The tentative identification of bioactive compounds in the extract of Vernonia amygdalina leaf was carried out using positive ionization of Liquid chromatography-mass spectrometry quadrupole time of flight (LC-Q-TOF/MS). The positive ionization is associated with the presence of saponins, flavonoids, alkaloids, terpenoids, and glycosides. Tentative assignments of the secondary metabolites were performed by comparing the MS fragmentation patterns with Waters® UNIFY library which allows positive identification of the compounds based on the spectral match. All the metabolites compounds were estimated and presented in a BPI (Base peak intensity) plot. These data are the unpublished supplementary materials related to “Ethanolic extraction of bioactive compounds from V. amygdalina leaf using response surface methodology as an optimization tool” (Alara et al., 2018).
The saccharification of laccase-pretreated empty fruit bunch (EFB) was optimized in a lab-scale experiment using one-factor-at-a-time (OFAT) and response surface methodology (RSM). After pretreatment, the degree of delignification was checked by noting the weight loss (%) after pretreatment, and also by the quantity of total sugar produced after saccharification with cellulase enzyme. OFAT studies of saccharification of the pretreated EFB showed that the biomass was best saccharified using cellulase enzyme at the following conditions: enzyme concentration of 30 IU/g of EFB, substrate concentration of 5.0% w/v, 50 °C, saccharification time of 24 h, and pH 5. This combination exhibited the highest yield of total sugar (28% w/w). Although 29% w/w yield was achieved with an enzyme concentration of 40 IU/g of EFB, this increase in yield was not proportional to the increased enzyme concentration and, therefore, was considered insignificant. Statistical analysis of the combined effects of pH and temperature showed that pH had a more significant effect than the temperature on the saccharification process, based on a P < 0.05 significance level. The effect of pH on total sugar production was more significant than the temperature in both linear and quadratic functions. In sum, the saccharification of laccase-pretreated EFB should follow the optimized process conditions achieved in the current study.
Laccase enzyme was used as a pretreatment agent to delignify empty fruit bunches (EFB) for sugar production. The degree of delignification of the biomass was assessed directly by the percentage of pre-pretreatment weight loss (%) after pretreatment and indirectly by the amount of total sugar produced after saccharification of the pretreated biomass with cellulase enzymes. Process parameters such as pretreatment time, temperature, enzyme concentration, substrate concentration, pH, and substrate size were studied using a one-factor-at-a-time (OFAT) analysis. The combined effect of temperature and pH on the pretreatment was studied using the face-centered central composite design (FCCCD) of response surface methodology (RSM). The optimized conditions for EFB pretreatment using laccase enzyme were achieved as follows: sample size, 2 mm; temperature, 25 °C; time, 4 h; substrate concentration, 5% (w/v); pH 5; and enzyme concentration, 20 IU/g of EFB. Although higher pretreatment was achieved with substrates of 1 mm size and at a temperature of 35 °C, these conditions were not considered energetically sustainable because of the need for energy during milling for sample size reduction and energy for temperature maintenance at 35 °C.
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