Optimization of jacalin extracted from jackfruit (artocarpus heterophyllus) seed using reverse micellar extraction

Jacalin has excellent potentials in improving the therapeutic efficacy of anticancer drugs, especially in the diagnosis and active targeting of cancer cells. However, the widespread application was limited by the conventional purification process that was expensive, time-consuming and difficult to scale up. The aim of this study was to investigate the application of reverse micellar extraction (RME) consisting of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) reverse micelles in isooctane for the recovery and purification of jacalin from the jackfruit seed crude extract. For the optimization of forward extraction, one-factor-at-a-time (OFAT) approach and fractional factorial design (FFD) was first employed to determine the significant factors affecting the forward extraction efficiency (FEE). The FFD determined that the aqueous phase pH, NaCl concentration and AOT concentration were the significant factors and were optimized using Box-Behnken design (BBD). The maximum FEE of 88.04±1.30% was obtained at aqueous phase pH 4.58, 125 mM NaCl and 40 mM AOT. For backward extraction, the effects of stripping aqueous phase pH, KCl concentration and isopropyl alcohol (IPA) on backward extraction efficiency (BEE) were evaluated based on OFAT experiments followed by optimization using BBD. The maximum BEE was 92.60 ± 0.84 % at pH 9.0, 0.75M KCl and 40 %v/v IPA. The partitioning kinetic and isotherm at the optimized conditions of forward extraction were examined using pseudo first order (PFO), pseudo second order (PSO), Langmuir and Freundlich models. The results revealed that the optimized forward extraction data fit well with the PSO model (R2 = 0.9829) than the PFO model (R2=0.9738). Isotherm studies indicated that the Langmuir isotherm model best fit the data with a high R2 value (0.9315) than the Freundlich (R2=0.8728). Mass transfer mechanism based upon the two-film theory revealed that the range of combined mass transfer coefficients obtained is 0.2335 × 10-8 m3/s to 0.7940 × 10-8 m3/s for forward extraction and 0.1852 × 10-8 m3/s to 0.3479 × 10-8 m3/s for backward extraction. For the effect of pH, the forward extraction was limited by the diffusion in the reverse micellar phase, while the diffusion resistance in the stripping aqueous phase controls the backward extraction. Zebrafish embryos treated with different concentration of the jackfruit seed crude extract and extracted jacalin showed embryonic and developmental defects. The 96hr-LC50 of jackfruit crude seed extract and jacalin were 20.48 μg/ml and 14.81 μg/ml, respectively. Multiple developmental abnormalities such as pericardial edema, spinal curvature, decreased heart rate and delayed hatching were observed in a concentration-dependent manner. Overall, this study indicated that the AOT/isooctane reverse micellar system is an efficient method to extract and purify jacalin from jackfruit seeds with maximum extraction efficiency. The extracted jacalin, however, could pose threats when consumed at higher doses especially on embryos. In the future, the RME method may be exploited for the recovery and purification of similarly high-value proteins from complex biological mixtures. However, detailed toxicity analysis on the extracted proteins should be performed to ascertain their safety on the developing embryos

The Susceptibility of Aphids, Aphis gossypii Glover to Lauric Acid Based Natural Pesticide

The susceptibility of aphids to lauric acid, a type of middle-chain fatty acids, based pesticide formulation was investigated. The insecticidal properties of lauric acid based pesticide formulation against aphids were evaluated through the three main routes of exposure: topical application, leaf-dipping (ingestion) and inhalation. Surface tension dependent aphids toxicity was also investigated by measuring the surface tension value of each formulation prepared. The relationship of surface tension and formulation toxicity was then observed on aphid tested. The obtained results indicate that all tested formulations had insecticidal activity against tested aphids depending on exposure method and lauric acid concentration. The topical application method recorded 100% mortality rates compared to the other two methods. With topical application method, most of the aphids died soon after treatment. Toxicity symptoms from lauric acid to aphids included unusual abdominal contractions which caused quick loss of coordination and decreased activity, and finally dehydration and necrosis within 24 hours. Results from surface tension study revealed close connection between toxicity and surface tension. This might lead to the conclusion that toxicity was possibly related to aphid suffocation in the highly wettable solution. Results from this research also demonstrated that there were possibility that lauric acid do not only kill aphids by suffocation, but also through change of cuticle and cell permeability and desiccation that lead to speedy death due to the chemical nature of lauric acid. Overall, the results recommend that lauric acid based pesticide formulation has both physical and chemical mode of action on aphids

Development and characterization of bio-composite films made from bacterial cellulose derived from oil palm frond juice fermentation, chitosan and glycerol

This study reported for the first time, the combined effects of chitosan and glycerol addition on the properties of bacterial cellulose (BC) based films for food packaging applications. Films were prepared by solution casting method using BC derived from oil palm frond juice as the main material combined with different concentrations of chitosan (0.5 and 1 %w/v) and glycerol (0.5, 1.5 and 2.5 %v/v). Pure BC, chitosan-free and glycerol-free films were used as control. The effect of incorporating chitosan and glycerol on bacterial cellulose (BC) based films was evaluated based on the physical properties (thickness, moisture content, solubility), mechanical properties (tensile strength, modulus Young, elongation at break) and chemical structure by FTIR. Increased concentration of chitosan and glycerol affected the physical and mechanical properties. The combination of 1 %w/v chitosan and 0.5 %v/v glycerol had a strengthening effect on the BC-based films with maximum tensile strength of 15 MPa and Young’s modulus of 772 MPa. Meanwhile, BC films incorporated with 1 %w/v chitosan and 2.5 %v/v glycerol demonstrated high plasticizing effect of 7 % elongation at break. The acquired FTIR spectrum of the bio-composite films suggested intermolecular interactions between BC, chitosan, and glycerol. Therefore, the BC-based bio-composite films incorporated with chitosan and glycerol have the potential to be used as food packaging materials

Lignin content analysis in oil palm frond juice base medium: effect on bacterial cellulose production by Acetobacter xylinum 0416

Extensive work on bacterial cellulose (BC) cultivation is crucial in optimizing BC performance and versatility in application. Herein, this study aims to investigate the lignin content in oil palm frond (OPF) juice medium that could affect the BC production by Acetobacter xylinum 0416 in static culture. The effects of lignin content were observed by the performances of BC production from the raw and pre-treated OPF juice medium fermentation with Hestrin-Schramm (HS) medium as a point of comparison. The TAPPI methods analysis outlined up to 3.719 g/L of total lignin content in raw OPF juice and FTIR analysis depicted the functional group of a typical lignin compound, thus confirming its presence as a component inhibitor. The raw OPF juice was further pre-treated using an alkaline precipitation method resulting in acid-insoluble lignin removal up to 75.47% of the total lignin content. The results revealed that the BC yield from the raw OPF juice fermentation was lower compared to the HS medium with dried BC content of 0.152 g/L and 2.52 g/L, respectively. The BC cultivation using the pre-treated OPF juice medium was improved as the BC yield significantly increased to 1.91 g/L. Therefore, a proper pre-treatment strategy on OPF juice medium for lignin content removal could improve the medium capability in BC cultivation for higher-scale production

Factors influencing forward extraction of jacalin from aqueous phase into anionic reverse micelles

Background: This study deals with the extraction of jacalin from crude extract of jackfruit seed using anionic surfactant based reverse micellar system, dioctyl sulfosuccinate sodium salt (AOT) in isooctane. The effect of AOT concentration, NaCl concentration and aqueous phase pH on the forward extraction efficiency of jacalin was evaluated using one factor at a time experiments (OFAT). Results: AOT concentration, NaCl concentration and aqueous phase pH were found to have different effects on the removal of jacalin from the aqueous phase into the AOT reverse micelles. A maximum forward extraction efficiency of 78% was achieved after the mixture was stirred for 15 min using 10 mM AOT at pH 5 aqueous phase containing 0.1 M NaCl. Conclusion: The findings demonstrated AOT reverse micellar system as a promising and effective method to extract and purify jacalin from crude protein mixture

Ex-situ development and characterization of composite film based on bacterial cellulose derived from oil palm frond juice and chitosan as food packaging

The development of alternative food packaging films using bio-based residues is in great demand for replacing petroleum-based packaging materials. However, large-scale application is severely limited by costly production and poor performance. This study investigates the ex-situ modification of bacterial cellulose (BC) produced by Acetobacter xylinum in oil palm fronds juice to obtain BC-Chitosan (BCC) films. FTIR revealed the structure of amide I and II bands, confirming the presence of chitosan in BCC films. The FE-SEM images of BCC films showed the formation of a thick chitosan layer with increasing chitosan incorporated into the BC surface structure. The coated chitosan layer observed improved mechanical properties in BCC films due to the disappearance of empty pores between BC fibers. Increments in chitosan concentration slightly decreased the thermal behavior of BCC. The antimicrobial effects of BCC films were effective against Gram-positive bacteria (Staphylococcus aureus) when the concentration of chitosan incorporated was above 0.6 %w/v. This study reveals the potential of extending the application of BC derived from oil palm frond juice (OPFJ) for developing food packaging materials

Response surface optimization of the forward extraction of jacalin from jackfruit seeds using AOT/isooctane reverse micellar system

Jacalin is the major protein contained in the crude extract of jackfruit (Artocarpus heterophyllus) seed that specifically recognizes and binds reversibly to galactose. Conventionally, purification of jacalin is carried out using the tedious and costly chromatographic techniques. In this study, extraction of jacalin from jackfruit seed crude extract were done using the sodium bis(2-ethylhexyl) sulfosuccinate (AOT)-based reverse micellar system. Reverse micellar extraction is an attractive alternative for downstream processing of various proteins. A successful reverse micellar extraction consists of two basic steps: forward and backward extraction. Forward extraction transfers a target protein from an aqueous solution into the reverse micellar solution, while backward extraction releases the protein from the reverse micelles structure into a new aqueous solution. The effects of the aqueous phase pH, NaCl concentration and AOT concentration on the forward extraction efficiency (FEE) are investigated using the response surface methodology (Box-Behnken Design). The main effects and interactions of the parameters are analyzed through the 3D surface plots. The optimum conditions for forward extraction were determined as follows: aqueous phase pH 4.58, 125 mM NaCl and 40 mM AOT. Under the optimal conditions, the FEE reached 88.04±1.30%, closer to 87.99% predicted by the model. The results indicated that AOT/isooctane reverse micelle system is effective in extracting jacalin from the jackfruit seed crude extract and verified the practicability of the BBD model for optimizing the main parameters in the forward extraction of jacalin

Study of Essential Oil from Malaysian Curry Leaves

Essential oils of Murraya koengii or commonly known as curry leaves’ obtained from curry trees grown in the local area of Malaysia was studied. Conventional hydro distillation process was used to extract the oil from the leaves. The highest yield from the extraction is to be at 0.22% for 9 hours of extraction time. The analysis was performed using GC-MS which detects the presence of 30 different components, majorly hydrocarbons. The result shows the existing of alpha-pinene and beta-myrcene that provides insight to the potential of using the oil as active ingredients for natural based insect repellent

Effect of Key Parameters on Jacalin Extraction from Aqueous Phase into Anionic Reverse Micelles

This study demonstrates the extraction of jacalin from crude extract of jackfruit seeds using anionic surfactant based reverse micellar system, sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in isooctane. Effects of various parameters, such as, NaCl concentration (0.05 – 1 M), pH of aqueous phase (pH 4 – 10), AOT concentration (5 – 150 mM), contact time (5 – 30 min) and phase volume ratio (0.5-5) on the transfer efficiency of jacalin was evaluated by changing one factor at a time (OFAT) while keeping the other parameters constant. A maximum of 83% of protein transfer was achieved after equal volume of organic and aqueous phase was stirred for 20 min using 20 mM AOT at pH 5 aqueous phase containing 0.1 M NaCl. The findings demonstrated AOT reverse micellar system as a promising and effective method to extract and purify jacalin from crude protein mixture