Molecular and phylogenetic analysis of inulinase-producing yeast isolated from nira siwalan (Borassus flabellifer) based on ITS sequences

Abstract

Inulinase (E.C. 3.2.1.7) is an enzyme capable hydrolyze inulin to be a fructose monomer, which is widely used in various fields, especially the food industry. This enzyme can be found in various plant species and can be produced by microorganisms such as bacteria, yeast, and fungi. Previous research has succeeded in isolating the potential yeast of inulinase from nira siwalan (Borassus flabellifer), named N1. This research purpose to identify molecularly the yeast inulinolytic N1 and its kinship with other species based on internal transcribed spacer (ITS) rDNA sequences. The stages in this research are yeast DNA extraction using the chelex method, DNA amplification with primers ITS 4 and ITS 5, electrophoresis, sequencing analysis, and construction of the phylogenetic tree. Phylogenetic trees are reconstructed using the neighbor-joining method based on their evolutionary relationship. The results of DNA isolation showed that the concentration of yeast DNA was 495.8 ng/ μl and purity of 2,12. The PCR product from the ITS fragment amplification produced a band size of ± 510 bp. The results of molecular identification and phylogenetic analysis showed that the N1 yeast isolate was closely related to Candida parapsilosis (MK638869.1) originating from India with a homology of 99.45%.