Malaysian Journal of Biochemistry
& Molecular Biology
(E-ISSN: 2600-9005)
The Official Publication of the Malaysian Society for Biochemistry & Molecular Biology (MSBMB)
Indexed by SCOPUS and Malaysian Citation Index (MYCITE)
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December 2022
Malay. J. Biochem. Mol. Biol. (2022) 25 (3)
TABLE OF CONTENTS
Page 1- 8
Ikrimatul Hasnah, Wira Eka Putra, Sustiprijatno, Arief Hidayatullah, Dahlia Normala, Nurrasilfa, Rani Dwi Lestari, Siti Nurazizah, Yusniar Zurroh Asfiniya, Diana Widiastuti, Hendra Susanto, Muhammad Fikri Heikal, Hary Isnanto
IN SILICO STUDY REVEALED THE ANTI-VIRAL POTENCY OF Allium cepa BIOACTIVE COMPOUNDS INHIBIT M-PRO SARS-COV-2 PROTEIN
Abstract
A zoonotic disease called COVID-19 was initially spread from animals to people. SARS-CoV-2, as opposed to SARS, was a pandemic that resulted in about 274,676,729 cases globally. Numerous studies have looked into potential novel treatments for COVID-19 infection. In this study, 16 phytochemicals from Allium cepa L. were examined in silico for their potential to bind to the primary protease of COVID-19 (PDB ID: 6LU7). Lipinski's rules are used to choose the ligands. Protein protease's (Mpro) Cys-145 and His-41 are its active sites. The chosen ligands are examined using molecular docking with PyRx and Vina Wizard and 2D visualization with LigPlot+. Selected ligands' binding energy value will be contrasted with hydroxychloroquine as a control. According to the results, the compounds luteolin (CID: 5280445), isorhamnetin (CID: 5281654), and apigenin (CID: 5280443), which had binding energies of 7.4, 7.2, and -7 kcal/mol, respectively, become potential COVID-19 inhibitors. These substances have higher binding energies than the control, hydroxychloroquine (CID: 3652), which has a lower binding energy (-6 kcal/mol). Due to its important pharmacokinetic features, luteolin demonstrated the best binding efficacy to Mpro, enabling the development of new drugs.
Page 9 - 17
Kingsley Ikechukwu Ijoma, Vincent Ishmael Egbulefu Ajiwe, Juliana Onyema Ndubuisi
EVIDENCE-BASED PREFERENTIAL IN VITRO ANTISICKLING MECHANISM OF THREE NATIVE NIGERIAN PLANTS USED IN THE MANAGEMENT OF SICKLE CELL DISEASE
Abstract
Sickle cell anemia is a hereditary genetic disease caused by the substitution of glutamic acid by valine at beta six (β-6) position of the hemoglobin; the clinical implication is that the erythrocyte contains the hemoglobin polymerizes, leading to severe clinical consequences. Therefore, we investigated the antisickling mechanism of Ficus thonningii (FTH), Jatropha tanjorensis (JTR) and Justicia carnae (JCN) native to Southeast Nigeria used in the management of sickle cell disease. The experiment was designed to include erythrocyte fragility, erythrocyte reversibility and polymerization inhibition mechanisms. The results obtained were analyzed using one-way ANOVA and Tukey Posthoc test. From the results of the erythrocyte fragility test, it was observed that at 0.35% saline concentration, FTH, JTR and JCN extracts reduced hemolysis to 26.548±0.056%, 18.055±0.064%, and 20.217±0.035% respectively while hydroxyurea (control drug) reduced hemolysis to 14.459±0.040%. In the presence of the control, the percentage number of sickle cells was 91.001±0.170% whereas, hydroxyurea was 30.414±0.162% while, extracts of FTH, JTR and JCN reduced sickling to 49.818±0.082%, 41.001±0.413% and 33.957±0.062% respectively. Also, the results of the polymerization inhibition analysis showed that extracts of FTH, JTR and JCN had relative percentage polymerization inhibition of 76.888±0.042%, 48.723±0.113% and 75.447±0.063%, respectively, in comparison to hydroxyurea which inhibited sickle erythrocyte polymerization by 70.903±0.150%. Our findings suggest the preferential but variable antisickling mechanism of the studied extracts; hence, the leaves of the assayed plants contain potential antisickling phytochemicals and should be explored further for their antisickling benefits.
Page 18-26
Nurul Syafika Muslimin, Farida Zuraina Mohd Yusof, Nur Qursyna Boll Kassim, Khalilah Abdul Khalil
MOLECULAR IDENTIFICATION OF FUNGI ASSOCIATED WITH PLANT ROT DISEASE OF SOURSOP (Annona muricata) IN SERI MENANTI, NEGERI SEMBILAN
Abstract
Soursop (Annona muricata) is renowned for its ethnomedicinal uses, leading to the increasing demand for this fruit. Soursop rot disease has become one of the major constraints in soursop fruit production in Peninsular Malaysia, especially for the smaller commercial producer, much likely due to the indefinite identity of pathogens associated with the disease. As such, this study was designed as a preliminary identification of the fungal species that contribute towards soursop rot diseases in Negeri Sembilan, Malaysia using the ITS primer. The infected soursop leaves and soil samples were collected from a farm located in Seri Menanti, Negeri Sembilan, Malaysia. Cultivation was done on Potato Dextrose Agar (PDA) medium. Twenty-five (25) isolates were collected, and six (6) different species of fungi were discovered through molecular identification. They were suggested to have different roles and contributions in various types of crops worldwide, namely the Gongronella butleri, Fusarium beomiforme, Lasiodiplodia theobromae, Hypocreals sp., Neopestalotiopsis sp. and Colletotrichum siamense.
Page 27-36
Puteri Nur Farahin, Normah Haron, Deny Susanti, and Noor Hasniza Md Zin
Anacardium occidentale Linn. LEAVES AND ITS MEDICINAL PROPERTIES
Abstract
Anacardium occidentale leaves, known as cashew leaves or Pucuk Gajus in Malay, is a member of the Anacardiaceae family that is widely grown in tropical countries such as Malaysia, India and Brazil. It had been traditionally used for treating many health problems as it contained many beneficial phytochemical compositions such as polyphenols, flavonoids, tannin, vitamin C and carotenoids. Its extract possesses antidiabetic activity, which prevents a sudden rise in postprandial blood glucose level by inhibiting α-amylase to slow down the breakdown of long-chain carbohydrates to glucose. It also controls the blood glucose level by inhibiting dipeptidyl peptidase IV (DPPIV), which prevents the rapid degradation of incretin and thus stimulates insulin secretion. Besides, it acts as an antioxidant agent to protect against lipid peroxidation and scavenge radicals. It showed antimicrobial properties by inhibiting the growth of pathogenic bacteria, including Gram-positive Staphylococcus aureus and Bacillus subtilis, and Gram-negative Escherichia coli and Klebsiella pneumonia, and fungi which are Aspergillus niger, Penicillium digitatum and Colletotrichum gloeosporiodes. Furthermore, it exhibited an antiulcerogenic effect by preventing gastric lesions and anti-inflammatory properties, which interrupt the inflammation process by controlling cytokines secretion in macrophage lipopolysaccharide-stimulated cells. Therefore, A. occidentale may potentially be contributed to the treatment of various diseases, but continuous study is needed to determine further the bioactive compounds that are responsible for the mechanism of action.
Page 37-46
Semaa A. Shaban, Hanan Mohammed Saied, Safa Sadeq Fayez, Ahemd AbdulJabbar Suleiman
STUDY OF THE EFFECT OF DIFFERENT miRNA ON GENE EXPRESSION PROFILE DURING Mycobacterium tuberculosis INFECTION USING BIOINFORMATICS TOOLS
Abstract
Tuberculosis (TB) is one of the most common diseases worldwide. Mycobacterium tuberculosis (Mtb) is the main causal agent of tuberculosis. Changes in gene expression are very important for normal biological functions. Changes in gene expression have adverse effects and cause many diseases. Tb is one of them. Gene expression is controlled by many small noncoding miRNAs. In this study, we identified relationships between miRNAs and differentially expressed genes in tuberculosis by using bioinformatics tools. Dataset GSE51029 was downloaded from the GEO database to check for expression. DEGs in the microarray dataset were identified by a bioinformatics tool, namely GEO2R. Enrichment analysis of DEGs was done through DAVID. The PPI network was constructed through the Cytoscape of the DEGs. Hub-genes were identified by using cyto-Hubba extracted from the PPI network. The gene’s interaction network of miRNA-hub was constructed to evaluate the miRNAs interacting with hub genes. A total of 285 DEGs were identified. Of those, 82 genes were downregulated and 202 genes were upregulated. Enrichment analysis of DEGs showed their involvement in cell surface receptors, regulation of proteins, signaling pathways, cytoplasmic endoplasmic reticulum, endoplasmic, nuclear membrane, transcription coactivator activity, mRNA binding, protein kinase binding, and deacetylase binding. KEGG analysis presented that identified DEGs were involved in signaling pathways and other diseases. The ten genes closely related to miRNA have also been identified. This study shows that identified DEGs, pathways, and miRNAs that closely interact with hub genes are involved in causing tuberculosis. Their clinical evaluation at an early stage may provide important targets for the treatment of tuberculosis.
Page 47-57
Siti Norazura Jamal, Dynna Ayun Donny and Dhilia Udie Lamasudina
THE INFLUENCE OF ENZYMATIC HYDROLYSIS ON ANTIMICROBIAL ACTIVITY AGAINST RICE PATHOGENS FROM Bactronophorus thoracites (SHIPWORM) PROTEIN HYDROLYSATE
Abstract
The outbreak of leaf blight disease in Malaysia affects the sustainability of rice production in supporting the growing population. Bactronophorus thoracites is a wood-boring shipworm with high protein and possesses numerous bioactive compounds. The present study investigated the effects of enzymatic hydrolysis for generating molluscs protein hydrolysates (MPH) from B. thoracites and their efficacy against rice pathogens. The inhibition percentage against rice pathogens was significantly (p<0.05) increased when enzyme concentration was increased from 1.75% to 2.05% (w/v) but slightly decreased at a concentration exceeding 2.35%. The inhibition percentage was also found to increase when the temperature increased to 55°C. However, the hydrolysis at 65°C produced a lower inhibition percentage. Prolonging the incubation time from 120 to 240 minutes significantly (p<0.05) increased the inhibition percentage. The inhibition percentage of hydrolysates was significantly (p<0.05) increased when pH increased from 9.0 to 9.5. However, increasing pH to 10.0 resulted in a reduction in inhibition percentage. Subsequently, the MPH with optimum condition (2.05% w/v, 55°C, 240 min and pH 9.5) was lyophilised and analysed for antimicrobial activities. The MIC and MBC values for MPH were 500 μg/mL and 1000 μg/mL against P. ananatis and 250 μg/mL and 1000 μg/mL against P. stewartii. The zones of inhibition value for MPH were 20.76 mm (1000 μg/mL) and 11.53 mm (500 μg/mL) for P. ananatis and 21.1 mm (1000 μg/mL), 14.03 mm (500 μg/mL) and 8.16 mm (250 μg/mL) for P. stewartii. At MIC, 2 MIC, and 4 MIC, the MPH exhibited a longer lag phase. Nonetheless, at a lower MIC concentration, a progressive rise in bacterial growth for P. ananatis and P. stewartii at 12 hours was observed. This study shows that enzymatic hydrolysis using alcalase can produce potential antimicrobial agent applicable to the agricultural sector.
Page 58-64
Wan Xin Goh, Jun Wei Chiang, Nyet Kui Wong, Soi Moi Chye
Moringa oleifera SEED EXTRACT INHIBITS OXIDATIVE STRESS AND CYTOTOXICITY VIA INTRINSIC AND EXTRINSIC PATHWAYS IN HIGH GLUCOSE-INDUCED RINM5F β-CELLS
Abstract
Hyperglycemia is one of the hallmarks of diabetes mellitus and is known to cause apoptosis in pancreatic islets. A reduction in pancreatic β-cell number and function results in impaired insulin production and diabetes. Moringa oleifera has substantial evidence of anti-diabetic and anti-hyperglycemic activity. However, there is a lack of study on the anti-cytotoxicity mechanism of Moringa oleifera seed extract in high glucose-induced RINm5F β-cells. In this study, we investigate the effect of Moringa oleifera seed extract on cytotoxicity, oxidative stress, and caspases pathway in high glucose-induced RINm5F β-cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay results showed that Moringa oleifera seed extract inhibits high glucose-induced cytotoxicity in RINm5F β-cells. Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay demonstrated that Moringa oleifera seed extract inhibits oxidative stress in high glucose-induced RINm5F β-cells. Moreover, Moringa oleifera seed extract inhibits high glucose-induced cytotoxicity via intrinsic and extrinsic pathways in RINm5F β-cells.
August 2022
Malay. J. Biochem. Mol. Biol. (2022) 25 (2)
TABLE OF CONTENTS
Page 1- 11
Afiqah Insyirah Lutfi, Nantacha Kritsanayanyong, Siti Nur Diyanah Rahim, Noor Baity Saidi, Mohd. Puad Abdullah and Nadiya Akmal Baharum
FUNCTIONAL PREDICTION OF PATHOGENESIS-RELATED 10 IN Musa acuminata DH PAHANG (MaPR-10) FOR TARGETED BANANA ADAPTATION AGAINST STRESSES
Abstract
Pathogenesis-related 10 (PR-10) is a defense-related protein that provides adaptive responses to various biotic and abiotic stimuli. It was characterized by diverse roles due to its multigene property. Hence, the specific biological roles of PR-10 are still inconclusive. To date, there is no reported in-silico characterization of PR-10 in wild bananas yet. Thus, this study predicts the valuable Musa acuminata DH Pahang (wild banana) MaPR-10 copies which might be useful for the targeted functions. A total of ten putative MaPR-10 members were discovered which can be clustered into three major subgroups (intracellular PR-10 (IPR), S-norcoclaurine synthase (NCS) and major latex protein (MLP) through phylogenetic analysis. MEME suite analysis displayed distinguished motif arrangement among all three subgroups. Gene ontology (GO) revealed that all MaPR-10s exhibit similar functions such as plant defence, phosphoprotein phosphatase activity and activation of ABA signaling pathway except for Ma09_p15840.1 (an IPR) and Ma04_p33910.1 (NCS) which displayed distinct roles both in biological and molecular functional prediction. Furthermore, analysis of the promoter regions using PLACE presented a diverse regulation of MaPR-10s upon various biotic and abiotic stimuli. Altogether, this study contributes to a better perspective of MaPR-10s features and their functional predictions, which will be essential for future crop improvements against biotic and abiotic stresses, particularly in bananas.
Page 12 - 27
Nik Yusnoraini Yusof, Muggunna Balasubramaniam, Anis Fathima Abdul Kareem and Amira Suriaty binti Yaakop
COMPARATIVE GENOMICS OF Xanthomonas BACTERIA: INSIGHTS INTO ITS PLANT PATHOGENICITY
Abstract
Xanthomonas is a group of Gram-negative bacteria from class Gammaproteobacteria that causes multiple diseases in many plant hosts. Bacterial secretion systems of Xanthomonas contribute to its pathogenicity and are one of the most important factors for symptom and disease development. Comparative genomic analysis between 20 Xanthomonas bacteria, well known for their pathogenicity was chosen to study the bacterial secretion system, specifically, the type III bacterial secretion system associated with bacterial pathogenesis. The analysis was performed using databases and software including NCBI, RAST, antiSMASH, KEGG and Islandviewer4 to compare the biological and taxonomical relationship between these genomes. The average nucleotide identity (ANI) similarity index ranges between 79% to 99.96%, indicating that the selected genomes were closely related. Screening of secondary metabolites using antiSMASH showed all 20 genomes produced secondary metabolites. Siderophore secondary metabolite gene cluster necessary for optimum virulence was present in all the genomes. Genomic island discovery using Islandviewer4 revealed the presence of mobile genetic elements such as phages and transposons, indicating horizontal gene transfer events. Genes and pathogenicity pathway mechanism identification of type III bacterial secretion system was done using SEED subsystem and KEGG database. A total of 34 genes associated with type III bacterial secretion system were identified. HrpA, HrpB, HrpX and HrpG were the conserved genes across the Xanthomonas species. The HrpX and HrpG genes are essential for symptom development, disease establishment and invasion into host cells.
Page 28-36
Anh Cam Ha, Tan Minh Le, Chinh Duc Nguyen Pham
FLAVONOIDS-ENRICHED EXTRACTION OF Phyllanthus amarus LEAVES AS A POTENTIAL IN VITRO MEDICATION FOR TREATING DIABETES
Abstract
This study aimed to evaluate the extraction process of Phyllanthus amarus for the enrichment of flavonoid content applied in treating diabetes and its complications. The investigated factors included ethanol concentration (0, 30, 70, 100%), extraction temperatures (40, 50, 60, 70 °C), extraction time (30, 60, 120 min), and solid-to-solvent ratio (1:10, 1:15, and 1:20 g:mL). The results showed that the appropriate extraction conditions for high flavonoid content were at an ethanol concentration of 70%, extracting temperature of 50 oC, extraction time of 60 min, and solid-to-solvent ratio of 1:15 g:mL. The relevance of flavonoid and α-glucosidase inhibition activity was also confirmed in this study. This enriched-flavonoid extract exhibited good 𝛼𝛼-glucosidase inhibition activity with a half-maximal inhibitory concentration (IC50) of 2 μg/mL, antioxidant activity with IC50 of 4.5 μg/mL, and anti-inflammatory activity with IC20 of 400 μg/mL. Besides, the extracts also displayed activities in antimicrobial, especially Gram-positive strains.
Page 37-42
Boon Hong Kong, Rumiyati Rumiyati, Muhammad Fazril Mohamad Razif, Shin Yee Fung
EVALUATION OF CHEMICAL COMPOUNDS FOUND IN Volvariella volvacea FROM RURAL SEDAYU, YOGYAKARTA, INDONESIA
Abstract
The Volvariella volvacea or commonly known as the straw mushroom is one of the nutrient-rich mushrooms. V. volvacea mushroom collected from Sedayu, Yogyakarta, Indonesia was analyzed for its protein, carbohydrate, and glycoprotein contents. The proteins isolated from the cold water extract of V. volvacea were identified using liquid chromatography-tandem mass spectrometry. Results showed that the cold water extract of V. volvacea contained moderate amounts of proteins and carbohydrates, and was rich in glycoproteins. A pore-forming cardiotoxic protein, Volvatoxin A2, was identified in the V. volvacea; however, the involvement of this toxin as the causative agent in poisoning from V. volvacea consumption remains to be elucidated.
Page 43-51
Derouiche Samir, Ahmouda Ibtissam, Moussaoui Rokaya
STUDY OF THE TOXIC EFFECT OF DRINKING WATER CONTAMINATED WITH LEAD ON LIVER AND KIDNEY FUNCTIONS IN PREGNANT RATS: THE BENEFIC ROLE OF Aquilaria malaccensis L. SPICE
Abstract
The objective of this work was to study the effect of using Aquilaria malaccesis spice on some biological parameters in pregnant rats exposed to lead-contaminated drinking water. Twenty (20) female rats were randomly divided into four groups (n = 5): control, pregnant rats, pregnant + Pb and pregnant +Pb+ A. malaccensis (Aq). Lead (200 mg/kg b.w.) as Pb (C2H3O2)2 was added to their drinking water for 20 days. A. malaccensis (heartwood powder at a dose of 10 g/kg of diet) was added to the feed during the last 10 days of lead exposure in the animals. The results showed that A. malaccensis was rich in various flavonoid and phenolic compounds. On the other hand, the results showed that pregnancy caused significant physiological and biochemical changes in rats. Our results also showed a significant increase (p <0.05) of liver lead, blood glucose, lipids profile, serum uric acid, urea, transaminases, MCV, Platelet, MDA and GST levels in the pregnant rats Pb2+group compared to the pregnant rats. The results obtained also revealed a significant decrease (p <0.05) in the level of RBC, WBC, Hb, hematocrit, GSH and SOD activity in comparison with pregnant rats group. The treatment with A. malaccensis spice partially improved the biochemical and hematological parameters, which protected the tissues against radical attacks (oxidative stress) caused by lead in pregnant rats. In conclusion, this work shows that A. malaccensis spice has beneficial effects in reducing oxidative stress and toxic effects of lead on the liver and kidneys in rats during gestation.
Page 52-59
Hannah Zaihal Hazri, Nurfatini Zakaria, Muhajir Hamid, Mohd Badrin Hanizam Abdul Rahim and Mohd Ezuan Khayat
SCREENING OF DRUG LEAD CANDIDATES FROM Eurycoma longifilia AND
Crocus sativus FOR DPP-4 INHIBITION VIA IN SILICO PHARMACOKINETIC AND MOLECULAR DOCKING ANALYSES
Abstract
Dipeptidyl peptidase 4 (DPP-4) is an enzyme responsible for the inactivation of incretion hormones, and its inhibition augments insulin release from the pancreas. Over the past years, the inhibition of DPP-4 has emerged as an important therapeutic route to treat type 2 diabetes (T2D) since it offers a lower risk of hypoglycemia. Currently, there are several DPP-4 inhibitors available in different molecular structures. The structural diversity of these inhibitors proves the flexibility of the enzyme to bind various shapes of inhibitors. This opens the opportunity for more discovery of novel DPP-4 inhibitors. In this study, 15 phytocompounds from Eurycoma longifilia and Crocus sativus were screened to obtain the candidate leads for DPP-4 inhibitor through in silico pharmacokinetics and molecular docking studies. In order to assess the drug-likeness of the phytocompounds, Lipinski rule of 5 was applied. Thirteen out of 15 phytochemicals were found to pass all the criteria set for this rule. Next, the absorption, distribution, metabolism, and excretion – toxicity (ADMET) profile of the 13 compounds were studied and the result showed only four of them (eurycomalactone, crocetin, picrocrocin and crocusatin D) possessed the characteristics of the good drug. Via molecular docking, it was found all four compounds bound to DPP-4, with the highest binding affinity was observed in eurycomalactone-DPP4 interaction. Eurycomalactone was found to bind to DPP-4 at His363, Ile407, Ile374, Arg356, as well as two amino acids that have previously shown to interact with DPP-4 inhibitor, Phe357 and Arg358. This study indicates that eurycomalactone from Eurycoma longifilia can potentially be developed into a new dpp-4 inhibitor.
Page 60-68
Lamiae Amallah, Ismail El Moudden, Fatima Gaboun, Mohan D Pant, Souad Skalli, Rachida Hassikou and Mona Taghouti
ASSESSMENT OF THE EVOLUTION OF DURUM WHEAT END-USE QUALITY FROM DOMESTICATION TO BREEDING ACHIEVEMENTS
Abstract
The quality characteristics of durum wheat are important parameters to consider in developing grain products. Here we assessed the end-use quality of a collection of 294 durum wheat from several genetic resources including Mediterranean landraces, Moroccan varieties, and international lines using various standardized methods. To achieve this goal, sample seeds for each genotype were subjected to the most important features determining the commercial value of durum wheat, according to standardized methods; gluten strength (SDS), grain protein content (PC), yellow pigment concentration (YP), yellow index (b), brightness (L), test weight (TW), 1000-kernel weight (TKW), and kernel vitreousness (VIT) were assessed. There were decreases over time in yellow pigment content and protein levels from landraces to Moroccan cultivars, via international lines. These findings might be directly used by farmers interested in cultivating traditional varieties for specialized food markets. Genotype selection based on multiple traits is a key issue in plant breeding; breeders practice selection on target traits to improve productivity but need to consider unfavorable associations among key traits of similar economic interest.
Page 69-76
Lucky Poh Wah Goh, Arnold Marshall Molujin, Roslina Jawan, Mohd Khalizan Sabullah, Suraya Abdul Sani and Jualang Azlan Gansau
ANTI-CANCER POTENTIAL OF STING AND STINGLESS BEE PRODUCTS: AN UPDATED SYSTEMATIC REVIEW
Abstract
Modern science and tools have found that traditionally used bee products could serve as alternative medicine and treatments. However, bee is under-studied and the bioactivity of sting and stingless bee products remain inconclusive. This systematic review aims to report a concise and updated finding on the potential anti-cancer activities of bee products. The article search strategy was performed according to PRISMA guide-lines and retrieved 425 articles. The authors assessed the titles, abstracts and full-text articles for suitability and data retrieval. The data from 16 articles were deliberated and summarized. Based on the information of this systematic review, bee products have great anti-cancer activity against various types of cancers and the activity is dependent on the type of product bee species. Bee species products can be further utilized and explored as an alternative medicine for cancer patients' treatments or healthy individuals as preventive medicine.
Page 77-85
M.Z. Nawahwi, S. N. Maadon, N.A Hasan, N.I Hasbullah, M.S Mohd Zan
ISOLATION AND IDENTIFICATION OF ENDOPHYTIC FUNGI FROM UiTM-PERHILITAN RESEARCH STATION, KUALA KENIAM, PAHANG, MALAYSIA
Abstract
Endophytic fungi live inside plant tissues and do not degrade or produce chemicals that infect the host cell. Because of their species richness and diversity, endophytic fungi are an underutilised source of pharmacologically relevant chemicals. Hence, this study was conducted to isolate, identify and characterize endophytic fungi from the sampling site at UiTM-Perhilitan Research Station in Kuala Keniam, Pahang, Malaysia. In the present work, three different components (leaf, root, and stem) of four different plant species were randomly collected. The morphology of isolates was characterized based on mycelial texture, pigmentation, colony appearance, and spores, which were then subjected to molecular analysis. In total, 32 endophytic isolates were successfully obtained. Relatively, leaf samples were found to have greater colonization frequency in plant B (72.7%) as compared to root and stem parts. Morphological analysis clustered the isolates into 15 different groups and revealed that all the isolates belonged to Ascomycota and Basidiomycota phylum. Molecular identification by using ITS universal primer identified two isolates as Candida metapsilosis (C9) and Clonostachys roses (D13). However, 12 isolates were recorded as unidentified groups. The findings of this study point to the possibility of further research on bioactive substances produced by these two endophytic fungi.
Page 86-91
Mohammed Auwal Ibrahim, Auwal Adamu, Murtala Bindawa Isah, Aminu Usman, Raphael Aminu, Mujaheed Abubakar, Hassan Ibrahim, Khadija Aminu, Fauziyya Lawal Ahmad
STIGMASTEROL ATTENUATES OXIDATIVE STRESS IN SOME ORGANS OF Trypanosoma congolense INFECTED RATS
Abstract
Stigmasterol has shown positive effects against Trypanosoma congolense, which induced organ pathological changes in rats. Therefore, to unravel a possible mechanism of this action, the effect of stigmasterol on oxidative stress in the liver, kidney, spleen, and heart of T. congolense infected rats was investigated. Rats were infected with T. congolense and on day 11 post-infection, the infected rats were treated with 100 and 200 mg/kg body weight (BW) of stigmasterol for 14 days. At the end of the experiment, the levels of reduced glutathione, malondialdehyde as well as the activities of catalase and superoxide dismutase were analyzed in the homogenates of liver, kidney, spleen, and heart obtained from the stigmasterol-treated infected rats and compared to non-infected and infected non-treated controls. The data showed evidence of induction of oxidative stress in all the organs caused by the infection. Treatments with the 100 and 200 mg/kg BW of stigmasterol attenuated the T. congolense induced oxidative stress in all the organs as indicated by significant (P < 0.05) amelioration of the levels of most of the markers. A similar trend was observed with diminazene aceturate, used as a standard drug. It is concluded that stigmasterol could attenuate T. congolense induced oxidative stress which might be due to a direct free radical scavenging effect or a consequence of the reported parasite clearance.
Page 92-102
Nor Sabrina Shek Daud, Anna Pick Kiong Ling, Ying Pei Wong, Rhun Yian Koh and Kenny Gah Leong Voon
ROLE OF ORIENTIN AGAINST BETA-AMYLOID INDUCED CELL DEATH IN SH-SY5Y HUMAN NEURONAL CELLS
Abstract
Alzheimer’s disease (AD) is the most common form of dementia. Accumulation of beta-amyloid (Aβ) plaque has led to anti-inflammatory response, tau hyperphosphorylation and eventually neuronal apoptosis. Current treatment options only reduce the symptoms of AD but not curing the root cause of the problem. This brings us to a search for potential drugs that could tackle the cause of AD. Past studies have shown that orientin is neuroprotective due to its antiapoptotic, anti-inflammatory and antioxidative properties. Hence, this study aims to investigate the mechanism of action of orientin against Aβ-induced cell death in SH-SY5Y human neuronal cells. The SH-SY5Y cells were pre-treated with maximum non-toxic dose (MNTD) and half MNTD (½ MNTD) of orientin (20 μM and 10 μM, respectively) for 4 hours and further exposed to the inhibitory concentration of 50% (IC50) of Aβ (53 μM) for 24 hours. After pre-treating the Aβ-challenged cells with orientin, the cell apoptosis, caspase 3/7, 8 and 9 assays as well as the expression of mitogen‑activated protein kinase (MAPK) proteins were determined. MNTD, ½ MNTD and IC50 were pre-determined using 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Cell apoptosis was evaluated by Hoechst staining and changes in caspases were determined using luminescent assay kits. Finally, protein detection was carried out using western blot. The percentage of apoptotic cells exposed to Aβ was reduced when pre-treated with orientin either at MNTD or ½ MNTD. Findings from caspase activities revealed a reduction in caspase 3/7 activities when pre-treated with orientin at MNTD. Analysis of the MAPK pathway revealed that orientin at ½ MNTD could downregulate the expression of p38 and JNK proteins. This study demonstrated that orientin could inhibit Aβ-induced cell death in SH-SY5Y cells through the MAPK signalling pathway and caspase cascade.
Page 103-113
Nur Suraya Ashikin Rosli, Shafinaz Abd Gani, Mohd Ezuan Khayat, Uswatun Hasanah Zaidan and Mohd Badrin Hanizam Abdul Rahim
COMPARING THE BEHAVIOUR OF HUMAN AND RODENT BETA CELL LINES FOR IN VITRO DIABETES MELLITUS STUDY
Abstract
For the past 30 years, considerable effort has been made to generate useful rodent and human insulin-secreting cell lines to understand the complexity and functionality of the pancreatic beta cells. Since then, numerous types of research have been carried out using these cell lines to assess the pathology of diabetes mellitus. Among them, the cell lines INS-1, MIN6, 1.1B4 and EndoC-βH1 are the most widely used due to their stability and appropriate response to glucose stimulation. With multiple choices of cell lines available, there is a question of which cell line can be the best cell model for in vitro pancreatic beta cell study. The characterisation of each of these cell lines has already been vigorously studied, but there is still the need to understand how rodent and human cell lines differ from each other in order to choose the most appropriate cell model for a specific study. In this review, we explore the differences between human and rodent insulin-secreting cell lines in terms of their culturing condition, glucose-stimulated insulin secretion and reaction toward oxidative stress. Together, it is hoped that these comparisons could provide new perspectives for researchers in dealing with the available insulin-secreting cell lines for their in vitro diabetic studies.
April 2022
Malay. J. Biochem. Mol. Biol. (2022) 25 (1)
TABLE OF CONTENTS
Page 1- 7
Huynh Nguyen Que Anh, Le Pham Tan Quoc
MICROWAVE-ASSISTED EXTRACTION OF PHENOLIC COMPOUNDS FROM
Polyscias fruticosa (L.) Harms ROOT
Abstract
Polyscias fruticosa (L.) Harms is an herbal plant that possesses many medicinal uses. The interest in this material and its applications have been increasing rapidly in recent years. However, changes in phytochemical components and antioxidant properties of P. fruticosa roots when using microwave-assisted extraction (MAE) methods have not been well understood. The aim of this study was to determine the best extraction conditions for total phenolic content (TPC) and antioxidant capacity (AC) of P. fruticosa roots using MAE method. Four factors of the extraction process (solvent/material (SM) ratio, solvent concentrations, microwave power, and extraction time) were investigated. DPPH method was used to determine free radical scavenging activities while TPC was estimated by the Folin-Ciocalteu assay. The results pointed to the optimal extraction parameters which were ethanol concentration of 50% (v/v), ethanol/material ratio of 60/1 (mL/g), microwave power of 265 W, and extraction time of 5 min. TPC and AC obtained were approximately 2.31±0.01 mg GAE/g DW and 76.62±0.23%. In addition, the initial material was completely destroyed under microwave treatment. The results also indicated that MAE could be a fast and reliable method for quantitative analysis of phenolic compounds in P. fruticosa roots.
Page 8-17
Anatolii Onishchenko, Galina Gubina-Vakulyck, Oleksandr Knigavko, Ketino Sharashydze, Olena Pionova, Dmytro Butov, Hanna Polikarpova, Anton Tkachenko
INTAKE OF SEMI-REFINED CARRAGEENAN CAUSES LOW-GRADE COLONIC INFLAMMATION AND ALTERS EXPRESSION OF EPITHELIAL-MESENCHYMAL TRANSITION MARKERS
Abstract
The aim of this research was to evaluate the effects of semi-refined carrageenan consumed orally on the amount of CD3 and CD68 positive cells residing in the colon and the expression of endothelial-mesenchymal transition (EMT) markers.
Materials and methods. To assess colonic expression of CD3+, CD68+, fascin, vimentin and E-cadherin, sections obtained from 8 rats orally exposed to semi-refined carrageenan (E407a) at a dose of 140 mg / kg of weight during 14 consecutive days and 8 control animals were immunostained with the corresponding antibodies. Levels of expression were assessed quantitatively.
Results. Oral exposure to semi-refined carrageenan resulted in an increase in CD3 and CD68 positive cells in the colonic lamina propria. Quantitative analysis of fascin, vimentin and E-cadherin immunostaining revealed changes in expression of these EMT markers both in the colonic stroma and epithelial cells. Vimentin and fascin were overexpressed in stromal and epithelial cells, while E-cadherin was upregulated in the stroma and downregulated in epithelia.
Conclusions. Our observations suggest that oral intake of semi-refined carrageenan results in the development of low-grade colonic inflammation accompanied by infiltration with CD3 and CD68 positive cells and changes in the expression of EMT markers.
Page 18-27
Maryam Usman Ahmed, Isaac John Umaru and Ismaila Yada Sudi
TOXICOLOGICAL EVALUATION OF TWO FRACTIONS FROM Annona senegalensis
ROOT BARK
Abstract
Annona senegalensis root bark is widely used as a medicinal plant for the treatment of a wide array of diseases. This study evaluated the safety of solvent fractions obtained from its aqueous root bark extract. Crude aqueous extract was partitioned into hexane, dichloromethane and ethylacetate fraction by solvent-solvent fractionation. Albino rats were administered 100, 200 and 400 mg/kg b.wt. Dichloromethane and ethylacetate fractions for 14 days. The effect of their administration on liver, kidney, antioxidant enzymes activities, lipid peroxidation and heamatological parameters was investigated. Both fractions significantly increase alanine transaminase activity, Na+ and K+ concentration, glutathione peroxidase activity and malondialdehyde concentration. They fraction significantly decreased urea concentration and serum superoxide dismutase activity. Dichloromethane fraction significantly increase the PCV while there was no significant difference in all heamatological parameters of rats treated with ethylacetate fraction from aqueous root bark (EFAR) when compared with the control. The two fractions are toxic and should be used with caution.
Page 28-36
Fauziatul Fitriyah, Yora Faramitha, Dini Astika Sari, Irma Kresnawaty
MOLECULAR IDENTIFICATION OF LOCAL CYANOBACTERIUM AND SCREENING FOR BIOHERBICIDE ACTIVITY IN MODEL PLANT
Abstract
Cyanobacteria produce a wide range of secondary metabolites with promising possible applications in agriculture, human health, or industry. This study aims to identify cyanobacterium using the rbcL gene as a molecular barcode and screen for the herbicidal activity of water and ethanol crude extract of the strain. DNA of cyanobacterium strain was extracted with modified CTAB method and then amplified using 1AB_rbcL primers and sent for sequencing. BLAST analysis showed that the strain belongs to Synechocystis sp. The strain was then cultured in 2 L of f/2 culture medium for harvesting. Harvested Synechocystis sp. was extracted in water and 70% ethanol with the ratio of 1:3 (w/v) in an 80°C water bath for 2 hours and ethanol extract was evaporated at room temperature. The herbicidal activity was assayed by germinating sorghum in 0.5% extract with glyphosate (Roundup) as positive control and water as a negative control in 5 replications. Germination rate, shoot, and root length were analyzed statistically with one-way ANOVA in GENSTAT software, Duncan post hoc analysis was performed. The local axenic isolate of cyanobacterium from Indonesia was identified as Synechocystis sp. based on the rbcL gene sequence. The ethanol extract of Synechocystis sp. promotes germination in sorghum seeds compared to water extract, but both were still lower compared to the sorghum seeds germination under glyphosate-based herbicide treatment. The ethanol extract also showed higher root growth inhibition compared to the water extract, but both extracts had no activity on shoot inhibition of sorghum seedling compared to the negative control.
Page 37-50
Mei Wei Lai, Woon Ling Chia, Yee Soon Ling, Yen Min Lan, Sheh May Tam. Chung Cheng Richard Kong, Ming Hock Haw, Yang Mooi Lim
MORPHOLOGICAL CHARACTERISTICS, DNA BARCODING AND METABOLITE PROFILING OF Oldenlandia diffusa (Willd.) Roxb. AND Oldenlandia corymbosa L.
Abstract
Oldenlandia diffusa (Willd.) Roxb. is a well-known traditional Chinese medicinal herb and frequently replaced by Oldenlandia corymbosa L. However, their roles and efficacies are not the same. Therefore, the purpose of this study was to distinguish these two plants based on morphology and DNA barcodes, and to reveal their putative compounds. Wild O. diffusa and O. corymbosa were collected from Johor and Negeri Sembilan in Malaysia, respectively. The morphological characteristics of both plants were recorded during plant collection. DNA barcodes of ribulose-1,5-bisphosphate carboxylase (rbcL), maturase K (matK), and nuclear ribosomal internal transcribed spacer (ITS) were used to identify these plants up to species level. Non-targeted metabolite profiling was performed to determine the putative compound of both plants. Results obtained for rbcL, matK, and ITS DNA barcoding analysis showed 100 % similarity of Oldenlandia corymbosa L. Results obtained for matK DNA barcoding analysis showed 99.64 % similarity of Oldenlandia diffusa (Willd.) Roxb. LC-MS/MS analysis revealed a total of 42 and 12 primary and secondary putative compounds detected in O. corymbosa and O. diffusa extracts, respectively. Overall, we currently recommend the use of both morphological and DNA barcoding for plant identification. In the future, we proposed to design a primer to increase primer specificity and further confirm the structure and biological function of potential putative compounds.
Page 51-57
Muhammad Fazril Mohamad Razif, Boon-Hong Kong, Yeaw-Khim Yee, Noorain Zulkapli, Nik Nurmadihah Azma Nik Ampuan, Shin-Yee Fung
FERMENTED Morinda citrifolia LINN JUICE HAS REDUCED ANTIOXIDANT CAPACITY AND DEMONSTRATES CYTOTOXICITY TOWARDS HUMAN BREAST CANCER CELLS
Abstract
Morinda citrifolia Linn (noni) is a plant belonging to the Rubiaceae family. Recently, fermented noni fruit juice has undergone a surge in popularity, driven by its supposed health benefits. The aim of this study was (i) to determine the nutrient composition of fermented noni fruit juice, and (ii) to compare the antioxidant and cytotoxic properties between unfermented (UNJ) and fermented (FNJ) noni fruit juice. The results showed that FNJ is rich in carbohydrates (81.3 g/100 g DW), but low in both protein (5.4 g/100g DW) and fat (4.0 g/100g DW). FNJ also contains high amounts of amino acids (AAs) glutamic acid, aspartic acid and leucine. UNJ exhibited higher free radical scavenging and antioxidant activity than FNJ. UNJ was found to exhibit comparable cytotoxicity across all cells tested. Notably, FNJ demonstrated higher toxicity against normal human breast (184B5) cells (up to 2.6-fold) compared to human breast cancer (MCF7 and MDA-MB-231) cells. To our knowledge, our study is the first to demonstrate that noni fruit juice (both fermented and unfermented) is toxic to normal breast cells. Their toxicity toward normal breast cells and their non-selective targeting may require in-depth in vivo study to reveal their potential therapeutic benefits. Caution is warranted as noni fruit juice products from other sources may possess different toxicological and pharmacological profiles.
Page 58-67
Mohammad Faruq Abd Rachman Isnadi, Sivan Padma Priya and Tock Hing Chua
PATHOLOGICAL FEATURES OF Plasmodium knowlesi MALARIA INFECTION IN HUMANS AND MACAQUES
Abstract
Plasmodium knowlesi is a simian malaria parasite and currently the dominant species in the Malaysian Borneo of Sabah and Sarawak. This parasite is transmitted by Anopheles balabacensis and the macaques of Macaca fascicularis and Macaca nemestrina, and monkeys of Presbytis melalophos are the reservoir hosts. The zoonotic disease, infection by P. knowlesi infection can cause a wide range of immunological responses in human comparable to other human malaria parasites including acute respiratory distress syndrome (ARDS), acute renal failure (ARF) and cerebral malaria (CM), leading to the pathological consequences in humans and macaques alike. Similar to other malaria species especially P. falciparum, pathological features such as sequestration of parasitised red blood cells (pRBC) and mononuclear cells-containing haemozoin, deposition of haemozoin on numerous tissues and petechial and/or focal haemorrhage could be observed in P. knowlesi infection. Diagnosis of P. knowlesi mainly involves microscopic examination on both thick and thin blood smears stained with Giemsa, nevertheless confirmation test by using Polymerase Chain Reaction (PCR) is compulsory. Most P. knowlesi cases are treated with artemisinin-combination therapy (ACT) or chloroquine in uncomplicated cases while artesunate followed by ACT in complicated or severe cases. Without accurate and timely means of diagnosis and treatment, the outcomes of death might happen. The pathological features of P. knowlesi malaria infection in multiple organs are described in this review.
Page 68-76
Shivangi, Manish Kumar Mishra, Sachin Gupta, Konika Razdan, Shashi Sudan and Shelly Sehgal
ADVANCES IN MOLECULAR VIROLOGY AND DIAGNOSTICS OF HEPATITIS C VIRUS
Abstract
The first and foremost step in the management of any infectious disease, viral or bacterial, is identifying the causative agent behind the disease, followed by determining the extent of incursion attained by it, so as to initiate the specific therapy against and control the exaggerated and self-damaging host reaction. The same applies to the Hepatitis C virus (HCV) infection which is still a global health burden. HCV infection is found worldwide with 71 million people affected globally exhibiting a greater risk of developing advanced-stage liver diseases like fibrosis, cirrhosis and hepatic carcinomas. However, this complex disease state is attained if only the infection is considerably prolonged and left untreated or mistreated. Thus, providing a means to intervene and stop its course, in favour of which an early and accurate diagnosis is required. The timely and correct diagnosis also helps to contain the spread of HCV infection. The basic assays prompting the development of diagnostics are the Enzyme-Linked Immunosorbent (ELISA) and Polymerase Chain Reaction (PCR). On the other hand, recent techniques like microarrays and next-generation sequencing have taken the field of diagnostics to a new phase. Here we present a compilation of all the approaches, recent and older, used in laboratory diagnosis of HCV infection. This review shall help the researchers decide the best method for HCV detection per their requirements and availability of resources.
Page 77-87
Waqas Ahmad and Muhammad Naeem
ANALYSIS OF EFFICIENCY OF SEVEN GENOMIC DNA ISOLATION TECHNIQUES THROUGH APPLICATIONS OF DNA BARCODE, PCR AND NANODROP FROM FINS OF Notopterus notopterus (PALLAS, 1769)
Abstract
Notopterus notopterus is a single species of Genus Notopterus. In Pakistan, N. notopterus is an important fish for food and ornamental trade. The unlimited catch and water pollution are major causes of rapid decline in N. notopterus population. However, genetic study is crucial for its conservation. DNA isolation is the first key step of genetic studies. With this objective, we compared the efficiency of seven genomic DNA isolation techniques. DNA was isolated from fins of N. notopterus because fins were used in small quantities and there was no detrimental effect on fish. Isolated DNA concentration and purity were measured with NanoDrop. PCR amplification and barcoding of mitochondrial COI gene were also used to analyse the purity of isolated DNA. Mitochondrial COI was selected because it is universally used genetic marker for genetic studies of species. Among all methods, GeneJET Genomic DNA Purification Kit was found significantly higher in terms of isolated DNA concentration (894 ng.μl-1), yield (178.3 μg.μl-1), purity (1.7 ng.μl-1), successful PCR amplification and barcode sequencing with 612 base pairs of N. notopterus as compared to investigated six traditional DNA isolation methods. GeneJET Genomic DNA Purification Kit was proved the best in terms of cost and labour and the least hazardous for a handler to perform. Present study has also revealed that the traditional DNA isolated methods are the secondary choice for isolation of DNA from fish fins. Moreover, information about the best genomic DNA isolation technique, from this study can be significant for many molecular techniques such as PCR amplification and gene barcode sequencing among others.
Page 88-92
Yow Hui-Yin, Tang Yin-Quan and Lee Jyy-Shiuan
ANTIPROLIFERATIVE EFFECT OF ASPIRIN AND DICLOFENAC ON MDA-MD-231 TRIPLE-NEGATIVE BREAST CANCER CELLS
Abstract
The role of nonsteroidal anti-inflammatory drugs in triple-negative breast cancer remains unclear. This study aimed to investigate the antiproliferative effect of aspirin and diclofenac on MDA-MB-231 cells using MTT assay. Aspirin (1 – 100 mM) inhibited cell proliferation at dose- and time-dependent manner. At 24 and 48 h treatment, 1 – 100 mM of aspirin exhibited significant antiproliferative effect (p < 0.05). Only ≥ 5 mM aspirin demonstrated significant effect at 72 h (p < 0.05). Similar trend was found with diclofenac (0.01 – 1 mM). This study highlights the potential of aspirin and diclofenac in the management of triple-negative breast cancer
Page 93-103
Maulidi Firlandiana, Giyanto, Wilhelmus Terang Arga Sanjaya, Dwi Andreas Santosa
IN SILICO ANALYSIS OF GENE-ENCODED INVERTASE INHIBITOR (Sininh) FROM SEVERAL SUGARCANE VARIETIES
Abstract
Invertase inhibitor gene fragments have been isolated from various cultivated plants including sugarcane. This article will review the structure of the invertase inhibitor gene (sininh) at 650 bp from multiple sugarcane varieties in Indonesia (PS 881, PS 882, PSJT 941, PS 862, BL/Bulu Lawang, KK/Kidang Kencana). Moreover, the primary structure, physicochemical properties, the secondary structure, 3D structure, and subcellular localization of invertase inhibitor protein (Sininh) were predicted utilizing bioinformatics tools. This information is beneficial for future research, especially in controlling sucrose accumulation at the post-translation level in sugarcane plants. The prediction of the Sininh consisting of 124-145 residues has different characteristics. The predicted Sininh sequences from PS 882 and BL varieties have the potency for translocation due to peptide signals. In addition, there are some Cys residues in both varieties associated with the formation of disulfide bonds for protein structure stability. Sustainable, functional conserved domain Plant invertase/pectin methylesterase inhibitors found in the Sininh protein variety BL/Bulu Lawang where they inhibit the activity of pectin methylesterase (PMEs) invertase with a complex formation. N-glycosylation motifs are also found in the protein Sininh varieties PS 881, PSJT 941, PS 862, BL/Bulu Lawang, KK/Kidang Kencana, which is related to potential stability and interaction with other proteins. These studies build the foundation for studying the structural aspects and the mechanism of the inactivation of invertase via its inhibitory proteins at the molecular level.
Page 104-113
Siew Wei Lee, Joko Logis, Ayesha Fatima, Renee Lay Hong Lim, Szu Ting Ng, Chon Seng Tan, Shin Yee Fung, Nget Hong Tan, Amos Eng Liang Goh, Yong Hui Tan, Crystale Siew Ying Lim
DYE DECOLORISATION CAPACITY OF Lignosus rhinocerotis (Cooke) Ryvarden DIALYSED FRACTION AND ITS EXPRESSION OF NOVEL RECOMBINANT LACCASE IN Pichia pastoris
Abstract
Azo dyes are widely used in the textile industry due to their bold colors and resistance to degradation. This has made azo dyes the global burden in wastewater treatment as conventional dye removal methods have their limitations. Laccase has emerged as an alternative due to its ability to decolorise a wide range of dyes, producing water as a by-product. In this study, we aim to assess the dye decolorisation capacity of the Basidiomycota, L. rhinocerotis dialysed fraction and produce recombinant laccase expressed in Pichia pastoris. Out of the three dyes tested, the dialysed fraction of L. rhinocerotis sclerotia was capable of decolorising around 90% of the Congo Red and Coomassie Brilliant Blue dye at the enzyme unit of 0.018 U/mL. The gDNA of Lac1 [MG210944] was successfully updated in GenBank and the sequence was used to design the pPICZαA/Lac vector, which was successfully cloned and the recombinant Lac1 was successfully expressed using the Pichia pastoris system. The recombinant Lac1 expressed contains the 6x His-tag for purification but the binding towards the Ni-NTA resin was weak, therefore the purification can be optimised through reducing Imidazole concentration to compensate for the weak binding. In conclusion, L. rhinocerotis extract showed promising results in decolorising CR and CBB, while recombinant Lac1 laccase successfully demonstrated its activity and can be further used for dye decolorisation in future studies.