title: Morphological and Molecular Differentiation of Sporidiobolus johnsonii ATCC 20490 and Its Coenzyme Q10 Overproducing Mutant Strain UF16.
abstract: Coenzyme Q10 (CoQ10) is an industrially important molecule having nutraceutical and cosmeceutical applications. CoQ10 is mainly produced by microbial fermentation and the process demands the use of strains with high productivity and yields of CoQ10. During strain improvement program consisting of sequential induced mutagenesis, rational selection and screening process, a mutant strain UF16 was generated from Sporidiobolus johnsonii ATCC 20490 with 2.3-fold improvements in CoQ10 content. EMS and UV rays were used as mutagenic agents for generating UF16 and it was rationally selected based on atorvastatin resistance as well as survival at free radicals exposure. We investigated the genotypic and phenotypic changes in UF16 in order to differentiate it from wild type strain. Morphologically it was distinct due to reduced pigmentation of colony, reduced cell size and significant reduction in mycelial growth forms with abundance of yeast forms. At molecular level, UF16 was differentiated based on PCR fingerprinting method of RAPD as well as large and small-subunit rRNA gene sequences. Rapid molecular technique of RAPD analysis using six primers showed 34 % polymorphic fragments with mean genetic distance of 0.235. The partial sequences of rRNA-gene revealed few mutation sites on nucleotide base pairs. However, the mutations detected on rRNA gene of UF16 were less than 1 % of total base pairs and its sequence showed 99 % homology with the wild type strain. These mutations in UF16 could not be linked to phenotypic or genotypic changes on CoQ10 biosynthetic pathway that resulted in improved yield. Hence, investigating the mutations responsible for deregulation of CoQ10 pathway is essential to understand the cause of overproduction in UF16. Phylogenetic analysis based on RAPD bands and rRNA gene sequences coupled with morphological variations, exhibited the novelty of mutant UF16 having potential for improved CoQ10 production.
authors.affiliation: Fermentation Technology Lab, Natural Products Department, Piramal Enterprises Limited, Nirlon Complex, Off Western Express Highway, Goregaon (East), Mumbai, 400063 India.
authors.forename: Prafull
authors.initials: P
authors.lastname: Ranadive
authors.affiliation: School of Bio Science and Technology, VIT University, Vellore, 632014 Tamil Nadu India.
authors.forename: Alka
authors.initials: A
authors.lastname: Mehta
authors.affiliation: geneOmbio Technologies Private Limited, Baner, Pune, 411045 Maharashtra India.
authors.forename: Yashwant
authors.initials: Y
authors.lastname: Chavan
authors.affiliation: Fermentation Technology Lab, Natural Products Department, Piramal Enterprises Limited, Nirlon Complex, Off Western Express Highway, Goregaon (East), Mumbai, 400063 India.
authors.forename: Anbukayalvizhi
authors.initials: A
authors.lastname: Marx
authors.affiliation: Fermentation Technology Lab, Natural Products Department, Piramal Enterprises Limited, Nirlon Complex, Off Western Express Highway, Goregaon (East), Mumbai, 400063 India.
authors.forename: Saji
authors.initials: S
authors.lastname: George
keywords: Atorvastatin; Coenzyme Q10; Free radical; Mutant; Mutation; RAPD; Sporidiobolus johnsonii; Strain improvement