scholarly journals Purification, characterization, and bioassay of putative protease inhibitors from Hevea brasiliensis latex

2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Riza Arief PUTRANTO ◽  
. SISWANTO ◽  
Agustin Sri MULYATNI ◽  
Asmini BUDIANI ◽  
Radite TISTAMA
Keyword(s):  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Inhibitory Activity ◽  
Hevea Brasiliensis ◽  
Rubber Tree ◽  
Exchange Chromatography ◽  
Mycelium Growth ◽  
Plant Protease ◽  
Putative Protease

Latex, a milky white liquid, is the main product from rubber tree (Hevea brasiliensis). Latex is the cytoplasm of complex cellular networks named laticifers in which it contains many different components, including important proteins. Various types of enzymes carrying functions associated with plant defense against pathogen and wounding have been detected in latex in which one of these enzymes is protease inhibitor (PI). Plant protease inhibitor has tremendous potential as an antifungal agent which can be developed as biofungicide. In this work, protease inhibitors from B-serum (lutoid) of rubber tree latex were isolated and purified using Ion Exchange Chromatography (IEC) technique. Of the total 70 fractions of proteins extracted from the columns, only 26 fractions showed measurable levels of protein. The concentration of obtained putative protease inhibitors (three fractions of IEC) ranged from 0.007 to 0.022 mL/g B-serum. Inhibitory activity against four protease enzymes (subtilisin A, trypsin, α-chymotrypsin, and papain) showed the characteristics of Hevea putative protease inhibitors from B-serum as serine and/or cysteine protease inhibitors with more than 15% inhibitory activity of target protease. Based on SDS-PAGE visualization, the molecular weight of dominant protein considered as Hevea putative protease inhibitors was 21.5 kDa. In vitro bioassay test of antifungal activity for Hevea putative protease inhibitors showed reduced mycelium growth of Ganoderma boninense, Sclerotium sp., and Rigidosporus lignosus.

scholarly journals Purification, characterization, and bioassay of putative protease inhibitors from Hevea brasiliensis latex

2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Riza Arief PUTRANTO ◽  
. SISWANTO ◽  
Agustin Sri MULYATNI ◽  
Asmini BUDIANI ◽  
Radite TISTAMA
Keyword(s):  
Ion Exchange ◽  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Hevea Brasiliensis ◽  
Rubber Tree ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sds Page ◽  
Putative Protease

Lateks yang menyerupai cairan susu putih diperoleh dari penyadapan kulit batang tanaman karet (Hevea brasiliensis). Lateks merupakan sitoplasma dari jaringan pembuluh bernama latisifer yang didalamnya terkandung berbagai macam komponen, termasuk protein-protein penting. Berbagai jenis enzim yang memiliki fungsi terkait pertahanan tanaman dari serangan patogen dan pelukaan telah berhasil dideteksi di dalam lateks, di antaranya protease inhibitor (PI). Protease inhibitor memiliki aktivitas senyawa antifungi sehingga berpotensi untuk  dimanfaatkan sebagai biofungisida. Pada penelitian ini, protease  inhibitor putatif yang berasal dari serum B (lutoid) lateks tanaman karet telah berhasil diisolasi menggunakan teknik Ion Exchange Chroma-tography. Dari total 70 fraksi protein yang diekstrak dari kolom, hanya 26 fraksi yang menunjukkan kadar protein yang terukur. Kandungan protease inhibitor putatif yang di-peroleh berkisar antara 0,0067 hingga 0,022 mL/g serum B dari hasil 3 fraksi terpilih. Aktivitas penghambatan terhadap empat enzim protease (subtilisin A, tripsin, α-kimotripsin, dan papain) menunjukkan karakteristik protease inhibitor putatif tersebut sebagai serine dan/atau cysteine inhibitor protease dengan persentase hambatan di atas 15% terhadap protease target. Hasil SDS-PAGE memperlihatkan pemisahan protein dominan yang diperkirakan merupakan protease inhibitor putatif dengan berat molekul sebesar 21,5 kDa. Uji bioassay aktivitas antifungi secara in vitro dari protease inhibitor memperlihatkan penghambatan pertumbuhan miselium dari fungi Ganoderma boninense, Sclerotium sp., dan Rigidosporus lignosus. [Kata kunci : protease inhibitor, Hevea brasiliensis, lateks, serum B, ion exchange chromatography]AbstractLatex, a milky white liquid, is the main product from rubber tree (Hevea brasiliensis). Latex is the cytoplasm of complex cellular networks named laticifers in which it contains many different components, including important proteins. Various types of enzymes carrying functions associated with plant defense against pathogen and wounding have been detected in latex in which one of these enzymes is protease inhibitor (PI). Plant protease inhibitor has tremendous potential as an antifungal agent which can be developed as biofungicide. In this work, protease inhibitors from B-serum (lutoid) of rubber tree latex were isolated and purified using Ion Exchange Chromatography (IEC) technique. Of the total 70 fractions of proteins extracted from the columns, only 26 fractions showed measurable levels of protein. The concentration of obtained putative protease inhibitors (three fractions of IEC) ranged from 0.007 to 0.022 mL/g B-serum. Inhibitory activity against four protease enzymes (subtilisin A, trypsin, α-chymotrypsin, and papain) showed the characteristics of Hevea putative protease inhibitors from B-serum as serine and/or cysteine protease inhibitors with more than 15% inhibitory activity of target protease. Based on SDS-PAGE visualization, the molecular weight of dominant protein considered as Hevea putative protease inhibitors was 21.5 kDa. In vitro bioassay test of antifungal activity for Hevea putative protease inhibitors showed reduced mycelium growth of Ganoderma boninense, Sclerotium sp., and Rigidosporus lignosus.[Keywords: protease inhibitor, Hevea brasiliensis, latex, B-serum, ion exchange chromatography]

scholarly journals The complex world of plant protease inhibitors: Insights into a Kunitz-type cysteine protease inhibitor of Arabidopsis thaliana

2017 ◽  
Vol 11 (1) ◽  
pp. e1368599 ◽  
Author(s):  
Sachin Rustgi ◽  
Edouard Boex-Fontvieille ◽  
Christiane Reinbothe ◽  
Diter von Wettstein ◽  
Steffen Reinbothe
Keyword(s):  
Arabidopsis Thaliana ◽  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Cysteine Protease ◽  
Cysteine Protease Inhibitor ◽  
Complex World ◽  
Plant Protease ◽  
Kunitz Type

scholarly journals A Review on Therapeutic Potential of Protease Inhibitors from Medicinal Plants

2019 ◽  
pp. 1-11
Author(s):  
R. N. N. Gamage ◽  
K. D. K. P. Kumari
Keyword(s):  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Blood Clotting ◽  
Therapeutic Potential ◽  
Biological Activities ◽  
Geographical Location ◽  
Therapeutic Agents ◽  
Human Diseases ◽  
Plant Protease ◽  
Protease Specificity

Most of the currently available therapeutic agents, particularly for cardiovascular disorders and cancers are very expensive and induce some serious side effects. Some of these drugs have also become less effective due to the emergence of antibiotic resistance. There is a necessity and great demand for the development of novel efficacious plant-based agents that are of pharmacologically effective. In this connection, this review focuses on therapeutic potential of plant protease inhibitors. Protease inhibitors are of a particular concern at present due to their potent ability to inhibit protease enzymes that are involved in pathogenesis of various human diseases. In addition to their function as protein-degrading enzymes, protease inhibitors are now well-known for their capability to involve in many biological activities as signaling molecules. Plant protease inhibitors are also engaged in several physiological and pathological processes, such as blood clotting, inflammation, immune regulation, apoptosis and carcinogenesis. Therefore, isolation of plant protease inhibitors and evaluation of their therapeutic capacity against chronic human diseases have become a major research interest. Nevertheless, protease inhibitor content and protease specificity vary significantly even in the same plant species depending on the geographical location and environmental factors. Consequently, it is important to identify potent therapeutic potential of each plant protease inhibitor on human health individually.

scholarly journals In vitro antioxidant extracts evaluation from the residue of the Hevea brasiliensis seed

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Giovanna Oleinik ◽  
Priscila Paola Dario ◽  
Katiane de Morais Gasperin ◽  
Dalila Moter Benvegnú ◽  
Fernanda Oliveira Lima ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Phenolic Compounds ◽  
Hevea Brasiliensis ◽  
Food Industry ◽  
Rubber Tree ◽  
Added Value ◽  
Future Research ◽  
Synthetic Antioxidants ◽  
Production Chain

AbstractThe antioxidants used in the food industry are essential to inhibit the formation of free radicals, preserving the existing properties in the different matrices. However, the insecurity of the synthetic antioxidants regarding human health propels search for natural substrates with potential antioxidant activity as an alternative to synthetic compounds. In this way, the work had as objective obtaining extracts from the seed pomace of the Hevea brasiliensis (rubber tree), relating the contents of flavonoids and total phenols in the application as an antioxidant. The methodology consisted of the extraction using four solvents, varying extractive methods, time, and seed concentrations. The antioxidant activity in vitro was evaluated by capturing the DPPH (2,2-diphenyl-1-picryl-hydrazil) radical. The optimized results demonstrate that the aqueous extracts produced in the Soxhlet in the concentrations of 85 g L−1 and retention time of 4 h reached 37.73 ± 1.69% in the antioxidant tests of the free radical DPPH capture, 1405.15 mg EAC 100 g−1 in the quantification of phenolic compounds and 223.34 mg 100 g−1 of total flavonoids. Thus, this work may contribute to the realization of studies and future research for characterization and identification concerning which phenolic compounds and flavonoids attribute the antioxidant characteristic to the extracts produced, enabling the discovery of products with high added value in the production chain. In addition, because the water used as a solvent showed greater antioxidant potential between the extracts, the non-toxic and environmentally friendly character is highlighted, allowing a wide variety of applications in the food industry.

scholarly journals A Phytophthora palmivora Extracellular Cystatin-Like Protease Inhibitor Targets Papain to Contribute to Virulence on Papaya

2018 ◽  
Vol 31 (3) ◽  
pp. 363-373 ◽  
Author(s):  
Rebecca Gumtow ◽  
Dongliang Wu ◽  
Janice Uchida ◽  
Miaoying Tian
Keyword(s):  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Cysteine Protease ◽  
Gene Editing ◽  
Sequence Data ◽  
Cysteine Protease Inhibitor ◽  
Phytophthora Palmivora ◽  
Cysteine Protease Inhibitors ◽  
Successful Infection

Papaya fruits, stems, and leaves are rich in papain, a cysteine protease that has been shown to mediate plant defense against pathogens and insects. Yet the oomycete Phytophthora palmivora is a destructive pathogen that infects all parts of papaya plants, suggesting that it has evolved cysteine protease inhibitors to inhibit papain to enable successful infection. Out of five putative extracellular cystatin-like cysteine protease inhibitors (PpalEPICs) from P. palmivora transcriptomic sequence data, PpalEPIC8 appeared to be unique to P. palmivora and was highly induced during infection of papaya. Purified recombinant PpalEPIC8 strongly inhibited papain enzyme activity, suggesting that it is a functional cysteine protease inhibitor. Homozygous PpalEPIC8 mutants were generated using CRISPR/Cas9-mediated gene editing via Agrobacterium-mediated transformation (AMT). Increased papain sensitivity of in-vitro growth and reduced pathogenicity during infection of papaya fruits were observed for the mutants compared with the wild-type strain, suggesting that PpalEPIC8, indeed, plays a role in P. palmivora virulence by inhibiting papain. This study provided genetic evidence demonstrating that plant-pathogenic oomycetes secrete cystatins as important weapons to invade plants. It also established an effective gene-editing system for P. palmivora by the combined use of CRISPR/Cas9 and AMT, which is expected to be applicable to other oomycetes.

In vitro mycorrhization of the rubber tree Hevea brasiliensis Müll Arg

2013 ◽  
Vol 49 (2) ◽  
pp. 207-215 ◽  
Author(s):  
Tiffany Sosa-Rodriguez ◽  
Hervé Dupré de Boulois ◽  
Françoise Granet ◽  
Sylvie Gaurel ◽  
Luz-Marina Melgarejo ◽  
...  
Keyword(s):  
Hevea Brasiliensis ◽  
Rubber Tree

In vitro photoautotrophic acclimatization, direct transplantation and ex vitro adaptation of rubber tree (Hevea brasiliensis)

2017 ◽  
Vol 133 (2) ◽  
pp. 215-223 ◽  
Author(s):  
Rujira Tisarum ◽  
Thapanee Samphumphung ◽  
Cattarin Theerawitaya ◽  
Wittaya Prommee ◽  
Suriyan Cha-um
Keyword(s):  
Hevea Brasiliensis ◽  
Rubber Tree ◽  
Ex Vitro

Purification and Characterization of a Protease Inhibitor with Anticancer Potential from Bacillus endophyticus JUPR15

2019 ◽  
Vol 15 (1) ◽  
pp. 74-82 ◽  
Author(s):  
Prerana Venkatachalam ◽  
Varalakshmi Kilingar Nadumane
Keyword(s):  
Protease Inhibitors ◽  
Inhibitory Activity ◽  
Main Function ◽  
Anticancer Compounds ◽  
Sds Page ◽  
Bacillus Endophyticus ◽  
Cold Acetone ◽  
Characterization Studies

Introduction:Introduction: Protease Inhibitors (PIs) constitute a group of proteins widely distributed among all organisms and their main function includes their ability to inhibit the proteolytic activity. PIs represent an important role in the regulation of various cellular physiological and biological processes, including cell cycle, cell death, differentiation and immune response.Material and Methods:Hence, in our search for novel anticancer compounds, we isolated microorganisms from various environmental sources and screened them for the production of protease inhibitors. Promising isolates were further checked for their protease inhibitory activity by their ability to inhibit the activity of trypsin and chymotrypsin, which were measured spectrophotometrically.Results:The isolate identified as Bacillus endophyticus JUPR15 was found to be promising with higher inhibitory activity than the other isolates. The inhibitor was purified by cold acetone precipitation and column chromatography and further subjected to characterization studies by performing 12 % SDS-PAGE to determine the molecular weight and gelatin-PAGE assay to confirm its inhibitory activity.Conclusion:The isolate exhibited promising anticancer activity on in-vitro Hela and HepG2 cancer cell lines, showing its application potentials.

Cloning and Characterization of Protease Inhibitor from Latex of Rubber Tree (Hevea brasiliensis)

2007 ◽  
Vol 08 (2) ◽  
pp. 12-17
Author(s):  
Aining Musor ◽  
Dr. Wilaiwan Chotigeat ◽  
Dr. Amornrat Phongdara
Keyword(s):  
Protease Inhibitor ◽  
Hevea Brasiliensis ◽  
Rubber Tree
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