Purification and kinetics of two novel thermophilic extracellular proteases from Lactobacillus helveticus, from kefir with possible biotechnological interest

Valasaki, Krystalenia; Staikou, Aggeliki; Theodorou, Leonidas G.; Charamopoulou, Vasiliki; Zacharaki, Paraskevi; Papamichael, Emmanuel M.

doi:10.1016/j.biortech.2007.10.018

Cited by 21 publications

(11 citation statements)

References 42 publications

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“…Therefore, the proteinase activity detected in our study is mainly from cysteine proteinases. This is also reinforced by the enzyme activity profiles under different pH conditions observed in our study, where both proteinase fractions exhibited two similar peaks of activity-one at acid pH and other at basic pH-suggesting multiple forms of cysteine proteinases in each fraction, both within the range reported for this class of proteinases (Pereira et al 2001;Fahmy et al 2004;Valasaki et al 2008;Silva et al 2010b). However, since cysteine proteinases are not as stable at acidic pH, the peak of activity at lower pH conditions may be due to their autoprocessing at low pH levels.…”

Section: Discussionsupporting

confidence: 77%

“…The temperature profile of activity of the cysteine proteinases obtained from the velvetbean caterpillar in our study also falls within the range reported for cysteine proteinases (Visessanguan et al 2003;Valasaki et al 2008;Silva et al 2010b), which were distinct for both fractions obtained. While the soluble proteinases showed optimum activity at 20 and 35°C, the membrane-bound proteinases showed optimum activity at 60°C, the highest temperature assessed.…”

Section: Discussionsupporting

confidence: 63%

“…In contrast, the aspartate-proteinase inhibitor pepstatin A exhibited negligible effect on both proteinase fractions from the velvetbean caterpillar, as expected (Fahmy et al 2004), which together with the inhibition profiles of E-64 and TLCK provides further support for their recognition as cysteine proteinases. The kinetic parameters determined for the cysteineproteinase fractions of the velvetbean caterpillar are also consistent with those of cysteine proteinases from other species (Visessanguan et al 2003;Fahmy et al 2004;Valasaki et al 2008;Silva et al 2010b), and distinct from those of digestive serine proteinases from the velvetbean caterpillar Xavier et al 2005). The affinity and activity of cysteine proteinases from the velvetbean caterpillar were lower than those observed for the trypsin-like serine proteinases from this same species (lower K m and higher V max values) Xavier et al 2005), indicating the higher catalytic efficiency of the later group of digestive proteinases in the velvetbean caterpillar.…”

Section: Discussionsupporting

confidence: 62%

“…Such trend towards higher cysteine-proteinase activity at higher temperatures (ca. 60°C) was also observed in the lactic-acid producing bacterium Lactobacillus helveticus and in the arrowtooth flounder Atheresthes stomias (Visessanguan et al 2003;Valasaki et al 2008).…”

Section: Discussionmentioning

confidence: 71%

“…. Calcium-dependent and independent forms of cysteine proteinases have been reported (Valasaki et al 2008;Silva et al 2010b).…”

Section: Discussionmentioning

confidence: 99%

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Midgut cysteine-proteinase activity in the velvetbean caterpillar (Anticarsia gemmatalis (Hübner))

et al. 2011

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Proteinase inhibitors are currently targeted as potential insect control agents, but adaptation to proteinase inhibitors is a recognized limitation to such approach requiring the understanding of how phytophagous species can cope with such compounds. The velvetbean caterpillar (Anticarsia gemmatalis) is a key pest of soybean and welladapted to its host proteinase inhibitors, which is rich in serine-proteinase inhibitors, particularly trypsin-like proteinase inhibitors. As the expression of cysteine proteinases in the midgut of the velvetbean caterpillar is a potential adaptation to circumvent its host defense, we assessed and characterized the digestive cysteine-proteinase activity from velvetbean caterpillars. Significant soluble and membranebound proteolytic activity was obtained and was consistent with those of cysteine proteinases based on the substrate and inhibitors used for their characterization. The K m and V max values obtained were 2.35 ± 0.50 mM and 40.89 ± 6.68 nmol min -1 mg -1 for the soluble proteinases and 0.33 ± 0.03 mM and 24.54 ± 0.67 nmol min -1 mg -1 for the membrane-bound proteinases, range of values also consistent with cysteine proteinases. Therefore, the proteolytic activity observed in the velvetbean caterpillar midgut is consistent with that of cysteine proteinases providing preliminary support for the contention of their potential involvement mitigating the negative effects of serineprotease inhibitors in this species.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionsupporting

confidence: 63%

Section: Discussionsupporting

confidence: 62%

Section: Discussionmentioning

confidence: 71%

“…. Calcium-dependent and independent forms of cysteine proteinases have been reported (Valasaki et al 2008;Silva et al 2010b).…”

Section: Discussionmentioning

confidence: 99%

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Midgut cysteine-proteinase activity in the velvetbean caterpillar (Anticarsia gemmatalis (Hübner))

et al. 2011

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Synthesis of palladium nanoparticles utilizing biotemplates and investigation of their synergistic catalytic performance

Xie,

Chen,

2024

J of Chemical Tech & Biotech

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BACKGROUNDPalladium nanoparticles can act as a shuttle to accelerate the extracellular electron transfer (EET) by exoelectrogens. Through the EET process, microorganisms drive the redox cycle of many substances. The palladium nanoparticle has a variety of catalytic activities and activities can be significantly improved by combining with the active effects of microorganisms.RESULTSIn this study, Staphylococcus saprophyticus JJ‐1 was utilized as a biological template carrier to synthesize palladium nanoparticles that were immobilized on the bacterial surface. The morphology and composition of palladium were characterized by various techniques including transmission electron microscopy, energy‐dispersive X‐ray spectroscopy, X‐ray diffraction, X‐ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy. Furthermore, the hydrogenation activity of the synthesized Pd was tested at room temperature and atmospheric pressure using methyl orange as a model pollutant. Electrochemical characterization was carried out by cyclic voltammetry and potentiostatic measurements. The research demonstrated that palladium nanoparticles acted as both electronic mediators and catalysts in the dye reduction process, while S. saprophyticus JJ‐1 contributed to stabilizing nanoparticles and electrochemical activity. The synergistic effect between these two components significantly enhances MO degradation efficiency.CONCLUSIONThis study presented an energy‐saving method to synthesize an integrated catalyst based on the synergistic interaction between biomass and nanoparticles, offering a novel approach for developing environmentally friendly, cost‐effective, and efficient integrated catalysts. © 2024 Society of Chemical Industry (SCI).

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Proteolytic activity of gut bacteria isolated from the velvet bean caterpillar Anticarsia gemmatalis

et al. 2013

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The development of proteinase inhibitors as potential insect control agents has been constrained by insect adaptation to these compounds. The velvet bean caterpillar (Anticarsia gemmatalis) is a key soybean pest species that is well-adapted to proteinase inhibitors, particularly serine-proteinase inhibitors, which are abundant in the caterpillar host. The expression of diverse proteolytic enzymes by gut symbionts may allow the velvet bean caterpillar to circumvent proteinase inhibitors produced by the host plant. In this study, we characterized the proteolytic activity of the four nonpathogenic species of gut bacteria isolated from the velvet bean caterpillar-Bacillus cereus, Enterococcus gallinarum, Enterococcus mundtii and Staphylococcus xylosus. Two proteinase substrates, N-α-benzoyl-L-Arg-p-nitroanilide (L-BApNA) and N-α-p-tosyl-L-Arg methyl ester (L-TAME) and five proteinase inhibitors [aprotinin, E-64, ethylenediamine tetraacetic acid (EDTA), pepstatin and N-α-tosyl-L-lysine chloromethyl ketone (TLCK)] as well as CaCl2, pH and temperature profiles were used to characterize the expressed proteolytic activity of these bacterial strains in vitro. Kinetic parameters for proteolytic activity were also estimated. The results of these experiments indicated that serine- and cysteine-proteinase activities were expressed by all four gut bacteria symbionts of the velvet bean caterpillar. The cysteine- and serine-proteinase activities of these gut symbionts were distinct and different from that of gut proteinases of the caterpillar itself. This finding provides support for the potential involvement of gut symbionts in the mitigation of the negative effects of serine-proteinase inhibitors in the velvet bean caterpillar.

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Purification and kinetics of two novel thermophilic extracellular proteases from Lactobacillus helveticus, from kefir with possible biotechnological interest

Cited by 21 publications

References 42 publications

Midgut cysteine-proteinase activity in the velvetbean caterpillar (Anticarsia gemmatalis (Hübner))

Midgut cysteine-proteinase activity in the velvetbean caterpillar (Anticarsia gemmatalis (Hübner))

Synthesis of palladium nanoparticles utilizing biotemplates and investigation of their synergistic catalytic performance

Proteolytic activity of gut bacteria isolated from the velvet bean caterpillar Anticarsia gemmatalis

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