Pusat Penyelidikan Biokimia Analisis - Tesis

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Synthesis Of Choline Chloride-acetic Acid Based Deep Eutectic Solvent (Des) For The Extraction Of Bioactive Polysaccharide From Garcinia Atroviridis Fruit
(2024-09)
Razak, Nur Qistina Abdul
The fruit of Garcinia atroviridis was identified as a potential source of bioactive polysaccharides. This study explored the ability of these polysaccharides (referred to as GAP) to inhibit pancreatic lipase and angiotensin-converting enzyme (ACE), both of which are important in managing obesity and hypertension. The extraction of GAP was optimized using a microwave-assisted process involving deep eutectic solvents (DES). The study was promoted to prepare deep eutectic solvents (DES) from choline chloride and acetic acid in 1:2, 1:3, 1:4. The physicochemical properties of the resulting DES were fully characterized. The interaction of the DES components by hydrogen bonds was confirmed using Fourier-transform infrared spectroscopy (FTIR) and changes in wavenumbers of their functional groups. Moreover, these DES were shown to be hydrophilic and could dissolve in water with high polarity while having compatibility with many of semi-polar solvents. the DES demonstrates excellent potential for extraction applications due to its low pH and polar nature. Therefore, DES is a promising medium for extracting GAP. The yield of GAP was then extracted and the result concluded that the experimental yield of GAP (24.33%) closely matched the predicted yield (24.88%), confirming the accuracy of the extraction model. GAP was found to have a high molecular weight (28.15 ± 3.17 kDa) and viscosity (3.171 ± 1.23 Pa·s), along with a significant branching size (6.39 ± 0.030). These characteristics suggest that GAP can form complex structures that effectively inhibit enzyme activity by physically blocking the enzymes from accessing their substrates, thereby slowing down their function. In laboratory tests, GAP demonstrated significant inhibitory effects, reducing pancreatic lipase activity by 79.41% and ACE activity by 81.23%. These results indicate that GAP has strong potential for use in developing anti-obesity and antihypertensive treatments. Further optimization of GAP's bioactivity led to even better results, with optimized GAP inhibiting pancreatic lipase (GAP-L) by 87.93% and GAP that inhibits ACE (GAP-A) by 95.91%. The experimental values closely aligned with the predicted values (86.57% for pancreatic lipase inhibition and 95.64% for ACE inhibition), further validating the model. Therefore, GAP shows great promise as a potential source for developing effective agents to manage obesity and hypertension. The suitability of the DES produced as well shown to have a polar nature, making it capable of interacting with and dissolving a wide range of polar compounds, including polysaccharides and other bioactive molecules.
Microwave-Assisted Extraction Of Polysaccharide From Cinnamomum Cassia With Anti-Hyperpigmentation Properties
(2023-06)
Al Ajalein Alhareth Abdulraheem Salem
Polysaccharides with anti-hyperpigmentation properties have not been reported elsewhere. The current study extracted the polysaccharides from Cinnamomum cassia bark using microwave-assisted approach, and optimized the extraction parameters (i.e. microwave power, irradiation time and buffer-to-sample (B:S) ratio) based on Box-Behnken design to obtain polysaccharides with high anti-hyperpigmentation activities, antioxidant activity, sun protection factor (SPF) as well as the extraction yield.
Elucidation And Comparison Of Proteome Profiles In Normal, Precancerous And Cervical Cancer Tissues
(2024-04)
Gaayathri A/P Kumarasamy
The aim of this study was to determine the protein profiles and differentially expressed proteins in the normal cervix, HPV-associated squamous intraepithelial lesion (SIL) which is precancerous and cervical cancer, extracted from formalin-fixed paraffin-embedded (FFPE) tissues.
Identification Of Bioactive Peptide From Chicken Ovalbumin Using An Integrated Bioinformatics-assisted Approach And Determining Their Functional Significance
(Universiti Sains Malaysia., 2021-10)
Mohd Adam Salim Mohd Salim
complete integrated bioinformatics approach was developed to identify angiotensin-converting enzyme (ACE) and dipeptidyl peptidase-4 (DPP-4) inhibitory peptides from chicken ovalbumin (OVA). This approach involved PeptideCutter, Peptide Ranker and Pepsite2 in order to hydrolyse OVA protein sequence into smaller peptides, to identify the probability of the peptides being bioactive and to investigate the interaction between the peptides and target enzymes (i.e. ACE and DPP-4), respectively. OVA sequence was initially hydrolysed using PeptideCutter. Pepsin (P), Chymotrypsin (C) and Trypsin (T) were used in 7 different OVA (O) hydrolysis combination (OP, OC, OT, OCT, OPC, OPT and OPCT), thus, producing 71 peptides. Top ten novel bioactive peptides (i.e. CF, KM, ELPF, AM, ADHPH, LPR, PR, FR, PRM and GR) were then successfully identified and selected based on the amino acid sequences as well as the peptide interactions with ACE and DPP-4. Against ACE, IC50 of CF, KM, ELPF, AM, ADHPH, LPR, PR, FR, PRM and GR were 1.82, 1.89, 4.24, 3.07, 3.54, 1.30, 5.47, 4.35, 5.22 and 3.11 mM, respectively. These results were comparable to commercial inhibitor for ACE, captopril (IC50 = 3.98 mM). While against DPP-4, however, inhibitory activities were only comparable to other reported DPP-4 inhibitory peptides such as EK (IC50 = 3.22 mM) and GL (IC50 = 2.62 mM), as the peptides were able to achieve 2.99, 2.22, 9.92, 2.79, 1.66, 1.43, 4.11, 2.47, 2.50 and 2.83 mM, respectively
Characterisation And Crystallisation Of Deptor
(Universiti Sains Malaysia, 2021-02)
Yeap, Kean Heng
The mechanistic target of rapamycin (mTOR) signalling pathway integrates wide varieties of nutrients to regulate important cellular processes related to cell growth. mTOR is a huge serine/threonine protein kinase and exists in two distinct complexes, mTORC1 and mTORC2. DEPTOR is an endogenous inhibitor of mTOR and consists of two DEP domains in tandem arrangement and a PDZ domain. The Escherichia coli expression system was used for cloning and expression experiments. Here, besides the full-length protein, various truncated mutants were designed as well to gain insights into the characteristics and architectures of the protein. Four truncated mutants were constructed to represent the first DEP domain (DEP1), the second DEP domain (DEP2), both DEP domains (DEP12) and the PDZ domain. DEPTOR, DEP1, DEP2 and DEP12 were expressed as soluble proteins but PDZ was insoluble.

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