Screening and characterization of lipase producing bacteria isolated from the gut of a lepidopteran larvae, Samia ricini

Abstract

• Rhodamine B screening identified twenty-eight lipolytic gut bacterial isolates. • Majority of lipolytic bacteria belonged to the genus Bacillus and Pseudomonas. • Isolates ERI011 and ERI080 from the foregut had the highest lipase activity. • S. ricini could be a resource for prospecting lipolytic microbes for use in biotechnology. Lipolytic enzymes are an important group of hydrolases that have found immense industrial application in biotechnology. In this study, the ability of gut bacteria isolated from the gut of the Eri silkworm, Samia ricini, to produce lipolytic enzymes was evaluated through qualitative and quantitative assays. The results of lipase screening showed that 28 isolates had lipolytic activity. The results of 16S ribosomal RNA sequencing indicated that the genus Bacillus comprised majority of the lipolytic bacterial isolates (71%) followed by Pseudomonas (15%); whilst Acinetobacter, Enterobacter and Enterococcus comprised 11%. Lipolytic activity was found in bacteria isolates identified from all the three gut compartments of S. ricini larvae with significant activity from isolates extracted from the foregut and midgut. The lipolytic index among the bacterial isolates ranged between 0.63 and 2.81 on Rhodamine B medium, and all isolates exhibited significant lipolytic activity with p -nitrophenyl butyrate (PNPB) with specific activity ranging from 0.52 to 0.82 μmol/min/mg. The effect of pH and temperature showed that lipase activity was optimum at 37 °C and pH 7–9. A phylogenetic relationship of lipase producing gut bacteria indicated high cluster stability for isolates from different stages (>50%) suggesting that the isolates persist across developmental stages of the host. The Eri silkworm is reared for its silk and the knowledge of its gut bacteria with the ability to produce lipases lies in the significance as far as boosting production of this insect via development of probiotics to enhance commercial Eri rearing. In addition, this insect may be a good resource for profiling novel lipolytic microbes for commercial production of lipases as lipases from microbial origin have assumed a great deal of importance as industrial enzymes due to their potential for use in biotechnology.