scholarly journals BIOACTIVITY AND CLONING OF A NEW ANTIBACTERIAL LECTIN PROTEIN IN SPONGE Gelliodes sp. FROM BARANGLOMPO ISLAND IN SOUTH SULAWESI INDONESIA TERRESTRIAL

2010 ◽  
Vol 10 (2) ◽  
pp. 232-239
Author(s):  
Hanapi Usman ◽  
Prastawa Budi ◽  
Ahyar Ahmad

A research on antibacterial bioactivity of protein fraction isolated from several species of sponges of Barang Lompo Island has been conducted. Pre-purification of protein using fractional method, showed maximum bioactivity with the inhibition zone of 26 mm to Salmonella typhy from sponge Gelliodes sp. with the saturation level of ammonium sulfate of 40-60%. Further purification of this fraction using column chromatography followed by protein sequencing, indicated that pure protein as lectin, and behaves as a single-band on SDS-PAGE with molecular weight of 21 kDa. Based on amino acids partial sequence, we cloned and sequenced cDNA encoding lectin protein. It consists of 552 nucleotides encoding 183 amino acid residues including a putative initiation Met. To obtain it in large amounts, the coding sequence of lectin was cloned into pGEX-2TK vector and expression as a lectin fusion protein in Escherichia coli. Recombinant lectin exhibited a similar antibacterial activity to the native lectin. The recombinant lectin had stronger antibacterial activity toward S. typhy and S. aureus (G+) with the diameters of inhibition zone were 16 mm and 17 mm, respectively. This research might provide significant results for the following research on the antibacterial action in molecular level of lectin protein from marine sponges. Keywords: sponge, Gelliodes sp., lectin, Recombinant protein, antibacterial activity

2021 ◽  
Vol 948 (1) ◽  
pp. 012069
Author(s):  
R Trifani ◽  
Noverita ◽  
T A Hadi ◽  
E Sinaga

Abstract Endosymbiont fungi from marine sponges are a rich source of medicinally active compounds. Indonesia has a huge number of marine sponges. This research was conducted to determine the antibacterial potential of endosymbiont fungi isolated from marine sponges collected from Kotok Kecil Island, Seribu Islands. The screening was conducted with two methods, the modified GIBEX and disc diffusion. Species of marine sponges were isolated, namely Petrosia sp., Stylissa carteri, Cinachyrella australiensis, Callyspongia sp., Petrosia nigrians, and Stylissa massa, and obtained 9 isolates of endosymbiont fungi. The GIBEX test against Escherichia coli and Streptococcus mutans showed that ethyl acetate extract had strongest antibacterial activity. Disc diffusion test of ethyl acetate extract, five isolates had antibacterial activity against S. mutans and four against E. coli. The isolate that showed the strongest antibacterial activity was from Petrosia sp. Which has an inhibition zone of 8.4 mm against E. coli and 7.45 mm against S. mutans. The main active compounds from the isolate of Petrosia sp. are butylhydroxytoluene and phthalic acid di-(2-propylpentyl) esters. Based on this study, we concluded that the endosymbiont fungi of marine sponges are potential to be developed for further development as source of antibacterial agents.


2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Noor Hasniza Zin ◽  
Nurul Fatihah Ahmad ◽  
Noraslinda Muhamad Bunnori ◽  
Widya Abdul Wahab ◽  
Normah Haron

Introduction: Andrographis paniculata, Tinospora crispa and Centella asiatica are known to have various pharmacological functions. This research was carried out to investigate the antibacterial activities of protein extracts from A. paniculata, T. crispa and C. asiatica. Methods: Total soluble proteins from these herbs were extracted using a modified TCA/acetone method. The protein extracts were then quantified using the Bradford assay and separated using SDS-PAGE. The antibacterial activities were determined by disc diffusion method. Results: T. crispa had a significantly higher amount of proteins (83.86 ± 0.4 µg/µl) compared to A. paniculata (81.57 ± 0.4 µg/µl) and C. asiatica (78.93 ± 0.5 µg/µl). The proteins separated by SDS-PAGE were ranged from 30kDa to 260kDa, 25kDa to 110kDa and 25kDa to 160kDa for A. paniculata, T. crispa and C. Asiatic, respectively. The high abundance proteins were observed in A. paniculata and T. crispa but not in C. asitica. Protein extracts from C. asiatica have demonstrated antibacterial activity against all tested bacteria with the diameter of inhibition zone of 11.0 ± 0.5 mm, 12.3 ± 0.6 mm, 10.7 ± 0.7 mm and 20.0 ± 0.8 mm against B. cereus, S. aureus, K. pneumonia and S. typhimurium respectively. Meanwhile, protein extracts of A. paniculata showed a positive antibacterial activity only against B.cereus (13.7 ± 0.4 mm), S. aureus (7.0 ± 0.8 mm) and S. typhimurium (11.5 ± 0.3 mm). Protein extracts from T. crispa only showed a positive antibacterial activity against B. cereus (9.7 ± 0.5 mm). Conclusions: There is a constant need in the discovery of new antibiotics for the treatment of infectious diseases.


1997 ◽  
Vol 325 (3) ◽  
pp. 761-769 ◽  
Author(s):  
Isabelle GARCIA ◽  
Matthew RODGERS ◽  
Catherine LENNE ◽  
Anne ROLLAND ◽  
Alain SAILLAND ◽  
...  

p-Hydroxyphenylpyruvate dioxygenase catalyses the transformation of p-hydroxyphenylpyruvate into homogentisate. In plants this enzyme has a crucial role because homogentisate is the aromatic precursor of all prenylquinones. Furthermore this enzyme was recently identified as the molecular target for new families of potent herbicides. In this study we examine precisely the localization of p-hydroxyphenylpyruvate dioxygenase activity within carrot cells. Our results provide evidence that, in cultured carrot cells, p-hydroxyphenylpyruvate dioxygenase is associated with the cytosol. Purification and SDS/PAGE analysis of this enzyme revealed that its activity is associated with a polypeptide of 45–46 kDa. This protein specifically cross-reacts with an antiserum raised against the p-hydroxyphenylpyruvate dioxygenase of Pseudomonas fluorescens. Gel-filtration chromatography indicates that the enzyme behaves as a homodimer. We also report the isolation and nucleotide sequence of a cDNA encoding a carrot p-hydroxyphenylpyruvate dioxygenase. The nucleotide sequence (1684 bp) encodes a protein of 442 amino acid residues with a molecular mass of 48094 Da and shows specific C-terminal regions of similarity with other p-hydroxyphenylpyruvate dioxygenases. This cDNA encodes a functional p-hydroxyphenylpyruvate dioxygenase, as evidenced by expression studies with transformed Escherichia coli cells. Comparison of the N-terminal sequence of the 45–46 kDa polypeptide purified from carrot cells with the deduced peptide sequence of the cDNA confirms that this polypeptide supports p-hydroxyphenylpyruvate dioxygenase activity. Immunodetection studies of the native enzyme in carrot cellular extracts reveal that N-terminal proteolysis occurs during the process of purification. This proteolysis explains the difference in molecular masses between the purified protein and the deduced polypeptide.


Author(s):  
Eric Kwabena Droepenu ◽  
Ebenezer Aquisman Asare ◽  
Boon Siong Wee ◽  
Rafeah Binti Wahi ◽  
Frederick Ayertey ◽  
...  

Abstract Background Various parts of Anacardium occidentale plant possess curative qualities like antidiabetic, anti-inflammatory, antibacterial, antifungal, and antioxidant. Aqueous extract of this plant leaf was used in biosynthesizing zinc oxide (ZnO) nanoaggregates using two precursors of zinc salt (zinc acetate dihydrate [Zn(CH3COO)2∙2H2O] and zinc chloride [ZnCl2]). The synthesized ZnO samples were used in a comparative study to investigate the antibacterial activity against selected Gram-positive and Gram-negative microbes [Staphylococcus aureus, Exiguobacterium aquaticum (Gram +ve) and Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii (Gram −ve)]. The synthesized ZnO nanoaggregates from the two precursors were characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive x-ray spectroscopy (EDX) techniques. Results Micrographs of SEM and TEM confirmed nanoparticles agglomerated into aggregates. While spherical nanoaggregates were identified in samples prepared from Zn(CH3COO)2∙2H2O, flake-like structures were identified in samples synthesized from ZnCl2. Particle size determined by TEM was 107.03 ± 1.54 nm and 206.58 ± 1.86 nm for zinc acetate dihydrate and zinc chloride precursors respectively. ZnO nanoaggregate synthesized using zinc acetate as precursor gave higher antibacterial activity than its counterpart, zinc chloride with K. pneumonia recording the highest inhibition zone of 2.08 ± 0.03 mm (67.53%) whereas S. aureus recorded the least inhibition zone of 1.06 ± 0.14 mm (34.75%) for ZnO nanoaggregate from zinc chloride precursor. Also, antibacterial activity increases with increasing concentration of the extract in general. However, A. baumannii, E. aquaticum, and K. pneumoniae did not follow the continuity trend with regards to the 250 ppm and 500 ppm concentrations. Conclusion Biosynthesis of ZnO nanoaggregates using aqueous extract of A. occidentale leaf from zinc acetate dihydrate and zinc chloride as precursors was successful with the formation of nanospheres and nanoflakes. The study suggested that A. occidentale sp. could be an alternative source for the production of ZnO nanoparticles and are efficient antibacterial compounds against both Gram +ve and Gram −ve microbes with its promising effect against infectious bacteria.


2021 ◽  
Vol 17 ◽  
Author(s):  
Bambang Susilo ◽  
Abd. Rohim ◽  
Midia Lestari W. H.

Background: S. cristaefolium is the brown seaweed extracted using the serial technique with different solvents. Methods: S. cristaefolium powder (50 mesh) was extracted with hexane, ethyl acetate, and methanol respectively. The S. cristaefolium powder residue had been dried before being re-extracted with the next different solvents. Three serial extracts were obtained and named as the 1-stage extract, 2-stage extract, and 3-stage extract. Besides, a single-step extract (extraction using only methanol) was also produced to compare with three serial extracts in antibacterial activity tests (against E. coli and S. aureus). The three serial extracts were detected their antibacterial compounds using GC-MS, LC-HRMS, and FT-IR. Results: The 3-stage extract had the highest extraction yield. On S. aureus, the inhibition zone in all extracts was not significantly different. On E.coli, the highest inhibition zone (5.42±0.14 mm) was the 3-stage extract, indeed it is higher than both antibiotic and a single-step extract. Phenol, 9-Tricosene(Z)-, palmitic acid, and oleamide were contained in all extracts. Other antibacterial compound types, both the 1-stage and 2-stage extracts contained 8 types whilst the 3-stage extract contained the most types (12 types). Particularly, hexyl cinnamic aldehyde and betaine were detected only in the 3-stage extract with the dominant area. The carboxylic acid groups were detected in all extracts to confirm the fatty acid structure. Several cinnamic aldehyde groups were detected only in the 3-stage extract. Conclusions: Thus, the extraction technique serially could produce the 3-stage extract which has the strongest antibacterial activity and the richest antibacterial compounds.


Author(s):  
Sneka S ◽  
Preetha Santhakumar

Nano particles have an enormous impact on society. Selenium nanoparticles are used in various oxidative stresses. Capparis decidua is a plant which belongs to a family Capparidaceae. Capparis decidua is found in desert and semi desert areas and is used in Unani medicine and traditional system of medicine. The aim of the present study was to evaluate the antibacterial activity of selenium nanoparticles synthesized using Capparis decidua. Antibacterial activity was studied by inhibition zone against E.coli and Lactobacillus using Agar well diffusion method which was characterized by a clear zone. Selenium nanoparticles extracted from Capparis decidua fruit showed good antibacterial activity against lactobacillus species and E.coli.


2020 ◽  
Vol 6 (2) ◽  
pp. 246
Author(s):  
Yayuk Bulam Sarifati ◽  
Sjarif Ismail ◽  
Khemasili Kosala

Mekai leaves (Pycnarrhena cauliflora Diels.) (P. cauliflora). Are known to contain flavonoid compounds, tannins and phenolics that act as antibacterial agents and are used in the treatment of eye pain. Staphylococcus aureus (S. aureus) is one of the examples of bacterial diseases of eye pain and also a major cause of many infections in communities and health facilities with cases of resistance to various antimicrobial agents. The purpose of this study was to prove the antibacterial activity of mekai leaves ethanol extract against S. aureus bacteria. This research is an experimental research. The stages of this research began by extracting mekai leaves using maceration method with 96% ethanol solvent. Antibacterial activity was tested by the disc method (Kirby-Bauer) using ethanol extract concentrations of mekai leaves (EPC) 20%, 30%, 40%, 50%, 60%, 70%, and 80%, positive control using 25 μg amoxicillin and negative control using DMSO 10%. The measurement results of inhibition zones of mekai leaf ethanol extract 20%, 30%, 40%, 50%, 60%, 70% and 80% respectively were 8.32 mm, 8.32 mm, 8.32 mm, 8.67 mm, 9.00 mm, 8.67 mm, and 8.33 mm. While the positive control measurement of 25 μg amoxicillin against S. aureus is 28.67 mm and the measurement of 10% negative DMSO control does not produce inhibitory zones, so it can be concluded that the ethanol extract of mekai leaves has antibacterial activity, but the area of ​​inhibition zone produced is smaller than amoxicillin 25 µg. The results of statistical tests using Mann Whitney between negative controls with all EPC concentrations obtained significant differences with p values ​​<0.05, it can be concluded that there is antibacterial activity produced at all EPC concentrations.


2020 ◽  
Vol 11 (2) ◽  
pp. 35-41
Author(s):  
Callixte Yadufashije ◽  
Adolyne Niyonkuru ◽  
Emanuel Munyeshyaka ◽  
Sibomana Madjidi ◽  
Joseph Mucumbitsi

Background: Ginger (Zingiber officinale) has been used for long time due to its potential antimicrobial activity against diversity of microbial pathogens. Aims and Objectives: The study was carried out to investigate the bacteria pathogens found in digestive tract infections and assess antimicrobial activities of ginger extract to identified bacteria. Materials and Methods: Bacteriological studies were carried out on stool samples from 30 patients attending Muhoza health center. Different types of bacteria were isolated from stool samples of digestive tract infection patients by using various methods such culture, biochemical test and antimicrobial activity of ginger extracts was analyzed at INES-Ruhengeri in clinical microbiology laboratory. Results: Study findings showed isolated bacteria and antibacterial activity of Ginger. Isolated bacteria and their percentages including Escherichia coli (46.6%) which is the predominant isolated bacteria, Salmonella species (33.33%), Enterobacter spp (10.0%), Shigella spp (6.6%) and Citrobacter (3.33%) which is the least isolated bacteria. Antibacterial activity of ginger was seen on isolated bacteria, as ethanol and methanol were used for ginger oil extraction, the antibacterial activity of ginger extracts using ethanol was seen on isolated bacteria such us Citrobacter spp with 14 mm of inhibition zone, Shigella spp with 12 mm, Salmonella with 11.1 mm, E. coli with 9.5 mm and Enterobacter spp which was seen to be resistant to ginger extract using ethanol with 0.66mm of inhibition zone. For methanol extracts antibacterial activity was seen as follows: Citrobacter spp at 12 mm, Shigella spp at 11 mm, E. coli at 8 mm, salmonella spp at 6.1 mm, and Enterobacter spp with 5 mm. Enterobacter spp was seen to be the most resistant bacteria in both extracts. Conclusion: Ginger has shown to have an antibacterial activity on bacteria isolated from digestive tract infected patients. It can be used as a medicine to treat these infections. Number of researches should be done to be sure on this reality of antibacterial activity of ginger.


2019 ◽  
Vol 10 (2) ◽  
pp. 94-98
Author(s):  
Adlis Santoni ◽  
Mai Efdi ◽  
Akmel Suhada

Hornstedtia scyphifera var. fusiformis Holttum is endemic plant of Zingiberaceae family that distributed in Sumatera. Hornstedtia scyphifera var. fusiformis Holttum contained some secondary metabolite such as phenolic, saponin, triterpenoid and alkaloid. The biological activity of methanol, ethyl acetate, and n-hexane extracts from Hornstedtia scyphifera var. fusiformis Holttum suspended roots was tested for antibacterial and antioxydant activity. Antioxidant activity was analized by DPPH method. The antibacterial activity  was tested by used disk difussion method againts Staphylococcus aureus bacteria (gram-positive bacteria) and Escherichia coli (gram-negative bacteria). Almost all of Hornstedtia scyphifera var. fusiformis Holttum suspended roots extract were given antibacterial activity, nonetheless the biggest inhibition zone of Escherichia coli that was inhibited by n-hexane and ethyl acetate extracts which have inhibition zone 10 mm at  concentration 500 mg/L and also againts Staphylococcus aureus, the biggest inhibition by ethyl acetate and n-hexane extracts which have inhibition zone 10.30 mm at concentration 500 mg/L. Among all extracts tested, methanol extract of the possessed moderate free radical scavenging activity with IC50 = 51.89 mg/L.


2016 ◽  
Vol 12 (1) ◽  
pp. 1
Author(s):  
Pramudita Putri Kusuma ◽  
Ganjar Fadillah ◽  
Husna Syaima ◽  
Teguh Endah Saraswati

<p>The addition of garlic powder to gelatin from chicken claw waste was potentially developed as a natural preservative in food, especially for meat. Preparation of gelatin/garlic biocomposite was performed in three stages: synthesis of gelatin from chicken claw, garlic powder preparation as allicin source and preparation of biocomposite gelatin/garlic. The preparation of dry biocomposites was done by weighing the gelatin and garlic powder in weight ratio of 1 : 1 and 1 : 2 (w/w) in the total mass of 0.75 grams. For wet biocomposite preparation, the mixture of the powder was solved in 5 mL of lactic acid 2 %. Functional groups of gelatin, garlic and biocomposite were analyzed by <em>f</em><em>ourier transform infrared spectroscopy</em> (FTIR). The antibacterial activity of biocomposite against <em>Staphylococcus aureus</em> were tested using disc diffusion method. This test was performed on garlic powder, solvent and gelatin/garlic biocomposites powder in the ratio of 1 : 1 and 1 : 2 in 2 % lactic acid solvent. The biocomposite with a weight ratio of gelatin : garlic of  1 : 1 had the optimum diameter of inhibition zone. The effectiveness of biocomposite gelatin/garlic as natural preservative applied in meat was also physically studied by organoleptic analysis. Organoleptic analysis through the hedonic test was conducted on the parameters of color, smell, and texture of gelatin/garlic biocomposites-coated meat. The results showed that the addition of garlic can increase the effectiveness of gelatin as a natural preservative of meat for four days stored in closed packaging at room temperature.</p>


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