alpha-Galactosidase (EC 3.2.1.22) from ripe papaya (Carica papaya L.) fruit was fractionated by a combination of ion exchange and gel filtration chromatography into three forms, viz., alpha-galactosidase 1, 2 and 3. The predominant isoform, alpha-gal 2, was probably a tetramer with a native molecular mass of about 170 kDa and 52 kDa-sized subunits and an estimated pI of 7.3. The subunit's N-terminal amino acid sequence shared high identity (97%) with the deduced sequence of a papaya cDNA clone encoding a putative alpha-galactosidase PAG2 as well as with an Ajuga reptans L. GGT1 clone encoding a galactan: galactan galactosyltransferase (66%). During ripening, alpha-galactosidase activity increased concomitantly with firmness loss and this increase was largely ascribed to alpha-gal 2. The protein level of alpha-gal 2 as estimated by immunoblot was low in developing fruits and generally increased with ripening. alpha-Galactosidase 2 also had the ability to markedly catalyse increased pectin solubility and depolymerisation while the polymers were still structurally attached to the cell walls mimicking, in part, the changes that occur during ripening. The close correlation between texture changes, alpha-gal 2 activity and protein levels as well as capability to modify intact cell walls suggest that the enzyme might contribute to papaya fruit softening during ripening. The purported mechanism of alpha-gal 2 action as a softening enzyme was discussed in terms of its functional capacity as a glycanase or perhaps, as a transglycosylase.
Volatile compounds in the peel of calamansi (Citrus microcarpa) from Malaysia, the Philippines and Vietnam were extracted with dichloromethane and hexane, and then analysed by gas chromatography-mass spectroscopy/flame ionisation detector. Seventy-nine compounds representing >98% of the volatiles were identified. Across the three geographical sources, a relatively small proportion of potent oxygenated compounds was significantly different, exemplified by the highest amount of methyl N-methylanthranilate in Malaysian calamansi peel. Principal component analysis and canonical discriminant analysis were applied to interpret the complex volatile compounds in the calamansi peel extracts, and to verify the discrimination among the different origins. In addition, four common hydroxycinnamic acids (caffeic, p-coumaric, ferulic and sinapic acids) were determined in the methanolic extracts of calamansi peel using ultra-fast liquid chromatography coupled to photodiode array detector. The Philippines calamansi peel contained the highest amount of total phenolic acids. In addition, p-Coumaric acid was the dominant free phenolic acids, whereas ferulic acid was the main bound phenolic acid.
Calamansi juices from three countries (Malaysia, the Philippines and Vietnam) were characterised through measuring volatiles, physicochemical properties and non-volatiles (sugars, organic acids and phenolic acids). The volatile components of manually squeezed calamansi juices were extracted using dichloromethane and headspace solid-phase microextraction, and then analysed using gas chromatography-mass spectrometry/flame ionisation detector, respectively. A total of 60 volatile compounds were identified. The results indicated that the Vietnam calamansi juice contained the highest amount of volatiles. Two principal components obtained from principal component analysis (PCA) represented 89.65% of the cumulative total variations of the volatiles. Among the non-volatile components, these three calamansi juices could be, to some extent, differentiated according to fructose and glucose concentrations. Hence, this study of calamansi juices could lead to a better understanding of calamansi fruits.
Microcrystalline cellulose (MCC) extracted from empty fruit bunches (EFB), stalk and spikelet were characterised through physicochemical and microstructure analyses. Raw stalk fibres yielded the highest cellulose content (42.43%), followed by EFB (32.33%) and spikelet (18.83%). Likewise, lowest lignin and residual oil content was reported in raw stalk fibres compared to EFB and spikelet. SEM revealed significant changes on fibres' surface morphology throughout the extraction process. FTIR analysis showed that main characteristic peaks of hemicellulose and lignin was absent on the extracted MCC. The crystallinity index for MCC extracted from EFB (82.5%), stalk (82.2%) and spikelet (86.5%) was comparable to commercial MCC (81.9%). Results suggested stalk fibres is more preferable for the production of MCC compared to EFB and spikelet. Further rheological studies showed viscoelastic behaviour with no significant differences between commercial and stalk-based MCC, while modelling work showed ability to simulate complex deformation of the MCC-hydrogel/food mixture during processing/handling stage.
After oil extraction, palm fruit biomass contains abundant water-soluble phytochemicals (PCs) with proven bioactivity in regulating oxidative stress and inflammation (OSI). For optimal bioefficacy following oral consumption, the pharmacokinetic plasma peak (Tmax) should be bio-matched with the onset of OSI, which can be predicted from the Phytochemical Absorption Prediction (PCAP) model and methodology. Predicted absorption and potential for regulation of OSI by measures of total phenolic content, antioxidant capacity and hydrogen peroxide production capacity, were applied to characterise eight extracts from mesocarp fibre and kernel shells of oil-depleted palm fruits. Results indicated post-consumption absorption Tmax ranges of 0.5-12 h and 2-6 h for intake in liquid and solid forms, respectively, and generally high antioxidant activity of the extracts. The research supports that PC extracts of palm fruit biomass have broad potential uses for human health as dietary antioxidants in foods, supplements or functional beverages.
Threadfin bream (Nemipterus japonicas) muscle was hydrolysed using protease extracted from
bilimbi (Averrhoa bilimbi L.) fruit. This study was performed in order to compare the efficiency of bilimbi protease in producing threadfin bream protein hydrolysate with the commercial protease; alcalase 2.4 L. Initially, protease was extracted and then purified using 40% ammonium sulfate precipitation method. The proteolytic activity of the crude extract and purified protease was determined. Precipitation using 40% ammonium sulfate resulted in bilimbi protease specific activity of 2.36 U/mg and 23.13% recovery. Threadfin bream hydrolysate was prepared based on the pH-stat method by hydrolysis for 2 hrs. Hydrolysis using bilimbi protease produced 34.76% degree of hydrolysis (DH) and 3.75% yield while hydrolysis using alcalase resulted in 86.6% DH with 22.78% yield. Alcalase hydrolysate showed higher solubility than bilimbi protease hydrolysate at pH 7 with 70.87 and 32.16% solubility, respectively. Results also showed that protein content of threadfin bream hydrolysate produced using alcalase was higher (86.86%) than those produced using bilimbi protease (22.12%). However, both hydrolysates showed low moisture content between 3.93 to 7.00%. The molecular weight distribution analysis using SDS–PAGE indicated the distribution of smaller peptides especially in alcalase hydrolysate. Overall, the results showed that alcalase is more efficient enzyme choice than bilimbi protease for preparing threadfin bream hydrolysates. However, both hydrolysates could play an important role thus contribute to the food industry.
Oil palm empty fruit bunch graft poly (acrylic acid-co-acrylamide) superabsorbent composite (OPEFB-g-(PAA-co-PAM) SAPC) was synthesized by graft copolymerization of the acrylic acid (AA) and acrylamide (AM) comonomer onto OPEFB fibre using ammonium persulfate (APS) and N,N-methylene bisacarylamide (MBA) as an initiator and crosslinker, respectively. The absorbency in various chloride salt solutions indicated that the absorbency decreased with increasing ionic strength of the salt solutions. Moreover, the absorbency under load (AUL) of SAPC was investigated at various applied loading and results show that, AUL decreased with increasing applied loading. Infrared Spectroscopy (IR) and Thermogravimetric Analysis (TGA) were carried out to confirm the chemical structure and thermal properties of the synthesized superabsorbent, respectively.
Protease was extracted from two maturity stages of noni fruits (Morinda citrifolia L.), unripe (stage 1) and ripe (stage 5). The crude extract was partially purified by acetone precipitation method followed by dialysis, gel filtration chromatography and freeze drying. Protein concentrations, proteolytic activity, molecular weight distribution, pH stability, temperature stability and storage efficiency of the resulting protease were evaluated. The unripe and ripe noni fruit contains 0.65 and 0.35% protein, respectively. Molecular weight of the proteases from both stages ranged approximately between 3 to 28 kDa based on the SDS-PAGE results. The optimum activity were at pH 7s and 6, temperatures of 40 and 50°C, respectively for proteases obtained from the unripe and ripe fruit. Analysis from the freeze dried protease indicated that protease from ripe noni fruits had higher protein concentration and specific activity compared to those from unripe fruit. However, it is more sensitive to pH and temperature and less stable during storage as it shows lower proteolytic activity compared to protease from unripe fruit. Based on its high proteolytic activity reaching up to 70.31 U/mg and storage stability (30% lost of activity), noni fruit could be an alternative source of plant protease.
Tomato (Solanum lycopersicum) is a globally important crop with an economic value in the tens of billions of dollars, and a significant supplier of essential vitamins, minerals, and phytochemicals in the human diet. Shelf life is a key quality trait related to alterations in cuticle properties and remodeling of the fruit cell walls. Studies with transgenic tomato plants undertaken over the last 20 years have indicated that a range of pectin-degrading enzymes are involved in cell wall remodeling. These studies usually involved silencing of only a single gene and it has proved difficult to compare the effects of silencing these genes across the different experimental systems. Here we report the generation of CRISPR-based mutants in the ripening-related genes encoding the pectin-degrading enzymes pectate lyase (PL), polygalacturonase 2a (PG2a), and β-galactanase (TBG4). Comparison of the physiochemical properties of the fruits from a range of PL, PG2a, and TBG4 CRISPR lines demonstrated that only mutations in PL resulted in firmer fruits, although mutations in PG2a and TBG4 influenced fruit color and weight. Pectin localization, distribution, and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to deesterified homogalacturonan, INRA-RU1 to rhamnogalacturonan I, LM5 to β-1,4-galactan, and LM6 to arabinan epitopes, respectively. The data indicate that PL, PG2a, and TBG4 act on separate cell wall domains and the importance of cellulose microfibril-associated pectin is reflected in its increased occurrence in the different mutant lines.
In this research, the physical, mechanical and morphological properties of oil palm empty fruit bunch (EFB) mat/woven kenaf fabric-reinforced epoxy composites have been investigated. The oil palm EFB/woven kenaf fabrics were varied, with weight ratios of 50/0 (T1), 35/15 (T2), 25/25 (T3), 15/35 (T4) and 0/50 (T5). The composites were fabricated using a simple hand lay-up technique followed by hot pressing. The result obtained shows that an increase in kenaf fiber content exhibited higher tensile and flexural properties. On the other hand, the opposite trend was observed in the impact strength of hybrid composites, where an increase in kenaf fiber content reduced the impact strength. This can be corroborated with the physical properties analysis, where a higher void content, water absorption and thickness swelling were observed for pure oil palm EFB (T1) composites compared to other samples. The scanning electron microscopy analysis results clearly show the different failure modes of the tensile fractured samples. Statistical analysis was performed using one-way ANOVA and shows significant differences between the obtained results.
Activated carbons can be produced from different precursors, including coals of different ranks, and lignocellulosic materials, by physical or chemical activation processes. The objective of this paper is to characterize oil-palm shells, as a biomass byproduct from palm-oil mills which were converted into activated carbons by nitrogen pyrolysis followed by CO2 activation. The effects of no holding peak pyrolysis temperature on the physical characteristics of the activated carbons are studied. The BET surface area of the activated carbon is investigated using N2 adsorption at 77 K with selected temperatures of 500, 600, and 700°C. These pyrolysis conditions for preparing the activated carbons are found to yield higher BET surface area at a pyrolysis temperature of 700°C compared to selected commercial activated carbon. The activated carbons thus result in well-developed porosities and predominantly microporosities. By using this activation method, significant improvement can be obtained in the surface characteristics of the activated carbons. Thus this study shows that the preparation time can be shortened while better results of activated carbon can be produced.
Fusarium species are known to cause various diseases on plantations including fruits and vegetables. The most common Fusarium that can cause plant diseases are Fusarium proliferatum and Fusarium verticillioides. Ear rot disease on maize, wilt disease on cucurbits and fruit rot disease on tomato as well as banana are example of diseases caused by these two species. The objectives of this study were to identify F. proliferatum and F. verticillioides based on species-specific primers and polymerase chain reaction (PCR) amplification and to evaluate the genetic diversity of both species based on microsatellite markers. Fifty isolates of Fusarium species that were previously collected throughout Malaysia from different hosts were identified by using species-specific PCR amplification. Twenty-nine isolates were identified as F. proliferatum and 21 isolates were identified as F. verticillioides based on species-specific primer. The genetic diversity of all the fungal isolates was evaluated by using microsatellite analysis with six established primers. Five out of six primers amplified polymorphic bands with the most effective primer showing high polymorphism were (AG)7C and (TCC)5 meanwhile one primer (TTTC)4 gave negative result with no band amplified. The phylogenetic tree that was constructed showing two different clades distinguished between F. proliferatum and F. verticillioides.
It is known from our previous studies that kiwifruits, which are used in common human diet, have preventive properties of coronary artery disease. This study describes a combination of (1)H NMR spectroscopy, multivariate data analyses and fluorescence measurements in differentiating of some kiwifruit varieties, their quenching and antioxidant properties. A total of 41 metabolites were identified by comparing with literature data Chenomx database and 2D NMR. The binding properties of the extracted polyphenols against HSA showed higher reactivity of studied two cultivars in comparison with the common Hayward. The results showed that the fluorescence of HSA was quenched by Bidan as much as twice than by other fruits. The correlation between the binding properties of polyphenols in the investigated fruits, their relative quantification and suggested metabolic pathway was established. These results can provide possible application of fruit extracts in pharmaceutical industry.
Olive fiber is a renewable natural fiber which has potential as an alternative biomass for extraction of microcrystalline cellulose (MCC). MCC has been widely applied in various industries owing to its small dimensional size for ease of reactive fabrication process. At present study, a serial treatments of bleaching, alkaline and acid hydrolysis was employed to extract OL-BLF, OL-PUF, and OL-MCC respectively from olive stem fiber. In morphology examination, a feature of short micro-crystallite particles was obtained for OL-MCC. The particle size was found gradually reducing from OL-PUF (305.31 μm) to OL-MCC (156.06 μm) due to the disintegration of cellulose fibrils. From physicochemical analysis, most lignin and hemicellulose components had been removed from OL-BLF to form OL-PUF with individually fibril structure. The elemental analysis revealed that highly pure cellulose component was obtained for OL-MCC. Also, the rigidity had been improved from OL-BLF to OL-PUF, while with the highest for OL-MCC with 74.2% crystallinity, endowing it as a reliable load-bearing agent. As for thermal analysis, OL-MCC had the most stable heat resistance in among the chemically-treated fibers. Therefore, olive MCC could act as a promising reinforcing agent to withstand harsh conditions for variety fields of composite applications.
The present work studies the profiling of phenolic bioactive and in vitro biological (anticancer, antioxidant, and antimicrobial) activities of different solvent extracts from Withania
somnifera fruit. Anticancer activity was performed using potato-disc assay and Agrobacterium tumefaciens. While antibacterial and antifungal evaluation was done by using disc diffusion method against bacterial (Staphylococcus aureus, S. epidermidis, Escherichia coli, and
Klebsiella pneumonia) and fungal (Aspergillus flavus and Fusarium oxysporum) strains.
Among different extraction solvents used, n-hexane extract exhibited the highest inhibition of
tumour initiation (64%), whereas ethyl acetate (15%) was the lowest by using potato-disc
assay. Highest total phenolic and total flavonoid contents were noted for methanolic (69.10
GAE mg/g DW%) and n-hexane (29.45 CE mg/g DW%) extracts, respectively. For antioxidant potential, 2,2,1-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC50) and reducing power EC50 were noted to be superior (0.6 and 2.0 mg/mL, respectively) for n-hexane
extract. All the tested extracts showed considerable antibacterial and antifungal activity with
the highest growth inhibition zones for K. pneumoniae (31.70 mm) and A. flavus (27.09 mm)
were shown by n-hexane extract. High Performance Liquid Chromatographic (HPLC) analysis of individual phenolics (gallic acid, 2,288.48 mg/kg) indicated the highest contents of these
compounds in n-hexane extract, which might explain the potent biological activities of this
extract. Our findings revealed that the bioactive present in the tested fruit had significant
potential as anticancer, antibacterial, and antifungal agents. Further studies are needed to
elucidate the mechanism of actions of isolated bioactive against specific diseases such as
cancer, especially in the case of n-hexane fraction.
The Baobab (Adansonia digitata L.) is a large iconic tree indigenous to Africa where it is found in many countries. The Baobab tree has various uses, as it produces food and non-food products such as medicines, fuel, timber and fodder. This research is focused on the characterization of the Baobab fruit shells in terms of lignin (54.08%), cellulose (24.87%) and hemicellulose (21.05%) content, as well as proximate analysis such as ash content (5.17%), moisture content (6.48%), volatile matter (86.73%) and carbon content (1.22%). This assessment will play a vital role in exploring the benefits of utilizing baobab fruit shells in the production of activated carbon as well as set a foundation for future research.
The present study was carried out to characterize red fruit (Pandanus conoideus Lam) oil (RFO) in term of FTIR spectra, fatty acid composition, and volatile compounds. FTIR spectrum of RFO was slightly
different from other common vegetable oils and animal fats, in which in the frequency range of 1750 – 1700 cm-1, RFO appear two bands. The main fatty acid composition of RFO is oleic acid accounting for 68.80% followed by linoleic acid with the concentration of 8.49%. The main volatile compounds of RFO as determined using gas chromatography coupled with mass spectrometry (GC-MS) and headspace analyser are 1,3-dimethylbenzene (27.46%), N-glycyl- L-alanine (17.36%), trichloromethane (15.22%), and ethane (11.43%).
The fundamental trait in selective breeding of oil palm (Eleais guineensis Jacq.) is the shell thickness surrounding the kernel. The monogenic shell thickness is inversely correlated to mesocarp thickness, where the crude palm oil accumulates. Commercial thin-shelled tenera derived from thick-shelled dura × shell-less pisifera generally contain 30% higher oil per bunch. Two mutations, sh MPOB (M1) and sh AVROS (M2) in the SHELL gene - a type II MADS-box transcription factor mainly present in AVROS and Nigerian origins, were reported to be responsible for different fruit forms. In this study, we have tested 1,339 samples maintained in Sime Darby Plantation using both mutations. Five genotype-phenotype discrepancies and eight controls were then re-tested with all five reported mutations (sh AVROS , sh MPOB , sh MPOB2 , sh MPOB3 and sh MPOB4 ) within the same gene. The integration of genotypic data, pedigree records and shell formation model further explained the haploinsufficiency effect on the SHELL gene with different number of functional copies. Some rare mutations were also identified, suggesting a need to further confirm the existence of cis-compound mutations in the gene. With this, the prediction accuracy of fruit forms can be further improved, especially in introgressive hybrids of oil palm. Understanding causative variant segregation is extremely important, even for monogenic traits such as shell thickness in oil palm.
The pretreatment of empty fruit bunch (EFB) was conducted using an integrated system of IL and cellulases (IL-E), with simultaneous fermentation in one vessel. The cellulase mixture (PKC-Cel) was derived from Trichoderma reesei by solid-state fermentation. Choline acetate [Cho]OAc was utilized for the pretreatment due to its biocompatibility and biodegradability. The treated EFB and its hydrolysate were characterized by the Fourier transform infrared spectroscopy, scanning electron microscopy, and chemical analysis. The results showed that there were significant structural changes in EFB after the treatment in IL-E system. The sugar yield after enzymatic hydrolysis by the PKC-Cel was increased from 0.058 g/g of EFB in the crude sample (untreated) to 0.283 and 0.62 ± 06 g/g in IL-E system after 24 and 48 h of treatment, respectively. The EFB hydrolysate showed the eligibility for ethanol production without any supplements where ethanol yield was 0.275 g ethanol/g EFB in the presence of the IL, while lower yield obtained without IL-pretreatment. Moreover, it was demonstrated that furfural and phenolic compounds were not at the level of suppressing the fermentation process.
Flour was prepared from peeled and unpeeled banana Awak ABB. Samples prepared were subjected to analysis for determination of chemical composition, mineral, dietary fibre, starch and total phenolics content, antioxidant activity and pasting properties. In general, flour prepared from unpeeled banana was found to show enhanced nutrition values with higher contents of mineral, dietary fibre and total phenolics. Hence, flour fortified with peel showed relatively higher antioxidant activity. On the other hand, better pasting properties were shown when banana flour was blended with peel. It was found that a relatively lower pasting temperature, peak viscosity, breakdown, final viscosity and setback were evident in a sample blended with peel.