Producing marker-free Kalanchoe plants expressing antimicrobial peptide cecropin P1 gene

Захарченко, Н. С.; Лебедева, А. А.; Furs, O. V.; Рукавцова, Е. Б.; Schevchuk, T. V.; Rodionov, I. L.; Buryanov, Yа. I.

doi:10.1134/s1021443716020163

Cited by 5 publications

(5 citation statements)

References 27 publications

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“…Presence of CecP1 in the extracts was tested using immunoblotting as described formerly [ 10 ] ( Figure 2 ). Briefly, 10.0 μ L of extract and 30.0 μ L of the control synthetic CecP1 (1 μ g/mL) were pelleted with 10% trichloroacetic acid, denatured and separated in 15% polyacrylamide gel, blotted to nitrocellulose membrane, and stained with rabbit polyclonal antibodies.…”

Section: Resultsmentioning

confidence: 99%

“…Sophisticated methods of its producing by recombinant strains within fused proteins with decreased toxicity are expensive and difficult for technological implementation [ 9 ]. However, there is a broad range of data about microbicide activity of CecP1 towards bacterial and fungal phytopathogens [ 10 ] and human/animal pathogens [ 11 ]. Virucide and antitumor activity was also attributed to CecP1 [ 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…Recently engineering transgenic plants of Kalanchoe pinnata expressing CecP1 synthetic gene and accumulating the recombinant AMP in cytoplasm was reported [ 10 ]. Bactericide efficiency of pure CecP1 and the wild-type K. pinnata extract against model bacterial strains is described [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

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Bactericide, Immunomodulating, and Wound Healing Properties of TransgenicKalanchoe pinnataSynergize with Antimicrobial Peptide Cecropin P1 In Vivo

Лебедева

Захарченко

Трубникова

et al. 2017

Journal of Immunology Research

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Procedure of manufacturing K. pinnata water extracts containing cecropin P1 (CecP1) from the formerly described transgenic plants is established. It included incubation of leaves at +4°C for 7 days, mechanical homogenization of leaves using water as extraction solvent, and heating at +70°C for inactivating plant enzymes. Yield of CecP1 (after heating and sterilizing filtration) was 0.3% of total protein in the extract. The water extract of K. pinnata + CecP1 exhibits favorable effect on healing of wounds infected with S. aureus (equal to Cefazolin) and with a combination of S. aureus with P. aeruginosa (better than Cefazolin). Wild-type K. pinnata extract exhibited evident microbicide activity against S. aureus with P. aeruginosa but it was substantially strengthened in K. pinnata + CecP1 extract. K. pinnata extracts (both wild-type and transgenic) did not exhibit general toxicity and accelerated wound recovery. Due to immunomodulating activity, wild-type K. pinnata extract accelerated granulation of the wound bed and marginal epithelialization even better than K. pinnata + CecP1 extract. Immunomodulating and microbicide activity of K. pinnata synergizes with microbicide activity of CecP1 accelerating elimination of bacteria.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Bactericide, Immunomodulating, and Wound Healing Properties of TransgenicKalanchoe pinnataSynergize with Antimicrobial Peptide Cecropin P1 In Vivo

Лебедева

Захарченко

Трубникова

et al. 2017

Journal of Immunology Research

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“…Recently, transgenic plants of K. pinnata producing antimicrobial peptide (AMP) cecropin P1 (CecP1) from Ascaris suis was described [ 27 ]. An antimicrobial, antifungal, and antiviral activity was attributed to this AMP [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Immunomodulating and Revascularizing Activity of Kalanchoe pinnata Synergize with Fungicide Activity of Biogenic Peptide Cecropin P1

Захарченко

Белоус

Бирюкова

et al. 2017

Journal of Immunology Research

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Previously transgenic Kalanchoe pinnata plants producing an antimicrobial peptide cecropin P1 (CecP1) have been reported. Now we report biological testing K. pinnata extracts containing CecP1 as a candidate drug for treatment of wounds infected with Candida albicans. The drug constitutes the whole juice from K. pinnata leaves (not ethanol extract) sterilized with nanofiltration. A microbicide activity of CecP1 against an animal fungal pathogen in vivo was demonstrated for the first time. However, a favorable therapeutic effect of the transgenic K. pinnata extract was attributed to a synergism between the fungicide activity of CecP1 and wound healing (antiscar), revascularizing, and immunomodulating effect of natural biologically active components of K. pinnata. A commercial fungicide preparation clotrimazole eliminated C. albicans cells within infected wounds in rats with efficiency comparable to CecP1-enriched K. pinnata extract. But in contrast to K. pinnata extract, clotrimazole did not exhibit neither wound healing activity nor remodeling of the scar matrix. Taken together, our results allow assumption that CecP1-enriched K. pinnata extracts should be considered as a candidate drug for treatment of dermatomycoses, wounds infected with fungi, and bedsores.

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“…Because of its widespread distribution and ubiquitous ethnobotanical use, much research has been focused on K. pinnata , a species native to Madagascar but cultivated and distributed throughout the tropics (Descoings, 2003; Biswas et al, 2011a; Quazi Majaz et al, 2011; Pattewar, 2012; Rajsekhar et al, 2016). This species has even been the subject of bioengineering – a transgenic K. pinnata that produces an antimicrobial peptide (AMP cecropin P1) has recently been developed (Zakharchenko et al, 2016; Lebedeva et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Antibacterial Activity of Kalanchoe mortagei and K. fedtschenkoi Against ESKAPE Pathogens

et al. 2019

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Plants in the genus Kalanchoe (Family: Crassulaceae) are used in traditional medicine throughout the tropics for treating a variety of conditions. Two species, Kalanchoe mortagei and K. fedtschenkoi, have established ethnobotanical usage but have been neglected in previous research concerning their potential bioactivity. Here, we provide a thorough review of the reported antimicrobial activities of Kalanchoe genus and evaluate the in vitro antibacterial effects of two previously unexplored species against a panel of multidrug-resistant bacteria, the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter cloacae). Plant specimens were collected and voucher specimens deposited in the Emory University Herbarium. Dried plant material was ground into a powder and extracted as ethanolic macerations or as aqueous decoctions. Extracts were tested against the ESKAPE pathogens for growth inhibitory activity. Cytotoxicity to human cells was assessed via a lactate dehydrogenase assay of treated human keratinocytes (HaCaTs). K. fedtschenkoi extracts demonstrated growth inhibitory effects against two Gram-negative species, A. baumannii (strain CDC-33) and P. aeruginosa (AH-71), as well as S. aureus (UAMS-1). In these cases, growth inhibition greater than 50% (IC50) was generally observed at concentrations of 256 μg mL-1, though one K. fedtschenkoi extract (1465, prepared from stems) exhibited an IC50 against A. baumannii at 128 μg mL-1. All extracts were well tolerated by HaCaTs (LD50 ≥ 256 μg mL-1). Chemical characterization using HPLC and chemical standards established the presence of caffeic acid and quercetin in both plant species, as well as kaempferol in K. fedtschenkoi. These results reveal K. fedtschenkoi to be a plant of medicinal interest, and future research should aim to characterize the bioactivity of this species and its active constituents through bioassay-guide fractionation. Effects on bacterial biofilm formation and quorum-sensing are also research topics of interest for this genus.

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Producing marker-free Kalanchoe plants expressing antimicrobial peptide cecropin P1 gene

Cited by 5 publications

References 27 publications

Bactericide, Immunomodulating, and Wound Healing Properties of TransgenicKalanchoe pinnataSynergize with Antimicrobial Peptide Cecropin P1 In Vivo

Bactericide, Immunomodulating, and Wound Healing Properties of TransgenicKalanchoe pinnataSynergize with Antimicrobial Peptide Cecropin P1 In Vivo

Immunomodulating and Revascularizing Activity of Kalanchoe pinnata Synergize with Fungicide Activity of Biogenic Peptide Cecropin P1

Antibacterial Activity of Kalanchoe mortagei and K. fedtschenkoi Against ESKAPE Pathogens

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