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)
ANNOUNCEMENT
With effect from 15th January 2023, all new submissions will be subjected to a MYR250 publication fee for every accepted paper.
April 2021
Malay. J. Biochem. Mol. Biol. (2021) 24 (1)
TABLE OF CONTENTS
Page 1-6
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Aminu Argungu Umar, Isah Musa Faka, Ibrahim Muhammad Magami
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IN VITRO DNA RESECTION ACTIVITY OF HUMAN EXONUCLEASE1 IS MODULATED BY PCNA AND 14-3-3S
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Abstract
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Cellular response to DNA lesions such as double-strand break (DSB) is dictated by resection of DNA ends. DNA resection is essential for maintaining the genome; without adequate resection of the DNA ends, repair mechanisms such as HR and ATR-dependent checkpoint will not be activated. On the other hand, excessive resection produces unstable single-stranded DNA, which could cause instability of the genome. The regulatory mechanisms of DNA resection are not fully understood. This study was carried out to understand the modulatory effects of PCNA and 14-3-3 proteins on the resection activity of hExo1. Recombinant proteins were expressed from bacteria and purified to near homogeneity. The resection assays were carried out using linearized dsDNA as a substrate. Results obtained from this study show that 14-3-3 proteins bind to hExo1 and inhibit its resection activity, while PCNA interacts with hExo1 and enhances its resection activity. This suggests that the resection activity of hExo1 on dsDNA substrate is modulated through possible protein-protein interactions. The results also show that 14-3-3 proteins repress PCNA binding to hExo1. This study presents an important insight into the mechanism through which DNA resection is controlled. This coordinated control is essential in ensuring efficient and appropriate DNA end resection needed to repair DNA damage to preserve the integrity of the genome.
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Page 7 - 15
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Arief Hidayatullah, Wira Eka Putra, Sustiprijatno, Galuh Wening Permatasari, Wa Ode Salma, Diana Widiastuti, Hendra Susanto, Dewi Ratih Tirto Sari, Febby Nurdiya Ningsih, Muhammad Fikri Heikal, Alyana Mahdavikia Rosyada Yusuf, Aliyya Suci Arizona
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PREDICTING THE POSSIBLE BIOLOGICAL MARKERS AS TARGETED THERAPY FOR DENGUE VIRAL INFECTIONS
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Abstract
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The incidence rate of dengue infections in Indonesia shows an upward trend every year. Recently, dengue infection treatment is still limited to symptom management and especially efforts to restore the body's immune system to fight the infection. There is still no immediate treatment and prevention mechanism for dengue infection. This study aimed to predict the possible targeted proteins in Dengue infection as biomarkers in further molecular assays in part of future drug development. In silico pathway analysis based on KEGG's database was performed to determine the target protein and visualized them into the 3D model through SWISS-Model and PyMOL. Each protein also tested for their interaction using STITCH based on two bioprocesses, namely the cytoplasmic pattern recognition receptor signaling and the innate immune response pathway. The analysis showed that three of the target proteins are RIG-I, MDA5, and MAVS, convincingly interacting in both pathways with very high predictions for specific actions value (0.800), compelling experimental/ biochemical data (0.990 to 0.999), and a low false discovery rate value. RIG-I and MDA5 play an essential role as the viral RNA primary receptors in the early stages of DENV infection. Even by inhibiting these two proteins, it has a severe effect on the cytokine response. The MAVS protein was an essential antiviral protein that serves as a bridge between sensory proteins and responder proteins in innate immune cell mechanisms against DENV infections.
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Page 16 - 33
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Asmaa A. M. Rady, Mohamed A. Abdel Hamid, Heba A. Ewida, Soha M. Hamdy, Marwa G. A. Hegazy, Shadia A. Fathy, Elham A. Hassan, Amany A. Mostafa
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SERUM BIOCHEMICAL CHANGES OF BONE REMODELING FOLLOWING RESIDRONATE- BIOGLASS SCAFFOLDS FOR THE RECONSTRUCTION OF EXPERIMENTALLY INDUCED MANDIBULAR DEFECTS IN DOG MODEL
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Abstract
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Background: Bone tissue engineering has gained significant attention to restoring bone defects by using biodegradable scaffolds. Purpose: This study aimed to evaluate serum biochemical changes of bone remodeling following the use of hybrid scaffolds to reconstruct experimentally induced critical-sized mandibular defects in dogs. Methods: Hybrid scaffolds of chitosan:polyvinyl alcohol named, (CH) alone (no BG) and with bio-glass (BG) graft combined without/with anti-osteoporotic drug (Risedronate M) to form non-medicated and medicated (NM and M) forms were used. A critical size mandibular defect was created into the right mandible of twenty skeletally mature mongrel dogs. These dogs were divided into four equal groups: the 1st group; control (CH) (NM), the 2nd group; CH(M), the 3rd group; CH.BG (NM) and the last group, CH.BG (M). Clinical, radiographic, serum biochemical evaluation (alkaline phosphatase, calcium, phosphorus, calcitonin and osteocalcin) was evaluated at week interval till 12 weeks. Results: A non-significant decrease was found in the activity of osteocalcin in serum for (CH.BG) (M) group when compared to control group CH (NM). There is a highly noteworthy increase in serum calcium, alkaline phosphatase, calcitonin, and phosphorus levels in the (CH.BG) (M) group when compared to the control group CH (NM). Conclusion: Serum biochemical markers of bone healing provided a clinically useful tool to monitor early changes associated with bone healing before radiographic signs of bone healing become obvious. The addition of BG with risedronate drug M improved the degree of bone healing as presented.
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Page 34 - 37
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Catrenar De Silva, Aida Atiqah Mohd Noor, Murni Marlina Abd Karim, Norazah Mohammad Nawawi, Siti Aqlima Ahmad
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PRELIMINARY ANTIBACTERIAL TESTING OF BIOSYNTHESISED SILVER NANOPARTICLES AGAINST THE MARINE AQUATIC PATHOGENS Vibrio alginolyticus AND Vibrio harveyi
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Abstract
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Aquaculture is an up and rising industry that that contributes to the economy of Malaysia. However, fish and shellfish-related diseases have become a leading cause of loss of product and, in turn, loss of money to fish farmers as well as the government. The pathogens that cause these diseases are resistant to commercially available preventive and curing techniques, thus giving rise to new techniques. Biologically synthesised silver nanoparticles are predominantly known for their antibacterial potential and therefore were chosen as the antibacterial agents to combat these pathogens. Forty samples of biosynthesised silver nanoparticles were tested against the marine pathogens Vibrio alginolyticus and Vibrio harveyi for inhibition zones. Samples of Serratia sp. strains AQ5-NT14 and AQ5-NT39 showed the biggest zones and were the most effective against both Vibrio strains.
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Page 38 - 51
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Fatima Gogo Mayaki, Arpah Abu, Abu Bakar Salleh, Siti Nurbaya Oslan
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LipExPS: A VALIDATED WEB-BASED PREDICTION TOOL FOR RECOMBINANT LIPASE EXPRESSION IN Pichia pastoris
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Abstract
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The production of recombinant lipase in Pichia pastoris expression system has become popular due to its reliability and reproducibility. But, choosing the best P. pastoris strain and promoter has been a major issue, leading to time consumption. Hence, there is a need to develop an in silico tool that can assist in the prediction of recombinant lipase production before cloning. LipExPS is a web-based resource that uses a newly curated database (LipExDB) and is devised to provide a platform for predicting the best promoter/host combination to express a lipase gene in P. pastoris. LipExDB was constructed to store lipase expression data that have been successfully expressed in P. pastoris. LipExPS used a BLAST heuristic algorithm to retrieve data from LipExDB to predict the best promoter/host combination when a new lipase is intended to be expressed in P. pastoris system. The competency of LipExPS was experimentally validated using a bacterial lipase gene (ARM lipase). Initially, the amino acid sequence of ARM lipase was submitted into LipExPS followed by evaluating the scores given by the system. Then, based on the given scores and suggestions, the ARM lipase gene was cloned into P. pastoris strain GS115 using alcohol oxidase (AOX) and glyceraldehyde-3-phosphate (GAP) promoters. From the experiment carried out, ARM lipase was successfully expressed in Pichia system using the constructs. Expression of the recombinant ARM lipase in P. pastoris strain GS115 confirmed that LipExPS is a reliable web-based tool to predict the expression of recombinant lipases in P. pastoris.
URL: LipExPS - http://103.18.1.10/lipex/index.html; LipExDB - http://kbioimage.my/lipex/indexDB.php
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Page 52 - 64
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Gregary Chan Chin Khiong, Tansh Rave, Yan See Wan, Adeline Chia Yoke Yin, Yap Wei Hsum, Tang Yin Quan
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BIOLOGICAL PROPERTIES OF STIGMASTEROL AND OTHER PHYTOSTEROLS
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Abstract
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Natural products have long been recognized for their use in traditional medications in mankind. Some of the chemical compounds derived from plants which exhibit high therapeutic values include alkaloids, terpenoids, flavonoid, and phytosterol. Among phytosterols that have been studied, stigmasterol has shown to exhibit significant pharmacological therapeutic values. In this review, the overview of various derivatives' biological properties in the class of phytosterol compounds, especially stigmasterol, will be discussed. A systematic search was employed using google scholar (1977-present) and PubMed (1977- present) based on keywords such as "stigmasterol", "cancer", "structural", "mechanism", "phytosterols", "anticancer", "antimicrobial", "antioxidant", "cardioprotective", "anti- osteoarthritis", "neuroprotective", "anti-inflammatory", and "antidiabetic" to retrieve all relevant research papers published on different biological activities by stigmasterol and its derivatives. This group of compounds exhibits multiple biological effects. Stigmasterol and other identified phytosterol compounds showed promising therapeutic value and high potential candidate drug for various pathological disorders.
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Page 65 - 76
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Ikewuchi, Catherine C., Ifeanacho, Mercy O., Ikewuchi, Jude C
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MODERATION OF DOXORUBICIN-INDUCED DYSLIPIDAEMIA IN WISTAR RATS BY AQUEOUS EXTRACTS OF Pleurotus tuberregium SCLEROTIA AND Cnidoscolus aconitifolius LEAVES
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Abstract
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This study investigated the ability of aqueous extracts of Pleurotus tuberregium sclerotia and Cnidoscolus aconitifolius leaves to prevent or attenuate dyslipidaemia, and adverse alterations in cardiac cholesterol and triglyceride levels, weight gain and heart weight indices, and atherogenic indices, which was induced by intra-peritioneal administration of doxorubicin (15 mg/kg body weight) to Wistar rats. Metformin was administered daily orally at 250 mg/kg; the extracts were administered daily orally at 50, 75 and 100 mg/kg (for 14 days). In both the ameliorative and protective studies, the heart weight, cardiac cholesterol, triglyceride, and plasma triglyceride, apolipoprotein B, total-, non-HDL-, VLDL- and LDL cholesterol concentrations, as well as the atherogenic coefficient, cardiac risk ratio, atherogenic index of plasma and Castelli's risk index II of the Negative control was significantly (p<0.05) higher than those of all the other groups. However, the plasma HDL cholesterol concentration of the Negative control was significantly (p<0.05) lower than those of all the other groups. Therefore, the administration of the extracts and metformin safeguarded (in the protective study) or restored (in the ameliorative study) the doxorubicin induced dyslipidaemia, as well as the cardiac cholesterol and triglyceride levels. These results suggest that, at least in part, due to hypolipidaemic properties, aqueous extracts of Pleurotus tuberregium sclerotia and Cnidoscolus aconitifolius leaves may potentially reduce the risk of doxorubicin-induced cardiovascular disorders.
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Page 77 - 82
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Intan Mariana Maliki, Nor Hasmaliana Abdul Manas, Siti Aqlima Ahmad, Azham Zulkharnain
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EXPRESSION, PURIFICATION AND CHARACTERIZATION OF EXTRADIOL DIOXYGENASE CarBb INVOLVED IN CARBAZOLE DEGRADATION PATHWAY
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Abstract
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Extradiol dioxygenase is a family of enzymes essential for ring cleavage reactions in aromatic compounds degradation pathways. Extradiol dioxygenases belonging to the majority of aromatic compound degradation pathways are single peptide proteins, while a small subset of in this family was reported to be two subunits complex proteins. The extradiol dioxygenase CarB protein is a protein complex consisting of catalytic subunit CarBb with a smaller subunit CarBa. This enzyme was reported to show no ring cleavage activity without the expression of both peptides. However, to date, there was no specific study to confirm CarBb protein dependency on CarBa protein for its ring cleavage activity. In this study, we cloned, heterologously expressed and purified CarBb in E. coli. CarBb protein showed appreciable ring cleavage activity without expression of CarBa protein. The Km and Vmax values calculated were 163.68 μM and 1.19 μM/min. The effects of pH and temperature suggested that the CarBb protein was significantly unstable, suggesting that the CarBa protein may be responsible for the structural stability of the CarBb protein to function as an effective ring cleavage enzyme.
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Page 92 - 99
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Nor Datiakma M.A., Nurhanan Murni Y., Muhammad Haffiz J. and Norjannah S
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PROTEOME PROFILE OF OVARIAN CANCER CELL LINE A2780 TREATED WITH 9-
METHOXYCANTHIN-6-ONE FROM Eurycoma longifolia
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Abstract
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Proteomics is a useful tool for the delineation of the ovarian cancer cell line proteomes expressed under different conditions, enabling these to be interrelated to biological pathways or other aspects of cellular physiology. Since all drug targets are protein in nature, we would like to investigate the differential in protein expression of ovarian cancer cell line A2780 in reaction against 9-methoxycanthin-6-one compound from Eurycoma longifolia. Compound 9-methoxycanthin-6-one was reported in our previous study as having anti-proliferative effects in ovarian cancer cell line. Therefore, in this paper, we extended our study to observe further the modes of action of this compound on ovarian cancer cell lines. Ovarian cancer is the leading cause of mortality in gynaecological malignancies worldwide. It is one of the highest causes of death among female population in Malaysia. In this study, 2-Dimensional Electrophoresis (2-DE) profiling of non-treated and treated ovarian cancer cell line A2780 with 9- methoxycanthin-6-one compound from E. longifolia were performed to observe the in-vitro changes in protein expressions towards understanding the mechanism of action. Differential analyses were performed and subsequently, 15 proteins were selected for proteome identification. Our finding discovered that protein families that affected after ovarian cell line A2780 was treated with 9-methoxycanthin-6-one compound were enzymes, kinase, phosphatase and translation regulator with various molecular functions are amongst the families of proteins involved as adaptive stress response of ovarian cancer cells against 9-methoxycanthin-6-one compound.
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Page 100 - 107
Norsyifa Harun, Illa Syafiqah Johari, Rima Atria Japarin, Siti Najmi Syuhadaa Bakar, Noorul Hamizah Mat, Zurina Hassan, Haniza Hassan
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NALOXONE-PRECIPITATED MITRAGYNINE WITHDRAWAL DID NOT ASSOCIATE WITH INCREASED ANXIETY-LIKE BEHAVIOUR IN RATS
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Abstract
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Kratom (Mitragyna speciosa Korth) is a plant species currently used as an alternative self-treatment for pain and management of opioid dependence and withdrawal. However, information on the effect of its negative emotional state of withdrawal, particularly of the main psychoactive constituent, mitragynine (MG) is still lacking. The present study was designed to determine whether naloxone-precipitated MG withdrawal is associated with increased anxiety-like behaviour in both open-field test (OFT) and elevated-plus maze (EPM) test. Male Sprague Dawley rats were administered with MG and morphine twice daily for 14 consecutive days to develop substance dependence. The withdrawal was precipitated on day fifteen using naloxone (2 mg/kg, i.p.). This study has exhibited that challenge with opioid antagonist naloxone did not result in reliable expression of anxiety behaviour following chronic MG exposure in OFT and EPM tests. However, several somatic signs of withdrawal were observed in naloxone-precipitated MG withdrawn rats. The findings of this study suggest that rat behaviours during OFT and EPM tests might not be driven solely by anxiety levels following naloxone-precipitated withdrawal but rather by other withdrawal-induced behaviours. The presence of somatic signs in naloxone-precipitated MG withdrawn rats was the piece of evidence to prove that that discontinuation of MG or kratom is associated with physical symptoms of withdrawal. However, further studies are prompted to evaluate MG or kratom withdrawal severity to justify its use as opioid replacement therapy.
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Page 108 - 114
Oluwafemi Adeleke Ojo, Adebola Busola Ojo, Odunayo Anthonia Taiwo, Olarewaju Oluba
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NOVEL CORONAVIRUS (SARS-COV-2) MAIN PROTEASE: MOLECULAR DOCKING OF PUERARIN AS A POTENTIAL INHIBITOR
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Abstract
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SARS-CoV-2 is a single-stranded RNA virus that triggered the global pandemic Coronavirus Disease-2019 (COVID-2019). It has infected about 10,021,401 patients and brought forth mortality rate to about 499,913 among 216 countries as cited by WHO. Drugs including Chloroquine and Hydroxychloroquine derivatives are being administered in most urgent cases, although probable side effects on people with metabolic disorders. Thus, the unavailability of authorized drugs and treatment for this pandemic demands the research world to discover natural compounds with potency to cure it. This paper assesses the isoflavonoid puerarin from Pueraria lobata as a possible inhibitor of the main protease of SARS-COV-2 (Mpro) via in silico approach, for example, molecular docking, Lipinski's rule of five and toxicity prediction (ADME). Puerarin revealed a high binding affinity with the target site of SARS-CoV-2 main protease. This compound slightly meets the criteria of Lipinski's rule. It does not possess properties that could cause adverse effects in humans; thus, making puerarin a potential drug candidate to investigate its usage against COVID-19.
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Page 115 - 128
Pavan Gollapalli, Aditya Rao SJ, Shivananda Kandagalla, Sharath Belenahalli Shekarappa, Manjunath Chavadi, Manjunatha Hanumanthappa
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INSIGHTS INTO THE DYNAMIC DOMAIN FLUCTUATIONS AND SIGNAL PROPAGATION OF ALLOSTERIC REGULATION IN PYRUVATE KINASE IN Mycobacterium tuberculosis
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Abstract
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The enzyme's structure-based function relationship suggested that pyruvate kinase regulation acts as a pivotal step in the energy management of Mycobacterium tuberculosis during infection. The present study aims to understand the network of interactions between the ligands Adenosine monophosphate (AMP) and Glucose-6-phosphate (G6P) during their allosteric regulation of pyruvate kinase (pyk) from M. tuberculosis with its imposed structural stability via dynamic domain fluctuations and signal propagation using molecular dynamics simulations studies. The results revealed that the molecular re-arrangement and associated conformational modifications after the smooth binding of AMP and G6P to pyruvate kinase result in reduced enzyme flexibility. Further, based on the signal propagation rate, the residues involved in the mediation of allosteric communication between the domains during fluctuation to regulate pyruvate kinase enzyme allosterism were predicted. The synergistic assistance of two allosteric binding sites for AMP and G6P in M. tuberculosis pyk monomer provides a piece of evidence for the existence of two physical allosteric pathways in the bacterium to regulate several metabolic processes.
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Page 129 - 137
Thiban Sandramuti, Nur Fatin Zalikha Zailan, Masriana Hassan
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ATTENUATION OF CD11B EXPRESSION AND PRO-INFLAMMATORY RESPONSES VIA GLYCOGEN SYNTHASE KINASE-3 INHIBITION IN LIPOPOLYSACCHARIDE STIMULATED BV2 MICROGLIAL CELLS
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Abstract
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Excessive production of pro-inflammatory mediators by microglia may lead to neuroinflammation and neuronal death. Glycogen synthase kinase-3 (GSK-3) is a regulatory enzyme that involves in the mechanism of pro- and anti-inflammatory responses. Inhibition of GSK-3 potentially attenuates the aggravation of neuroinflammation. Objective: We aimed to investigate the effects of GSK-3 inhibition on the immune responses through assessment of the expressions of CD11b and production of immune mediators in lipopolysaccharide (LPS)-stimulated BV2 microglial activation. The activation status of BV2 stimulated with LPS was determined by the expression of CD11b through flow cytometry analysis. The production and expression of nitric oxide (NO) and pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), and monocyte chemoattractant protein 1 (MCP-1) in the absence or presence of GSK-3 inhibitors namely Tideglusib (TG) and lithium chloride (LiCl) in LPS-stimulated BV2 cells were determined by using Griess assay, BD cytometric beads array mouse inflammatory kit, and quantitative RT-PCR (qPCR), respectively. Activation of microglia through stimulation with LPS significantly increased the expression of CD11b and the production and expression of NO and pro-inflammatory cytokines. The expression of CD11b reduced approximately 17% and 23% (p<0.001) when treated with TG and LiCl, respectively. Analysis of NO production in response to GSK-3 inhibitor showed a significantly decreased by approximately 50% (p<0.001) indicated by changes in nitrite levels. Additionally, inhibition of GSK-3 significantly decreased the production of TNF-α, IL-6, and MCP-1 as observed in the presence of 10 mM and 15 mM of LiCl (p<0.05) or 5 μM and 10 μM of TG (p<0.001). The reduction of these pro-inflammatory cytokine levels is corroborated by a significant decline in inducible nitric oxide synthase (iNOS), CD11b, TNF-α, and IL-6 gene expression. Inhibition of GSK-3 signaling molecule potentially downregulate the neuroinflammation through attenuation of the production and expression of NO, pro-inflammatory cytokines, and CD11b elicited by LPS in microglia.
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Page 138 - 142
Vishakh R, Suchetha Kumari, Tamizh Selvan G
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ASSOCIATION OF SERUM VITAMIN D LEVELS IN EARLY AND LATE DIABETES – A REVIEW
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Abstract
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Diabetes mellitus is one of the common chronic conditions, which affects millions of people around the world. It is associated with many problems, including micro and macrovascular abnormalities, which not only impair the quality of life significantly but also increase mortality. There has been widespread interest in the pathogenesis and prevention of vitamin D-related diabetes. Vitamin D deficiency has become increasingly recognized all around the world in recent years. Above and beyond its classical role as a regulator of calcium and phosphorous metabolism and bone mineralization, vitamin D facilitates the secretion of insulin from the beta cells of the pancreas to increase glucose uptake by the peripheries and decrease systemic inflammation. Thus, vitamin D deficiency has been found to have a vital role in the pathogenesis of diabetes mellitus. Vitamin D deficiency is known to be associated with glucose intolerance, insulinemia, insulin resistance, and obesity. The majority of patients with diabetes mellitus may have low serum vitamin D levels. Vitamin D supplementation has been shown to reduce insulin resistance in diabetes mellitus. The normal serum level of vitamin D and the range of related factors are still essential to carry out an extensive investigation to conclude a risk factor. In anticipation of that, it is probable to conclude that maintenance of serum vitamin D levels holds benefits in diabetic disorders, and researchers have to be duty-bound to attempt to attain them.
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Page 143 - 147
Wira Eka Putra, Sustiprijatno, Wa Ode Salma, Diana Widiastuti
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DISRUPTING THE INTERACTION OF IL-1/IL-1 RECEPTOR WITH Morinda officinalis BIOACTIVE COMPOUNDS AS STRATEGY TO COMBAT CHRONIC INFLAMMATORY DISEASES: THE INSIGHT FROM IN SILICO STUDY
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Abstract
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Chronic inflammatory diseases are serious health problems worldwide. Furthermore, this entity leads to several types of diseases, including cancer, cardiovascular diseases, autoimmune diseases, and diabetes. Many modalities have been proposed to ameliorate chronic inflammatory diseases, including the use of traditional medicine. Recently, the use of herbs as traditional medicine becomes popular in all aspects of society. One of the most plants that are widely used is Morinda officinalis. Importantly, M. officinalis exerts its medical properties in multiple biological activities, including anti-cancer, anti-inflammatory, and even cardiovascular activity. Thus, in this study, we aimed to evaluate the bioactive compounds of M. officinalis through in silico approach as an inhibitor of interleukin-1/ interleukin-1 receptor (IL-1/IL-1R) interaction to ameliorate the chronic inflammatory diseases. Molecular docking was used as prediction methods of interaction between the ligands and target protein. In this present study, we found three compounds, namely 1,2-dihydroxy-3-methylanthraquinone, rubiadin, and rubiadin-1-methyl ether, the greatest score of binding affinity (-7.6 kcal/mol) to the target protein. Data visualization also showed a similar interaction pattern, which ensures they have similar binding areas in the target protein, the IL-1R. Therefore, this prediction suggested that these three compounds might provide potential as an inhibitor against the IL-1R. Further research needs to be done to clarify and explore more details about the biological activity of these compounds, especially in chronic inflammatory diseases.
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Page 148 - 159
Nur Aina Shazwani Sukri, Zamzahaila Mohd Zin, Mohd Aidil Adhha Abdullah, Nor Dini Rusli, Katherine Louise Smedley, Mohamad.Khairi Zainol
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EFFECT OF DIFFERENT PRE-SOAKING TECHNIQUE ON PHYSICOCHEMICAL PROPERTIES OF Canavalia ensiformis PROTEIN HYDROLYSATE
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Abstract
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Canavalia ensiformis or Kacang Koro (Jack Beans) is one of the under-exploited tropical beans that contains between 25-35% crude protein content. Pre-treatments such as soaking helps to maintain the desired properties of protein hydrolysate, such as antioxidants, structure, high water capacity, oil-water capacity, and removing antinutrients. However, there is little information on the effect of soaking using different solvents on physicochemical properties of C. ensiformis. The main objectives of this study are to determine the effect of pre-treatments (A= lime juice, B=in 50% w/v tamarind solution (1:2) for 15 min and rinsed in distilled water, C=3% salt solution (1:3) and 50% (w/v) activated carbon solution (1:2), D= in distilled water (1:2) as control.) on physicochemical properties of C. ensiformis protein hydrolysate. The hydrolysates were evaluated based on moisture content, protein content, water holding capacity (WHC), oil holding capacity (OHC), emulsifying properties, solubility, degree of hydrolysis (DH), foam stability, foaming capacity and antioxidant properties using 2,2-diphenyl-2-picrylhydrazyl (DPPH) assay for antioxidant activities. The highest protein content (6.89±0.71%) was observed in treatment D. Treatment C exhibited the highest degree of hydrolysis water, oil holding capacity, emulsifying stability index (ESI), solubility and DPPH. The size of C. ensiformis protein hydrolysate analysed using scanning electron microscope (SEM) varies due to different pH soaking treatments between pH 5 to pH 10. C. ensiformis protein hydrolysate treated in treatment B showed the highest inhibition towards DPPH (27.23±4.65%). Overall, this study showed that the best technique for extracting a high quality C. ensiformis protein hydrolysate that can be used in food or pharmaceutical products was pre-soaking in salt and activated carbon, thus promoting the use of this under-utilized legume.
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Page 160 - 168
Subashini Muniandy, Muhammad Amin Sanusi, and Suriani Mohamad
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THERMOSTABLE ACYL HOMOSERINE LACTONASE FROM Bacillus cereus SM01 AND ITS QUORUM-QUENCHING ACTIVITIES ON Burkholderia pseudomallei AND METHICILLIN RESISTANT Staphylococcus aureus
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Abstract
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N-acyl homoserine lactone (AHL) plays an important role in bacterial quorum sensing. Bacterial cells use AHL molecules to coordinate the expression of various virulence determinants and biofilm development within the population. Therefore, quenching the activity of these molecules offers an alternative strategy to control bacterial infection. In this study, isolate Bacillus cereus SM01 was found to possess enzymes that could inactivate AHL molecules, rendering them unfit for quorum sensing. This study describes the characterization and quorum quenching activities of recombinant AHL-lactonase (AiiA) cloned from B. cereus SM01. Using gene-specific primers, a 753 base pair AiiA homologue that codes for AHL-lactonase were successfully amplified from B. cereus SM01 genomic DNA and cloned into the pCOLD expression system. Homology comparison of the deduced amino acid of the cloned DNA sequence showed that it was highly identical with known AHL-lactonases from the Bacillus group and belonged to the metallo-beta-lactamase superfamily of proteins. The resultant 29kD recombinant AHL lactonase protein possessed broad pH tolerance (from pH 5 up to 11) and good thermal stability. Prolonged exposure of the recombinant AHL-lactonase to high temperatures (65oC or 85oC, 1 hour) did not diminish the AHL-inactivating activity. It successfully quenched the activity of the quorum-sensing molecules of Burkholderia pseudomallei (B. pseudomallei) UKMS01. The recombinant AHL lactonase also significantly reduced biofilm development in methicillin-resistant Staphylococcus aureus and B. pseudomallei by 3.4 folds and 4.8 folds, respectively.
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Page 169 - 174
Tebekeme Okoko, Solomon Ameniyegba Ndoni
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PROTECTIVE EFFECT OF KOLAVIRON ON BROMATE-INDUCED TOXICITY ON RAW U937 CELLS AND MACROPHAGES
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Abstract
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Bromate is used as an additive in bread making and also a disinfection by-product during water purification. However, its reported toxicity seems to limit its usage. The protective effect of kolaviron on bromate-induced toxicity on the cell line U937 and U937-derived macrophages was investigated. In the first instance, cell culture was supplemented with kolaviron before incubated with bromate. Later cell viability and the production of reactive oxygen species (ROS) were investigated using the MTT reduction and DCHF-DA assays, respectively. In the other experiment, the U937 cells were first differentiated to the macrophage form using phorbol-12-myristate-13-acetate. After that, culture having the U937-derived macrophages was supplemented with kolaviron before exposure to bromate. Later, the production of TNF-α and IL-6 was assessed via cytokine ELISA. The expression of the transcription factors iNOS and NF-κB was also determined via RT-PCR. It revealed that kolaviron protected the cells from bromate-induced cytotoxicity and ROS production significantly compared to controls (p < 0.05). Pre-incubated the U937-derived macrophages with kolaviron also reduced the bromate-mediated production of TNF-α and IL-6. Kolaviron also reduced bromate-induced expression of iNOS and NF-κB in the macrophages. The kolaviron-mediated response on cytotoxicity, production of ROS and TNF-α was concentration-dependent (p < 0.05). The findings show that the reduction by kolaviron on bromate-induced cytotoxicity and production of reactive oxygen species could be linked to the modulation of the inflammatory response.
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Page 175 - 184
Zanudin NAM, Hasan NA, Noruddin NFN, Wakid SA, Hasbullah NI
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PRELIMINARY SCREENING OF ENDOPHYTIC FUNGI FROM Capsicum annuum L. FOR BIOCONTROL ACTIVITY AGAINST Colletotrichum gloeosporoioides
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Abstract
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Anthracnose is a plant disease caused by the Colletotrichum sp., which responsible for declining the production of chilli production worldwide. Farmers have been using various agronomic practices in managing plant diseases, such as planting resistant crop varieties, applying chemical fungicides and introducing biological control agents. However, due to the drawbacks of chemical fungicide usage, biological control has become an alternative option. This study was conducted to determine the potential endophytic fungi isolated from chilli as biocontrol agents against phytopathogen, Colletotrichum gloeosporoioides. In the present work, we isolated, identified and characterized a total of 85 fungal isolates harboring inside the leaves, fruits and stems of Capsicum annuum L. The endophytic fungi isolates were most frequently found in the leaves of C. annuum (1.17) compared to stems (0.24) and fruits (0.08). The cultural-morphological identification was grouping these isolates into 14 groups. Based on the rDNA ITS gene sequence, 14 fungal isolates belonged to Phylum Ascomycota. Most of the dominating fungal endophytes were from the genus Trichoderma. For antagonistic test, Hypoxylon sp. (F2) and Trichoderma reesei (F4) isolates recorded a promising potential against C. gloeosporoioides phytopathogens with 87.21% and 76.26% of radial growth inhibition, respectively. The finding in this study suggested the initial step in utilizing the isolates for further development of these endophytic fungal as antagonists and antibacterial agents for future biotechnological applications in chilli growing fields.
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