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Optimization of Pectin Extraction from Sweet Potato Peels Using Citric Acid and its Emulsifying Properties

Hamidon NH, Abang Zaidel DN, Mohd Jusoh YM

Recent Pat Food Nutr Agric, 2020;11(3):202-210.
PMID: 32031081 DOI: 10.2174/2212798411666200207102051

BACKGROUND: Pectin is a natural polysaccharide that has been used widely as a stabilizer in food emulsion system.
OBJECTIVE: This study aimed to optimize the yield of pectin extracted from sweet potato residue and investigate its emulsifying properties.
METHODS: Response surface methodology (RSM) has been utilized to investigate the pectin extracted from sweet potato peels using citric acid as the extracting solvent. Investigation of the effect of different extraction conditions namely temperature (°C), time (min) and solution pH on pectin yield (%) were conducted. A Box-Benhken design with three levels of variation was used to optimize the extraction conditions.
RESULTS: The optimal conditions determined were temperature 76°C, time 64 min and pH 1.2 with 65.2% yield of pectin. The degree of esterification (DE) of the sweet potato pectin was determined using Fourier Transform Infrared (FTIR) Spectroscopy. The pectin is high-methoxyl pectin with DE of 58.5%. Emulsifying properties of sweet potato pectin were investigated by measuring the zeta-potential, particle size and creaming index with addition of 0.4 and 1.0 wt % pectin to the emulsion.
CONCLUSION: Extraction using citric acid could improve the pectin yield. Improved emulsion stability was observed with the addition of the sweet potato pectin.
Generation and selection of naïve Fab library for parasitic antigen: Anti-BmSXP antibodies for lymphatic filariasis

Omar N, Hamidon NH, Yunus MH, Noordin R, Choong YS, Lim TS

Biotechnol Appl Biochem, 2018 May;65(3):346-354.
PMID: 28833498 DOI: 10.1002/bab.1591

Phage display has been applied successfully as a tool for the generation of monoclonal antibodies (mAbs). Naive antibody libraries are unique as they are able to overcome several limitations associated with conventional mAb generation methods like the hybridoma technology. Here, we performed an in vitro selection and generation of Fab antibodies against Brugia malayi SXP protein (BmSXP), a recombinant antigen for the detection of lymphatic filariasis. We developed a naïve multi ethnic Fab antibody library with an estimated diversity of 2.99 × 109 . The antibody library was used to screen for mAbs against BmSXP recombinant antigen. Soluble monoclonal Fab antibodies against BmSXP were successfully isolated from the naïve library. The Fab antibodies obtained were expressed and analyzed to show its binding capability. The diversity obtained from a pool of donors from various ethnic groups allowed for a diverse antibody library to be generated. The mAbs obtained were also functional in soluble form, which makes it useful for further downstream applications. We believe that the Fab mAbs are valuable for further studies and could also contribute to improvements in the diagnosis of filariasis.
Immune TB Antibody Phage Display Library as a Tool To Study B Cell Immunity in TB Infections

Hamidon NH, Suraiya S, Sarmiento ME, Acosta A, Norazmi MN, Lim TS

Appl Biochem Biotechnol, 2018 Mar;184(3):852-868.
PMID: 28884285 DOI: 10.1007/s12010-017-2582-5

B cells and in particular antibodies has always played second fiddle to cellular immunity in regard to tuberculosis (TB). However, recent studies has helped position humoral immunity especially antibodies back into the foray in relation to TB immunity. Therefore, the ability to correlate the natural antibody responses of infected individuals toward TB antigens would help strengthen this concept. Phage display is an intriguing approach that can be utilized to study antibody-mediated responses against a particular infection via harvesting the B cell repertoire from infected individuals. The development of disease-specific antibody libraries or immune libraries is useful to better understand antibody-mediated immune responses against specific disease antigens. This study describes the generation of an immune single-chain variable fragment (scFv) library derived from TB-infected individuals. The immune library with an estimated diversity of 109 independent clones was then applied for the identification of monoclonal antibodies against Mycobacterium tuberculosis α-crystalline as a model antigen. Biopanning of the library isolated three monoclonal antibodies with unique gene usage. This strengthens the role of antibodies in TB immunity in addition to the role played by cellular immunity. The developed library can be applied against other TB antigens and aid antibody-derived TB immunity studies in the future.
Fulltext Microencapsulation of Lactococcus lactis Gh1 with Gum Arabic and Synsepalum dulcificum via Spray Drying for Potential Inclusion in Functional Yogurt

Fazilah NF, Hamidon NH, Ariff AB, Khayat ME, Wasoh H, Halim M

Molecules, 2019 Apr 11;24(7).
PMID: 30978923 DOI: 10.3390/molecules24071422

There has been an explosion of probiotic incorporated based product. However, many reports indicated that most of the probiotics have failed to survive in high quantity, which has limited their effectiveness in most functional foods. Thus, to overcome this problem, microencapsulation is considered to be a promising process. In this study, Lactococcus lactis Gh1 was encapsulated via spray-drying with gum Arabic together with Synsepalum dulcificum or commonly known as miracle fruit. It was observed that after spray-drying, high viability (~10⁸ CFU/mL) powders containing L. lactis in combination with S. dulcificum were developed, which was then formulated into yogurt. The tolerance of encapsulated bacterial cells in simulated gastric juice at pH 1.5 was tested in an in-vitro model and the result showed that after 2 h, cell viability remained high at 1.11 × 10⁶ CFU/mL. Incubation of encapsulated cells in the presence of 0.6% (w/v) bile salts showed it was able to survive (~10⁴ CFU/mL) after 2 h. Microencapsulated L. lactis retained a higher viability, at ~10⁷ CFU/mL, when incorporated into yogurt compared to non-microencapsulated cells ~10⁵ CFU/mL. The fortification of microencapsulated and non-microencapsulated L. lactis in yogurts influenced the viable cell counts of yogurt starter cultures, Lactobacillus delbrueckii subs. bulgaricus and Streptococcus thermophilus.
Bioassay-Guided Fractionation of Acetone and Methanol Extracts of Quercus infectoria Galls with Antimalarial Properties

Hamidon NH, Dona ACT, Zin NNINM, Nordin NI, Sulaiman SF, Abu-Bakar N

Trop Life Sci Res, 2024 Jul;35(2):167-185.
PMID: 39234468 DOI: 10.21315/tlsr2024.35.2.8

The antimalarial properties of crude extracts from Quercus infectoria galls were investigated through bioassay-guided fractionation. Acetone (QIA) and methanol (QIM) crude extracts have been reported to have promising antimalarial activity against Plasmodium falciparum (3D7 strain). These extracts were subjected to fractionation using automated preparative high-performance liquid chromatography (prep-HPLC) to identify the most active fractions. Nine fractions were isolated from each extract, of which the fractions QIA11 and QIM16 showed antimalarial activity, with IC50 values of 17.65 ± 1.82 μg/mL and 24.21 ± 1.88 μg/mL, respectively. In comparison, the standard antimalarial drug artemisinin has an IC50 value of 0.004 ± 0.001 μg/mL). Through high-resolution liquid chromatography coupled with mass spectrometry (HR-LCMS) analysis of the fractions, four known compounds were successfully identified: gallic acid, ellagic acid, 1,3,6-tris-o-(3,4,5-trihydroxybenzoyl)-beta-d-glucose and 1-O,6-O-digalloyl-beta-D-glucose.