysis increased with increase in temperature, and a shift in temperature optima from 37 o C to 55 o C was evident in the presence of 2% salt [173]. The protease from haloalkaliphilic archaeon Natrialba magadii required high salt concentration to exhibit elevated stability in the presence of organic solvents [174].…”

Section: Alkaline Proteases From Halophilesαmentioning

confidence: 99%

Extreme Environments: Goldmine of Industrially Valuable Alkali-stable Proteases

Garg¹,

Singh²

2015

Enz Eng

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Since the advent and release of Bio 40 (the first detergent with bacterial alkaline protease) in 1959, exploration of the microbial alkaline proteases has been exploited beyond expectations. The inventory of microbial alkaline proteases is difficult to draft as it is increasing daily. Most of these proteases are the result of studies in which one or a few isolates were targeted for an alkali-stable enzyme. Although, the worldwide research on microbial alkaline proteases has been widely performed, we still need an improved enzyme capable of catalyzing reactions under extreme conditions (also called as extremozymes), e.g., high alkalinity and salinity, anhydrous environment, cold and hot environment, etc. As the search for extremozymes is in progress, it is imperative to explore the extreme environments to get some novel microbial strains (extremophiles) for alkali-stable proteases. This review article is an effort to compile the scattered information on some extremophiles and their alkali-stable proteases, which may have better specific industrial applications. It includes: (i) various extreme environments relevant to industrial biocatalysts, (ii) strategies of extremophilic microbes and enzymes which make them able to tolerate such conditions, and (iii) an overview of some important work done so far to explore industrially relevant extremophilic enzymes from extremophiles.

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Purification and characterization of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp.

Cited by 82 publications

References 40 publications

Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175

Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175

Optimization studies on production of a salt-tolerant protease from Pseudomonas aeruginosa strain BC1 and its application on tannery saline wastewater treatment

Extreme Environments: Goldmine of Industrially Valuable Alkali-stable Proteases

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