Bioactives from plants are always in high demand in nutraceutical, pharmaceutical and functional food sectors due to their health benefits. This intensifies the need of extraction of bioactives by different methods that can improve the yield and purity of the compound. Enzyme helps in the release of bioactives from the plant material under optimized conditions so as to make the extraction process efficient. Though enzymatic extraction of bioactive has been used since long, it needs improvement to further enhance the yield, reduce the process time and to make the process cost competitive. The combination of enzymatic extraction with other green techniques such as ultrasound extraction, supercritical fluid extraction, three phase partitioning, ionic liquid extraction and microwave extraction can boost the advantages of enzymatic extraction. This review focuses on the improvement in the enzymatic extraction techniques for the bioactive in detail. Keywords Bioactives Enzyme-assisted extraction (EAE) Ultrasound assisted enzyme extraction (UAEE) Enzyme-assisted supercritical extraction (EASCFE) Ionic liquids enzyme assisted extraction (ILEAE) Microwave assisted enzymatic extraction (MAEE) Enzyme assisted three phase partition (EATPP) High pressure assisted enzymatic extraction (HPAEE)

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Summer truffles (Tuber aestivum) collected from Italy were evaluated for changes in chemical composition and hence in vitro bioactivity on the basis of their maturity. Truffles were classified as immature and mature based on the colour of glebum. Water and methanol extracts of freeze-dried mature truffles demonstrated higher total phenolics, higher in vitro antioxidant activity and lower tannins than immature truffle extracts. Phenolics and flavonoids were detected by Liquid Chromatography-Mass spectroscopy (LC-MS) analysis. Gas chromatography (GC)-MS analysis of hexane extracts showed differences in fatty acid composition. Linoleic acid was highest in both samples, lauric and myristic acids were prominently detected in mature truffle hexane extracts. The aroma profiling of truffles by headspace-GC showed increase in alcohols and decrease in aldehydes. Thus, this study comprehends altered chemical composition and its manifestation in in vitro bioactivity on maturation of summer truffles. This would aid development of techniques for identification of maturity of truffles.