Recombinant Expression and Enzyme Activity of Chymotrypsin-like Protease from Black Soldier Fly, Hermetia illucens (Diptera: Stratiomyidae)

Abstract

Chymotrypsin serine protease is one of the main digestive proteases in the midgut of and is involved in various essential processes. In a previous study, a gene encoding a chymotrypsin-like protease, Hi-SP1, was cloned from the larvae of Hermetia illucens and characterized. In this study, we produced the recombinant chymotrypsin-like protease Hi-SP1 in Escherichia coli cells. The molecular weight of the recombinant Hi-SP1 was estimated to be approximately 26 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western-blotting. Chymotrypsin activity was detected when AAPF was used as the substrate. Examination of the effects of temperature and pH revealed that the proteolytic activity of recombinant Hi-SP1 decreased markedly at temperatures above 30 o C, and the optimum pH was found to be 10.0.

Rethinking organic wastes bioconversion: Evaluating the potential of the black soldier fly (Hermetia illucens (L.)) (Diptera: Stratiomyidae) (BSF)

“…Moreover, economicallyimportant bioproducts such as protein hydrolysates (Firmansyah and Abduh, 2019), bioplastic (Barbi et al, 2019), and natural pigments (e.g., melanin and ommochromes) (Ushakova et al, 2019) could also be derived from BSF biomass. Studies have also explored BSF biomass as a potential source of industrially important enzymes (e.g., trypsin, chymotrypsin, ligninase, and cellulase) (Bonelli et al, 2019;Kim et al, 2011;Lee et al, 2014;Muller et al, 2017;Park et al, 2012) and antimicrobial peptides. The successful development of a BSF-based biorefinery, however, will require a large-scale production of BSF biomass, which in turn will require mechanization and automation of rearing, harvesting, and processing.…”

The black soldier fly, Hermetia illucens – a promising source for sustainable production of proteins, lipids and bioactive substances

“…Since 1977 the concentration for erucic acid in food is not allowed to exceed 5% in Germany [33]. trypsin and chymotrypsin activity has been published [34,35]. In 2014, Lee et al furthermore heterologously expressed a novel cellulase gene from the gut microflora and characterized the resulting enzyme which shows 72% identity and 80% homology to the glycosyl hydrolase family 5 of Dysgonomonas mossii [36].…”

Structural and Functional Characterization of Hermetia illucens Larval Midgut

“…Moreover, a qualitative and quantitative comparison of the digestive enzymatic activity from M. domestica and H. illucens larvae demonstrated that the latters possess more digestive enzymes with higher levels of activity ( Kim et al, 2011b ), a finding that supports the extraordinary digestive capability of H. illucens . Finally, two serine proteases were cloned and characterized ( Kim et al, 2011a ; Park et al, 2012 ).…”