Phenolics, their antioxidant and antimicrobial activity in dark germinated fenugreek sprouts in response to peptide and phytochemical elicitors.

The phenylpropanoid pathway (PPP) was stimulated in fenugreek sprouts through the pentose phosphate and shikimate pathway, by natural elicitors such as Fish Protein Hydrolysates (FPH), Lactoferrin (LF) and Oregano Extract (OE). Among treatments 0.5 ml/L FPH elicited fenugreek sprouts had the highest phenolic content of 0.75 mg/g FW on day 3 of germination which was approximately 25 % higher than control on the same day. The antioxidant activity estimated by beta-carotene assay was highest for LF and OE elicited sprouts on day 2 and 4, respectively with an antioxidant protection factor (APF) of 1.47 for both. In all treatments and control, higher antioxidant activity was observed during early germination, which correlates to higher phenolic content, suggesting that initially phenolics are antioxidant in nature. This increased activity also correlates with high guaiacol peroxidase (GPX) activity indicating that the polymerized phenolics required for lignification with growth have antioxidant function. The antioxidant activity as estimated by beta-carotene and 1,1,-diphenyl-2-picryl hydrazyl (DPPH) assays indicate that fenugreek sprout extract can quench the superoxide free radical and also possibly scavenge the hydrogen peroxide generated in the reaction mix. OE elicited the highest levo dihydroxy phenylalanine (L-DOPA) synthesis of 1.59 mg/g FW, followed by FPH with 1.56 mg/g FW and LF 1.5 mg/g FW all on day 2 which was 24.5%, 23 % and 20 % higher than control, respectively. Higher L-DOPA content was observed in the elicited fenugreek sprouts during early germination, correlating to high phenolics and antioxidant activity, suggesting that L-DOPA also contributes to the high antioxidant activity. The glucose-6-phosphate dehydrogenase (G6PDH) activity was higher during early germination (day 1-4) and gradually decreased during later stages (day 5-8) for all treatments and control. The early increase is possibly due to the carbohydrate mobilization from the cotyledons directed towards the high nutrient requirements of the growing sprout. As mobilization occurred, an allosteric feedback inhibition by sugar-phosphates is suggested, as lower G6PDH activity was observed on days 6-8. The elevated levels of GPX during early germination coincide with the higher phenolic synthesis; SOD activity and antioxidant activity suggests the elevated production and quenching of reactive oxygen species by elicitation. High antimicrobial activity against peptic ulcer-linked Helicobacter pylori was observed in the fenugreek sprout extract from control and LF treatments only. We hypothesized that in fenugreek sprouts, simple free phenolics that are less polymerized have more antimicrobial function.