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Fulltext ACE inhibitory activity of pangasius catfish (Pangasius sutchi) skin and bone gelatin hydrolysate

Mahmoodani F, Ghassem M, Babji AS, Yusop SM, Khosrokhavar R

J Food Sci Technol, 2014 Sep;51(9):1847-56.
PMID: 25190839 DOI: 10.1007/s13197-012-0742-8

Skin and bone gelatins of pangasius catfish (Pangasius sutchi) were hydrolyzed with alcalase to isolate Angiotensin Converting Enzyme (ACE) inhibitory peptides. Samples with the highest degree of hydrolysis (DH) were separated into different fractions with molecular weight cut-off (MWCO) sizes of 10, 3 and 1 kDa, respectively and assayed for ACE inhibitory activity. Skin and bone gelatins had highest DH of 64.87 and 68.48 % after 2 and 1 h incubation, respectively. Results from this study indicated that by decreasing the molecular weight of fractions, ACE inhibitory activity was increased. Therefore, F3 permeates (MWCO

Matched MeSH terms: Peptide Hydrolases
The identification of active compounds in Ganoderma lucidum var. antler extract inhibiting dengue virus serine protease and its computational studies

Lim WZ, Cheng PG, Abdulrahman AY, Teoh TC

J Biomol Struct Dyn, 2020 Sep;38(14):4273-4288.
PMID: 31595837 DOI: 10.1080/07391102.2019.1678523

The number of global dengue incidences is alarmingly high in recent years. The global distribution of four dengue serotypes has also added economic burden in the dengue-endemic countries. To discover the potent dengue virus inhibitors in the antler form of Ganoderma lucidum (Lingzhi or Reishi), the water extraction of normal G. lucidum and its antler form were conducted and the chemical compounds were identified by LC-MS. Six distinct chemical compounds identified in high abundance were hesperetin, thymidine, lucidenic acid, 11-aminoundecanoic acid, 5-carboxyvanillic acid and ganocin B. The water extracts of G. lucidum in its antler form inhibited the DENV2 NS2B-NS3 protease activity at 84.6 ± 0.7%, higher than the normal G. lucidum. Then, molecular docking was performed on the homology model built from an in-house sequence. Docking simulation results showed that hesperetin and ganocin B were the best leads to bind at the catalytic triad of DENV2 NS2B-NS3pro via hydrogen bonding, van der Waals and pi-pi interactions. Extensive overlapping of HOMO-LUMO orbitals at the ringed regions of hesperetin helped to facilitate the entry of ligand to the catalytic triad cleft. LC-MS, molecular docking and density functional theory analyses confirmed that hesperetin was the strongest inhibitor against NS2B-NS3 protease. Communicated by Ramaswamy H. Sarma.

Matched MeSH terms: Peptide Hydrolases
Fulltext A Review on the Biotechnological Applications of the Operational Group Bacillus amyloliquefaciens

Ngalimat MS, Yahaya RSR, Baharudin MMA, Yaminudin SM, Karim M, Ahmad SA, et al.

Microorganisms, 2021 Mar 17;9(3).
PMID: 33802666 DOI: 10.3390/microorganisms9030614

Bacteria under the operational group Bacillus amyloliquefaciens (OGBa) are all Gram-positive, endospore-forming, and rod-shaped. Taxonomically, the OGBa belongs to the Bacillus subtilis species complex, family Bacillaceae, class Bacilli, and phylum Firmicutes. To date, the OGBa comprises four bacterial species: Bacillus amyloliquefaciens, Bacillus siamensis, Bacillus velezensis and Bacillus nakamurai. They are widely distributed in various niches including soil, plants, food, and water. A resurgence in genome mining has caused an increased focus on the biotechnological applications of bacterial species belonging to the OGBa. The members of OGBa are known as plant growth-promoting bacteria (PGPB) due to their abilities to fix nitrogen, solubilize phosphate, and produce siderophore and phytohormones, as well as antimicrobial compounds. Moreover, they are also reported to produce various enzymes including α-amylase, protease, lipase, cellulase, xylanase, pectinase, aminotransferase, barnase, peroxidase, and laccase. Antimicrobial compounds that able to inhibit the growth of pathogens including non-ribosomal peptides and polyketides are also produced by these bacteria. Within the OGBa, various B. velezensis strains are promising for use as probiotics for animals and fishes. Genome mining has revealed the potential applications of members of OGBa for removing organophosphorus (OPs) pesticides. Thus, this review focused on the applicability of members of OGBa as plant growth promoters, biocontrol agents, probiotics, bioremediation agents, as well as producers of commercial enzymes and antibiotics. Here, the bioformulations and commercial products available based on these bacteria are also highlighted. This review will better facilitate understandings of members of OGBa and their biotechnological applications.

Matched MeSH terms: Peptide Hydrolases
Fulltext Identification of Potential Inhibitors of 3CL Protease of SARS-CoV-2 From ZINC Database by Molecular Docking-Based Virtual Screening

Abdusalam AAA, Murugaiyah V

Front Mol Biosci, 2020;7:603037.
PMID: 33392261 DOI: 10.3389/fmolb.2020.603037

The rapid outbreak of Coronavirus Disease 2019 (COVID-19) that was first identified in Wuhan, China is caused by a novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The 3CL protease (3CLpro) is the main protease of the SARS-CoV-2, which is responsible for the viral replication and therefore considered as an attractive drug target since to date there is no specific and effective vaccine available against this virus. In this paper, we reported molecular docking-based virtual screening (VS) of 2000 compounds obtained from the ZINC database and 10 FDA-approved (antiviral and anti-malaria) on 3CLpro using AutoDock Vina to find potential inhibitors. The screening results showed that the top four compounds, namely ZINC32960814, ZINC12006217, ZINC03231196, and ZINC33173588 exhibited high affinity at the 3CLpro binding pocket. Their free energy of binding (FEB) were -12.3, -11.9, -11.7, and -11.2 kcal/mol while AutoDock Vina scores were -12.61, -12.32, -12.01, and -11.92 kcal/mol, respectively. These results were better than the co-crystallized ligand N3, whereby its FEB was -7.5 kcal/mol and FDA-approved drugs. Different but stable interactions were obtained between the four identified compounds with the catalytic dyad residues of the 3CLpro. In conclusion, novel 3CLpro inhibitors from the ZINC database were successfully identified using VS and molecular docking approach, fulfilling the Lipinski rule of five, and having low FEB and functional molecular interactions with the target protein. The findings suggests that the identified compounds may serve as potential leads that act as COVID-19 3CLpro inhibitors, worthy for further evaluation and development.

Matched MeSH terms: Peptide Hydrolases
Plant growth promoting and antifungal asset of indigenous rhizobacteria secluded from saffron (Crocus sativus L.) rhizosphere

Rasool A, Imran Mir M, Zulfajri M, Hanafiah MM, Azeem Unnisa S, Mahboob M

Microb Pathog, 2021 Jan;150:104734.
PMID: 33429050 DOI: 10.1016/j.micpath.2021.104734

Saffron (Crocus sativus L.) is an important plant in medicine. The Kashmir Valley (J&K, India) is one of the world's largest and finest saffron producing regions. However, over the past decade, there has been a strong declining trend in saffron production in this area. Plant Growth Promoting Rhizobacteria (PGPR) are free living soil bacteria that have ability to colonize the surfaces of the roots and ability to boost plant growth and development either directly or indirectly. Using the efficient PGPR as a bio-inoculant is another sustainable agricultural practice to improve soil health, grain yield quality, and biodiversity conservation. In the present study, a total of 13 bacterial strains were isolated from rhizospheric soil of saffron during the flowering stage of the tubers and were evaluated for various plant growth promoting characteristics under in vitro conditions such as the solubilization of phosphate, production of indole acetic acid, siderophore, hydrocyanic acid, and ammonia production and antagonism by dual culture test against Sclerotium rolfsii and Fusarium oxysporum. All the isolates were further tested for the production of hydrolytic enzymes such as protease, lipase, amylase, cellulase, and chitinase. The maximum proportions of bacterial isolates were gram-negative bacilli. About 77% of the bacterial isolates showed IAA production, 46% exhibited phosphate solubilization, 46% siderophore, 61% HCN, 100% ammonia production, 69% isolates showed protease activity, 62% lipase, 46% amylase, 85% cellulase, and 39% showed chitinase activity. Three isolates viz., AIS-3, AIS-8 and AIS-10 were found to have the most plant growth properties and effectively control the growth of Sclerotium rolfsii and Fusarium oxysporum. The bacterial isolates were identified as Brevibacterium frigoritolerans (AIS-3), Alcaligenes faecalis subsp. Phenolicus (AIS-8) and Bacillus aryabhattai (AIS-10) respectively by 16S rRNA sequence analysis. Therefore, these isolated rhizobacterial strains could be a promising source of plant growth stimulants to increase cormlets growth and increase saffron production.

Matched MeSH terms: Peptide Hydrolases
Fulltext Prevalence, virulence and antifungal activity of C. albicans isolated from infected root canals

Abraham SB, Al Marzooq F, Himratul-Aznita WH, Ahmed HMA, Samaranayake LP

BMC Oral Health, 2020 12 01;20(1):347.
PMID: 33256696 DOI: 10.1186/s12903-020-01347-5

BACKGROUND: There is limited data on the prevalence of Candida species in infected root canal systems of human teeth. We attempted to investigate the prevalence, genotype, virulence and the antifungal susceptibility of Candida albicans isolated from infected root canals of patients with primary and post-treatment infections in a UAE population.
METHODS: Microbiological samples from 71 subjects with infected root canals were aseptically collected, and cultured on Sabouraud dextrose agar, and C. albicans was identified using multiplex polymerase chain reaction, and the isolates were further subtyped using ABC genotyping system. Their relative virulence was compared using further four archival samples of endodontic origin from another geographical region, and four more salivary isolates, as controls. The virulence attributes compared were biofilm formation, and production of phospholipase and haemolysin, and the susceptibility to nystatin, amphotericin B, ketoconazole, and fluoconazole was also tested.
RESULTS: 4 out of 71 samples (5.6%) yielded Candida species. On analysis of variance among the groups, the intracanal isolates, mainly Genotype A, possessed a high degree of phospholipase and haemolysin activity (p

Matched MeSH terms: Hydrolases
Fulltext Toxicity Evaluation of the Naphthalen-2-yl 3,5-Dinitrobenzoate: A Drug Candidate for Alzheimer Disease

Anwar F, Saleem U, Rehman AU, Ahmad B, Froeyen M, Mirza MU, et al.

Front Pharmacol, 2021;12:607026.
PMID: 34040515 DOI: 10.3389/fphar.2021.607026

The presented study was designed to probe the toxicity potential of newly identified compound naphthalen-2-yl 3,5-dinitrobenzoate (SF1). Acute, subacute toxicity and teratogenicity studies were performed as per Organization of economic cooperation and development (OECD) 425, 407, and 414 test guidelines, respectively. An oral dose of 2000 mg/kg to rats for acute toxicity. Furthermore, 5, 10, 20, and 40 mg/kg doses were administered once daily for 28 days in subacute toxicity study. Teratogenicity study was performed with 40 mg/kg due to its excellent anti-Alzheimer results at this dose. SF1 induced a significant rise in Alkaline Phosphatases (ALP), bilirubin, white blood cells (WBC), and lymphocyte levels with a decrease in platelet count. Furthermore, the reduction in urea, uric acid, and aspartate transaminase (AST) levels and an increase in total protein levels were measured in subacute toxicity. SF1 increased spermatogenesis at 5 and 10 mg/kg doses. Teratogenicity study depicted no resorptions, early abortions, cleft palate, spina bifida and any skeletal abnormalities in the fetuses. Oxidative stress markers (Superoxide dismutase (SOD), Catalase (CAT), and glutathione (GSH) were increased in all the experiments, whereas the effect on melanoaldehyde Malondialdehyde (MDA) levels was variable. Histopathology further corroborated these results with no change in the architectures of selected organs. Consequently, a 2000 mg/kg dose of SF1 tends to induce minor liver dysfunction along with immunomodulation, and it is well below its LD
50
. Moreover, it can be safely used in pregnancy owing to its no detectable teratogenicity.

Matched MeSH terms: Phosphoric Monoester Hydrolases
Comparative proteomes, immunoreactivities and neutralization of procoagulant activities of Calloselasma rhodostoma (Malayan pit viper) venoms from four regions in Southeast Asia

Tang ELH, Tan NH, Fung SY, Tan CH

Toxicon, 2019 Aug 22;169:91-102.
PMID: 31445943 DOI: 10.1016/j.toxicon.2019.08.004

The intraspecific geographical venom variations of Calloselasma rhodostoma from Malaysia (CR-M), Indonesia (CR-I), Thailand (CR-T) and Vietnam (CR-V) were investigated through 1D SDS-PAGE and nano-ESI-LCMS/MS. The venom antigenicity, procoagulant activities and neutralization using Thai C. rhodostoma Monovalent Antivenom (CRMAV) were also investigated. SDS-PAGE patterns of the venoms were relatively similar with minor variations. Proteomic analysis revealed that snake venom metalloproteinases (SVMPs, particularly P-I class), serine proteases (SVSPs) and snaclecs dominated the venom protein composition (68.96-81.80%), followed by L-amino acid oxidase (LAAO) and phospholipase A2 (PLA2) (7.37-11.08% and 5.18-13.81%, respectively), corroborating C. rhodostoma envenoming effects (hemorrhage, consumptive coagulopathy, thrombocytopenia and local tissue necrosis). Other proteins of lower abundances (2.82-9.13%) identified include cysteine-rich secretory proteins (CRISP), phospholipase B, phosphodiesterase, nerve growth factor, 5'-nucleotidase, aminopeptidase and hyaluronidase. All four venoms exhibited strong procoagulant effects which were neutralized by CRMAV to different extents. CRMAV immunoreactivity was high toward venoms of CR-M, CR-I and CR-T but relatively low for CR-V venom. Among the venom samples from different locales, CR-V venom proteome has the smallest SVMP composition while SVSP, PLA2 and phosphodiesterase were more abundant in the venom. These variations in C. rhodostoma venom protein composition could partly explain the differences seen in immunoreactivity. (198 words).

Matched MeSH terms: Phosphoric Diester Hydrolases
Fulltext Proteinases in Naegleria Fowleri (strain NF3), a pathogenic amoeba: a preliminary study

Mat Amin N

Trop Biomed, 2004 Dec;21(2):57-60.
PMID: 16493399

Naegleria fowleri is a free-living amoeba, known as a causative agent for a fatal disease of the central nervous system (CNS) in man such as Primary amoebic meningoencephalitis (PAM). Factors contributing to its pathogenicity and its distribution in the environment have been investigated by previous researchers. In case of its pathogenicity, several enzymes such as phospolipase A and sphingomyelinase, have been proposed to probably act as aggressors in promoting PAM but no study so far have been conducted to investigate the presence of proteinase enzyme in this amoeba although a 56kDa cystein proteinase enzyme has been identified in Entamoeba histolytica as an important contributing factor in the amoeba's virulence. In this preliminary study, a pathogenic amoeba, Naegleria fowleri (strain NF3) was examined for the presence of proteinases. Samples of enzymes in this amoeba were analysed by electrophoresis using SDS-PAGE-gelatin gels. The results showed that this amoeba possesses at least two high molecular weight proteinases on gelatin gels; their apparent molecular weights are approximately 128 kDa and approximately 170 kDa. Band of approximately 128 kDa enzyme is membrane-associated and its activity is higher at alkaline pH compared with lower pH; at lower pH, its activity is greatly stimulated by DTT. The approximately 170 kDa band enzyme appears to be inactivated at pH 8.0, at lower ph its activity is higher and DTT-dependance. The activity of this enzyme is partially inhibited by inhibitor E-64 but markedly inhibited to antipain suggesting it belongs to the cysteine proteinase group.

Matched MeSH terms: Peptide Hydrolases
Prediction of promiscuous epitopes in NSP2 of Chikungunya virus: An in-silico approach

Fazal F, Anwar T, Waheed Y, Parvaiz F

Trop Biomed, 2020 Sep 01;37(3):566-577.
PMID: 33612772 DOI: 10.47665/tb.37.3.566

This study is focused towards developing a global consensus sequence of nonstructural protein 2 (NSP2), a protease of Chikungunya Virus (CHIKV) and predict immunogenic promiscuous T-cell epitopes based on various bioinformatics tools. To date, no epitope data is available for the Chikungunya virus in the IEDB database. In this study, 100 available nucleotide sequences of NSP2-CHIKV belonging to different strains were downloaded from the National Centre for Biotechnology Information (NCBI) database. The nucleotide sequences were subjected to translated sequencing using the EXPASY tool followed by protein alignment using the CLC workbench and a global consensus sequence for the respective protein was developed. IEDB tool was used to predict HLA-I and HLA-II binding promiscuous epitopes from the consensus sequence of NSP2-CHIKV. Thirty-four B-cell based epitopes are predicted and the promiscuous epitope is VVDTTGSTKPDPGD at position 341-354. Twenty-six MHC-I short peptide epitopes are predicted to bind with HLA-A. The promiscuous epitopes predicted to bind with HLA-A*01:01 are VTAIVSSLHY, SLSESATMVY, FSKPLVYY, QPTDHVVGEY at positions 317-326, 84-93, 535-544 and 15-24 with percentile ranks 0.17, 0.39, 0.51 and 0.81, respectively. Twenty-four MHC-II short peptide epitopes are predicted for HLA-DRB. The promiscuous epitope predicted to bind with HLA-DRB*01:01 is VVGEYLVLSPQTVLRS from 20-35 with a lowest percentile rank of 0.01. These predicted epitopes can be effective targets towards development of vaccine against CHIKV. Epitopes predicted in this study displayed good binding affinity, antigenicity and promiscuity for the HLA classes. These predicted epitopes can prove to be translationally important towards the development of CHIKV.

Matched MeSH terms: Peptide Hydrolases
Fulltext Properties of proteolytic enzyme from ginger (Zingiber officinale Roscoe)

Nafi’, A., Foo, H.L., Jamilah, B., Ghazali. H.M.

International Food Research Journal, 2013;20(1):363-368.
MyJurnal

Proteases in ginger rhizome have the potentials in industrial applications. This study was conducted to extract and characterize the proteolytic enzyme from ginger (Zingiber officinale Roscoe). Ginger protease (GP) was extracted from ginger rhizome by homogenization with 100 mM potassium phosphate buffer pH 7.0 containing 10 mM cysteine and 5 mM EDTA which were found to be the most efficient extraction buffer and stabilizers. After centrifugation at 10,500 x g, protein in the crude extract was precipitated using 60% ammonium sulfate following which the precipitate was redissolved in 50 mM potassium phosphate buffer pH 7.0, dialyzed and then lyophilized. The extraction method yielded 0.94% (w/w of fresh weight) of GP with a specific activity of 27.6 ± 0.1 Unit/mg protein where 1 Unit is defined as the amount of protease causing an increase in absorbance by 1 unit per minute using azocasein as the substrate. Results show that the GP was completely inhibited by heavy metal cations i.e. Cu2+and Hg2+, and a thiol blocking agent or inhibitor, n-ethyl maleimide (NEM), indicating that GP is most probably a cysteine protease. The enzyme has an optimum temperature at 60⁰C and the optimum pH ranged between pH 6 to 8. Monovalent cations (K+ and Na+) have no significant effect on activity of GP, but divalent and trivalent cations showed moderate inhibitory effect. Detergents such as sodium dodecyl sulfate increased the activity of GP while Tween 80 and Tween 20 slightly reduced the activity.

Matched MeSH terms: Peptide Hydrolases
Fulltext Pentocin MQ1: A Novel, Broad-Spectrum, Pore-Forming Bacteriocin From Lactobacillus pentosus CS2 With Quorum Sensing Regulatory Mechanism and Biopreservative Potential

Wayah SB, Philip K

Front Microbiol, 2018;9:564.
PMID: 29636737 DOI: 10.3389/fmicb.2018.00564

Micrococcus luteus, Listeria monocytogenes, and Bacillus cereus are major food-borne pathogenic and spoilage bacteria. Emergence of antibiotic resistance and consumer demand for foods containing less of chemical preservatives led to a search for natural antimicrobials. A study aimed at characterizing, investigating the mechanism of action and regulation of biosynthesis and evaluating the biopreservative potential of pentocin from Lactobacillus pentosus CS2 was conducted. Pentocin MQ1 is a novel bacteriocin isolated from L. pentosus CS2 of coconut shake origin. The purification strategy involved adsorption-desorption of bacteriocin followed by RP-HPLC. It has a molecular weight of 2110.672 Da as determined by MALDI-TOF mass spectrometry and a molar extinction value of 298.82 M-1 cm-1. Pentocin MQ1 is not plasmid-borne and its biosynthesis is regulated by a quorum sensing mechanism. It has a broad spectrum of antibacterial activity, exhibited high chemical, thermal and pH stability but proved sensitive to proteolytic enzymes. It is potent against M. luteus, B. cereus, and L. monocytogenes at micromolar concentrations. It is quick-acting and exhibited a bactericidal mode of action against its targets. Target killing was mediated by pore formation. We report for the first time membrane permeabilization as a mechanism of action of the pentocin from the study against Gram-positive bacteria. Pentocin MQ1 is a cell wall-associated bacteriocin. Application of pentocin MQ1 improved the microbiological quality and extended the shelf life of fresh banana. This is the first report on the biopreservation of banana using bacteriocin. These findings place pentocin MQ1 as a potential biopreservative for further evaluation in food and medical applications.

Matched MeSH terms: Peptide Hydrolases
Fulltext Brain Lipopolysaccharide Preconditioning-Induced Gene Reprogramming Mediates a Tolerance State in Electroconvulsive Shock Model of Epilepsy

Amini E, Golpich M, Farjam AS, Kamalidehghan B, Mohamed Z, Ibrahim NM, et al.

Front Pharmacol, 2018;9:416.
PMID: 29765321 DOI: 10.3389/fphar.2018.00416

There is increasing evidence pointing toward the role of inflammatory processes in epileptic seizures, and reciprocally, prolonged seizures induce more inflammation in the brain. In this regard, effective strategies to control epilepsy resulting from neuroinflammation could be targeted. Based on the available data, preconditioning (PC) with low dose lipopolysaccharide (LPS) through the regulation of the TLR4 signaling pathway provides neuroprotection against subsequent challenge with injury in the brain. To test this, we examined the effects of a single and chronic brain LPS PC, which is expected to lead to reduction of inflammation against epileptic seizures induced by electroconvulsive shock (ECS). A total of 60 male Sprague Dawley rats were randomly assigned to five groups: control, vehicle (single and chronic), and LPS PC (single and chronic). We first recorded the data regarding the behavioral and histological changes. We further investigated the alterations of gene and protein expression of important mediators in relation to TLR4 and inflammatory signaling pathways. Interestingly, significant increased presence of NFκB inhibitors [Src homology 2-containing inositol phosphatase-1 (SHIP1) and Toll interacting protein (TOLLIP)] was observed in LPS-preconditioned animals. This result was also associated with over-expression of IRF3 activity and anti-inflammatory markers, along with down-regulation of pro-inflammatory mediators. Summarizing, the analysis revealed that PC with LPS prior to seizure induction may have a neuroprotective effect possibly by reprogramming the signaling response to injury.

Matched MeSH terms: Phosphoric Monoester Hydrolases
Penghasilan Sirap Glukosa Berfruktosa Tinggi dari Kanji Ubi Keledek Msp94 (Production of High Fructose Glucose Syrup from Sweet Potato MSP94 Starch)

Hasnah Haron, Osman Hassan, Mamot Said

Sains Malaysiana, 2005;34.

This study comprised of physicochemical characterizations of starch extracted from Msp94 sweet potato tuber and production of high fructose glucose syrup from the starch. Msp94 sweet potato starch consisted of 7.3% water, 0.2% protein, 0.4% fat, 1.3% total ash, 94.8% total carbohydrates, 83.0% starch and 20.6% apparent amylose. The starch granules were spherical, polygonal and irregular in shapes with the size of 13-14 mm. Enzymatic hydrolysis of Msp94 sweet potato starch for 24,48, 72 hours, using a mixture of amyglucosidase-pullulanase enzymes during saccharification process, produced starch hydrolysates with dextrose equivalent (DE) of 94.8, 99.1, 99.3 respectively. This is followed by reduction in viscosity of the starch hydrolysates. Conversion of the Msp94 starch to percent of glucose after hydrolysing for 24,48 and 72 hours were 97.1%, 109.5% and 103.2%, respectively. Msp94 starch hydrolysates was then purified using three types of ion exchange resins and isomerized to highfructose syrup using glucose isomerase enzyme (Sweetzyme T). Thefructose content in isomerized Msp94 syrup was (43.8-46.5%) was comparable to the fructose content (44%) in commercial High Fructose Corn Syrup (HFCS) 42.
[Kajian ini merangkumi pencirian fizikokimia kanji yang diekstrak daripada ubi keledek Msp94 dan penghasilan sirap glukosa berfruktosa tinggi daripada kanji ini. Kanji ubi keledek Msp94 mengandungi 7.3% air, 0.2% protein, 0.4% lemak, 1.3% abu total, 94.8% karbohidrat total, 83.0% kanji dan 20.6% amilosa ketara. Purata saiz granul kanji adalah 13-14 mm, berbentuk bulat, poligon dan bentuk yang tidak tetap. Hidrolisis berenzim menggunakan gabungan enzim glukoamilase-pululanase dalam proses sakarifikasi, yang dijalankan ke atas kanji ubi keledek Msp94 selama 24, 48, 72 jam menghasilkan hidrolisat kanji dengan setaraan dekstrosa (DE) masing-masing pada 94.8, 99.1, 99.3. 1ni diikuti dengan kelikatan hidrolisat kanji yang semakin menurun. Penukaran kanji Msp94 kepada peratus glukosa adalah sebanyak 97.1 %, 109.5% dan 103.2% setelah dihidrolisis selama 24,48 dan 72 jam. Hidrolisat kanji Msp94 ditulenkan menggunakan tiga jenis resin penukar ion dan diisomer kepada sirap berfruktosa tinggi menggunakan enzim glukosa isomerase (Sweetzyme T). Kandungan fruktosa (43.8-46.5%) dalam sirap Msp94 yang telah diisomer adalah setara dengan kandunganfruktosa (44%) dalam sirap komersial, High Fructose Corn Syrup (HFCS) 42].

Matched MeSH terms: Glycoside Hydrolases
Proteomes of the female salivary glands of Simulium nigrogilvum and Simulium nodosum, the main human-biting black flies in Thailand

Hempolchom C, Reamtong O, Sookrung N, Srisuka W, Sakolvaree Y, Chaicumpa W, et al.

Acta Trop, 2019 Jun;194:82-88.
PMID: 30922801 DOI: 10.1016/j.actatropica.2019.03.026

Although several studies have reported pharmacological and immunological activity, as well as the role of black flies in transmitting pathogens to vertebrate hosts through salivary glands (SG) during blood feeding, SG proteomes of the anthropophilic black flies in Thailand have never been reported. Therefore, this study determined the SG proteomes of female S. nigrogilvum and S. nodosum. Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional (2-DE) gels containing separated SG proteins of individual species were subjected to liquid chromatography-tandem mass spectrometry (LCMS/MS) and an orthologous protein search from eukaryotic organism, nematocera and simuliidae databases for total protein identification. SDS-PAGE and protein staining revealed at least 13 and 9 major protein bands in the SGs of female S. nigrogilvum and S. nodosum, respectively, as well as several minor ones. The 2-DE demonstrated a total of 56 and 41 protein spots for S. nigrogilvum and S. nodosum, respectively. Most of the proteins obtained in both species were enzymes involved in blood feeding, including proteases, apyrases, hyaluronidases, aminopeptidase and elastase. The results obtained in this study provided a new body of knowledge for a better understanding on the role of salivary gland proteins in these black fly species in Thailand.

Matched MeSH terms: Peptide Hydrolases
Fulltext Suppression of Staphylococcus aureus biofilm formation and virulence by a benzimidazole derivative, UM-C162

Kong C, Chee CF, Richter K, Thomas N, Abd Rahman N, Nathan S

Sci Rep, 2018 02 09;8(1):2758.
PMID: 29426873 DOI: 10.1038/s41598-018-21141-2

Staphylococcus aureus is a major cause of nosocomial infections and secretes a diverse spectrum of virulence determinants as well as forms biofilm. The emergence of antibiotic-resistant S. aureus highlights the need for alternative forms of therapeutics other than conventional antibiotics. One route to meet this need is screening small molecule derivatives for potential anti-infective activity. Using a previously optimized C. elegans - S. aureus small molecule screen, we identified a benzimidazole derivative, UM-C162, which rescued nematodes from a S. aureus infection. UM-C162 prevented the formation of biofilm in a dose-dependent manner without interfering with bacterial viability. To examine the effect of UM-C162 on the expression of S. aureus virulence genes, a genome-wide transcriptome analysis was performed on UM-C162-treated pathogen. Our data indicated that the genes associated with biofilm formation, particularly those involved in bacterial attachment, were suppressed in UM-C162-treated bacteria. Additionally, a set of genes encoding vital S. aureus virulence factors were also down-regulated in the presence of UM-C162. Further biochemical analysis validated that UM-C162-mediated disruption of S. aureus hemolysins, proteases and clumping factors production. Collectively, our findings propose that UM-C162 is a promising compound that can be further developed as an anti-virulence agent to control S. aureus infections.

Matched MeSH terms: Peptide Hydrolases
Fulltext Molecular docking analysis of Carica papaya Linn constituents as antiviral agent

Radhakrishnan, N., Lam, K. W., Norhaizan, M. E.

International Food Research Journal, 2017;24(4):1819-1825.
MyJurnal

Carica papaya (papaya) fruits are available throughout the world and it is well accepted as food or as a quasi-drug. Aqueous papaya leaves extract have been used as treatment for dengue fever. This prompted us to carry out the docking study on these nine selected ligands (phyto-constituents of papaya) which are carpaine, dehydrocarpaine I and II, cardenolide, p-coumaric acid, chlorogenic acid, caricaxanthin, violaxanthin and zeaxanthin. These phytoconstituents were evaluated on the docking behaviour of dengue serotype 3 RNA-dependent RNA polymerase (RdRp); influenza A (H1N9) virus neuraminidase (NA); chikungunya virus glycoprotein (E3-E2-E1) and chikungunya virus non-structural protein2 (nsP2) protease using Discovery Studio Version 3.1. In addition, molecular physicochemical, drug-likeness, ADMET (Absorption, Distribution, Metabolism, Excretion and Toxicity) and TOPKAT (Toxicity Prediction by Komputer Assisted Technology) analyses were done. The molecular physicochemical analysis revealed that cardenolide and p-coumaric acid (2 ligands) complied with Lipinski’s rule of five. Dehydrocarpaine II, cardenolide, caricaxanthin, violaxanthin and zeaxanthin all the five ligands were predicted to have plasma protein binding (PPB) effect. Docking studies and binding free energy calculations revealed that p-coumaric acid exhibited very least binding energy irrespective of its target protein. Hence, the results of this present study exhibited the potential of these nine ligands as antiviral agent.

Matched MeSH terms: Peptide Hydrolases
Fulltext Identifications of drug resistance mutations among antiretroviral treatment naive HIV-1 patients in Peninsular Malaysia

Zain RM, Ibrahim N, Ismail S, Suppiah J, Mat Rahim NA, Thayan R

Asian Pac J Trop Med, 2017 Jan;10(1):75-78.
PMID: 28107870 DOI: 10.1016/j.apjtm.2016.12.005

OBJECTIVE: To determine drug resistance mutations and the HIV-1 subtypes among antiretroviral treatment naive HIV-1 patients in Peninsular Malaysia.
METHODS: A total of 45 samples from four hospitals that provide HIV viral load services were subjected to the amplification of the protease and two third of reverse transcriptase regions of the pol gene by RT-PCR and Sanger sequencing. Drug resistance mutation (DRM) interpretation reports the presence of mutations related to protease inhibitors (PIs), Nucleoside reverse-transcriptase inhibitors (NRTI) and Non-nucleoside reverse-transcriptase inhibitors (NNRTI) based on analysis using Stanford HIV database program.
RESULTS: DRMs were identified in 35% of patients, among which 46.7% of them showed minor resistance to protease inhibitor with A71V and L10l were the commonest DRMs detected. About 21.4% and 50.0% of patients had mutations to NRTIs and NNRTIs, respectively. CRF01_AE was found to be the predominant HIV-1 subtype.
CONCLUSIONS: These findings have served as an initial crucial data in determining the prevalence of transmitted HIV-1 drug resistance for the country. However, more samples from various parts of the country need to be accumulated and analyzed to provide overall HIV-1 drug resistance in the country.

Matched MeSH terms: Peptide Hydrolases
Fulltext Inhibition of SARS-CoV-2 3CL protease by the anti-viral chimeric protein RetroMAD1

Chan LC, Mat Yassim AS, Ahmad Fuaad AAH, Leow TC, Sabri S, Radin Yahaya RS, et al.

Sci Rep, 2023 Nov 17;13(1):20178.
PMID: 37978223 DOI: 10.1038/s41598-023-47511-z

COVID-19 results from SARS-CoV-2, which mutates frequently, challenging current treatments. Therefore, it is critical to develop new therapeutic drugs against this disease. This study explores the interaction between SARS-CoV-2 3CLpro and RetroMAD1, a well-characterized coronavirus protein and potential drug target, using in-silico methods. The analysis through the HDOCK server showed stable complex formation with a binding energy of -12.3, the lowest among reference drugs. The RetroMAD1-3CLpro complex underwent a 100 ns molecular dynamics simulation (MDS) in an explicit solvation system, generating various trajectories, including RMSD, RMSF, hydrogen bonding, radius of gyration, and ligand binding energy. MDS results confirmed intact interactions within the RetroMAD1-3CLpro complex during simulations. In vitro experiments validated RetroMAD1's ability to inhibit 3CLpro enzyme activity and prevent SARS-CoV-2 infection in human bronchial cells. RetroMAD1 exhibited antiviral efficacy comparable to Remdesivir without cytotoxicity at effective concentrations. These results suggest RetroMAD1 as a potential drug candidate against SARS-CoV-2, warranting further in vivo and clinical studies to assess its efficiency.

Matched MeSH terms: Peptide Hydrolases
Down-regulation of LPA receptor 5 contributes to aberrant LPA signalling in EBV-associated nasopharyngeal carcinoma

Yap LF, Velapasamy S, Lee HM, Thavaraj S, Rajadurai P, Wei W, et al.

J Pathol, 2015 Feb;235(3):456-65.
PMID: 25294670 DOI: 10.1002/path.4460

Undifferentiated nasopharyngeal carcinoma (NPC) is a highly metastatic disease that is consistently associated with Epstein-Barr virus (EBV) infection. In this study, we have investigated the contribution of lysophosphatidic acid (LPA) signalling to the pathogenesis of NPC. Here we demonstrate two distinct functional roles for LPA in NPC. First, we show that LPA enhances the migration of NPC cells and second, that it can inhibit the activity of EBV-specific cytotoxic T cells. Focusing on the first of these phenotypes, we show that one of the LPA receptors, LPA receptor 5 (LPAR5), is down-regulated in primary NPC tissues and that this down-regulation promotes the LPA-induced migration of NPC cell lines. Furthermore, we found that EBV infection or ectopic expression of the EBV-encoded LMP2A was sufficient to down-regulate LPAR5 in NPC cell lines. Our data point to a central role for EBV in mediating the oncogenic effects of LPA in NPC and identify LPA signalling as a potential therapeutic target in this disease.

Matched MeSH terms: Phosphoric Diester Hydrolases/physiology

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