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

• Main Author: Sharifah Fathiyah, Sy Mohamad
• Format: Thesis
• Language: English
• Published: 2022
• Subjects: T Technology (General); TP Chemical technology
• Online Access: http://umpir.ump.edu.my/id/eprint/35924/1/Optimization%20of%20jacalin%20extracted%20from%20jackfruit%20%28artocarpus%20heterophyllus%29%20seed%20using%20reverse%20micellar.ir.pdf; http://umpir.ump.edu.my/id/eprint/35924/
• Institution: Universiti Malaysia Pahang

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.