Antibacterial serine protease from Wrightia tinctoria : Purification and characterization

Muthu, Sakthivel; Gopal, Venkatesh Babu; Soundararajan, Selvakumar; Karthikeyan, N.; Narayan, Karthik S.; Lakshmikanthan, Mythileeswari; Sathuvan, Malairaj; Palani, P.

doi:10.1016/j.plaphy.2017.01.003

Cited by 15 publications

(8 citation statements)

References 47 publications

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“…Results close to the study were verified in the work of Muthu et al (2017) for proteins quantification (0.23 mg/mL to 1.55 mg/mL) in extracts from 20 Factors such as temperature, pH and the type of protease present in crude vegetable extracts affect the determination of milk clotting activity (Mazorra-Manzano et al, 2013) and consequently its potential for replacing rennet. The results of milk clotting activity (MCA) and rennet substitution potential (RSP) of the artichoke (Cynara scolymus) leaves extracts are shown in Table 2.…”

Section: Coagulation Activity and Rennet Substitution Potential Of Artichoke Leaf Extractssupporting

confidence: 64%

Artichoke leaf extracts: Proteolytic activity, coagulant and HPLC analysis

et al. 2021

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The search for origin plant-based proteases increases gradually due to their diversity and stability over a wide range of pH and temperature. Artichoke (Cynara scolymus) flowers are a proteolytic vegetable source already studied, but their leaves are scarce in this respect. Thus, the objective of this research was to obtain extracts of artichoke leaves with different buffers and extraction methods as an alternative proteolytic source and plant coagulant, as well as the separation and comparison of the protein profile of these extracts. The methodology used was based on extraction with sodium citrate buffer (pH 3), sodium acetate (pH 5) and Tris-HCl (pH 7) by mechanical stirrer (MS) and ultrasound (US); protein determination; proteolytic activity (PA) and specific activity (SA); milk clotting activity (MCA) and rennet substitution potential (RSP); high- performance liquid chromatography analysis (HPLC) with UV-Vis detector and principal component analysis (PCA). Extracts of Cynara scolymus leaves showed high results with Citrate-US for the parameters PA (14.38), SA (19.71), MCA (440) and RSP (30.60) compared to other treatments. The extracts with citrate and acetate presented a quick coagulation time (max 3 min). The HPLC analysis enabled the separation of the different protein compounds present in the extracts and most expressive peaks in the samples with Citrate-MS and Acetate-MS; and isolated peaks for Citrate-US. It was concluded that extracts of artichoke leaves with citrate and acetate buffer attributed satisfactory results to act as plant coagulant, as well as to carry out further studies for the purification of proteolytic enzymes and application in cheeses.

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Section: Coagulation Activity and Rennet Substitution Potential Of Artichoke Leaf Extractssupporting

confidence: 64%

Artichoke leaf extracts: Proteolytic activity, coagulant and HPLC analysis

et al. 2021

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“…; Muthu et al . ). With the purpose of corroborating if salpichroin was also capable of inhibiting bacterial growth, we ascertained its activity against two human pathogens, Escherichia coli and Staphylococcus aureus .…”

Section: Resultsmentioning

confidence: 97%

Antimicrobial activity of an aspartic protease fromSalpichroa origanifoliafruits

Díaz

Rocha

Kise

et al. 2018

Lett Appl Microbiol

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This study provides insights into the antimicrobial activity of an aspartic protease isolated from Salpichroa origanifolia fruits on plant and human pathogens. The proteinase inhibited Fusarium solani and Staphylococcus aureus in a dose-dependent manner due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of salpichroin suggests its potential applications as an important tool for the control of pathogenic micro-organisms affecting humans and crops of economic interest. Therefore, it would represent a new alternative to avoid the problems of environmental pollution and antimicrobial resistance.

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“…Numerous independent studies confirm that serine proteases hold promise for medical purposes (Yariswamy et al, 2013;Muthu et al, 2017;Belov et al, 2018;Abaturov, 2020;Osmolovsky et al, 2020). In particular, one of the main problems that physicians face when treating skin wounds and burns in people with compromised immunity is the formation of a surface biofilm generated by conditionally pathogenic microorganisms (Staphylococcus aureus and S. epidermidis, micrococci, and Pseudomonas); under this biofilm, the microbes cannot be reached by antibiotics, and as a consequence, wound healing slows down.…”

Section: Introductionmentioning

confidence: 99%

Creation of a recombinant Komagataella phaffii strain, a producer of proteinase K from Tritirachium album

et al. 2022

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The objects of the study were recombinant clones of Komagataella phaffii K51 carrying the heterologous proteinase K (PK-w) gene from Tritirachium album integrated into their genome as well as samples of recombinant proteinase K isolated from these clones. The aims of this work were i) to determine whether it is possible to create recombinant K. phaffii K51 clones overexpressing functionally active proteinase K from T. album and ii) to analyze the enzymatic activity of the resulting recombinant enzyme. The following methods were used: computational analysis of primary structure of the proteinase K gene, molecular biological methods (PCR, electrophoresis of DNA in an agarose gel, electrophoresis of proteins in an SDS polyacrylamide gel under denaturing conditions, spectrophotometry, and quantitative assays of protease activity), and genetic engineering techniques (cloning and selection of genes in bacterial cells Escherichia coli TOP10 and in the methylotrophic yeast K. phaffii K51). The gene encoding natural proteinase K (PK-w) was designed and optimized for expression in K. phaffii K51. The proteinase K gene was synthesized and cloned within the plasmid pPICZα-A vector in E. coli TOP10 cells. The proteinase K gene was inserted into pPICZα-A in such a way that – at a subsequent stage of transfection into yeast cells – it was efficiently expressed under the control of the promoter and terminator of the AOX1 gene, and the product of the exogenous gene contained the signal peptide of the Saccharomyces cerevisiae a-factor to ensure the protein’s secretion into the culture medium. The resultant recombinant plasmid (pPICZα-A/PK-w) was transfected into K. phaffii K51 cells. A recombinant K. phaffii K51 clone was obtained that carried the synthetic proteinase K gene and ensured its effective expression and secretion into the culture medium. An approximate productivity of the yeast recombinant clones for recombinant proteinase K was 25 μg/ mL after 4 days of cultivation. The resulting recombinant protease has a high specific proteolytic activity: ~5000 U/mg.

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Antibacterial serine protease from Wrightia tinctoria : Purification and characterization

Cited by 15 publications

References 47 publications

Artichoke leaf extracts: Proteolytic activity, coagulant and HPLC analysis

Artichoke leaf extracts: Proteolytic activity, coagulant and HPLC analysis

Antimicrobial activity of an aspartic protease fromSalpichroa origanifoliafruits

Creation of a recombinant Komagataella phaffii strain, a producer of proteinase K from Tritirachium album

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