Regional Symposium on Innovation in science and technology

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Targeted Identification of biopolymer sphingan producers: RAPD-PCR analysis and the development of degenerate PCR primers

Authors :

Monir-sadat Shakeri¹

1- Research Institute of Food science And Technology

Keywords :

Sphingomonas identification،Cluster analysis،RAPD-PCR،MCA

Abstract :

Sphingomonas strains are the main focus for producing various microbial polysaccharides such as welan, gellan, and diutan, all of which are based on sphingan. These substances and their different forms are commonly used in various fields, including medicine, food, and the petroleum industry. It is crucial to identify local strains in order to obtain high-yield strains. In this study, screening of indigenous Sphingomonas isolates obtained from environmental sources including soil and water was conducted using degenerate primers designed from the pgmG gene, along with RAPD-PCR technique. The strains were precisely identified by sequence analysis of the 16SrDNA region. The study found 44 bacterial isolates classified as gram-negative bacilli that showed positive results for the catalase test. The isolates were analyzed for RAPD patterns, which led to the identification of 14 subclusters with a similarity of 45%. The identification of ten distinct Sphingomonas strains was achieved through sequencing the 16SrDNA within these subclusters. The designed primers were assessed to determine the target band, revealing that all ten strains produced the expected band and have the ability to produce sphingan-based biopolymers. Combining PCR with degenerate primers and RAPD analysis provides a systematic and accurate method for carrying out epidemiological research on sphingomonas strains