Strain improvement of Sporidiobolus johnsonii -ATCC 20490 for biotechnological production of Coenzyme Q10

Ranadive, Prafull; Mehta, Alka; George, Saji

doi:10.7763/ijcea.2011.v2.106

Cited by 10 publications

(18 citation statements)

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“…Mutant EA22 that was previously generated from wild type strain S. johnsonii ATCC 20490 [17] was used as a starting strain for further improvement. Natural variant of this mutant, E2A22 was selected on YM agar plate, tested in shake flask and further re-mutated using combined exposure of UV and free radicals.…”

Section: Mutagenesis and Selection Of Mutantsmentioning

confidence: 99%

“…In our previous study, an ethyl methane sulfonate (EMS) induced (exposure at 20 ll ml -1 for 30 min with 92 % reduction in viability), atorvastatin (20 lg ml -1 ) tolerating mutant EA22 was generated from S. johnsonii ATCC-20490 with 2-fold improvement in CoQ 10 titer (HPTLC method) [17]. EA22 showed mycelial growth forms like wild type strain but it was less pigmented and had reduced cell size.…”

Section: Mutagenesis Selection Of Mutants and Screening In Shake Flaskmentioning

confidence: 99%

“…In the present study, a natural variant of this mutant (E2A22) that showed maximum 1.6-fold improvement in specific CoQ 10 content was selected and used as a starting strain for further improvement. To overcome the limitations of HPTLC method for CoQ 10 estimation, in terms of accuracy when used for primary screening of mutants [17], the HPLC method was followed for subsequent screenings. E2A22 was subjected to combined exposure of UV and free radicals generated by photodynamic action of methionine-riboflavin (MR) mixture [38].…”

Section: Mutagenesis Selection Of Mutants and Screening In Shake Flaskmentioning

confidence: 99%

“…The isolation of CoQ 10 from S. johnsonii has been reported by one group, using improved fermentation process involving feeding of para-hydroxy benzoic acid, a precursor of benzoquinone moiety of CoQ 10 [16]. This strain has not been explored extensively for CoQ 10 studies like few higher producing bacterial strains Agrobacterium tumifaciens (Rhizobium radiobacter) and Rhodobacter sphaeroides [2] due to its low CoQ 10 content, hence the strain improvement work on this strain was undertaken by us [17]. Improving the yields of CoQ 10 in S. johnsonii would be beneficial so as to take the advantage of having more biomass production in yeast fermentation.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, yeasts like S. johnsonii becomes a promising source of natural CoQ 10 derived from eukaryote kingdom having high degree of conservation with mammalian system and its biosynthetic pathways [18]. During strain improvement program on this strain, the improved mutant strain EA22 was previously generated by EMS mutagenesis followed by rational selection [19] on atorvastatin (20 lg ml -1 ) [17]. Re-mutagenesis process brings about improvement in yields [20]; hence we continued our efforts on this strain to bring significant improvement in CoQ 10 content by induced re-mutagenesis and rational selection.…”

Section: Introductionmentioning

confidence: 99%

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Morphological and Molecular Differentiation of Sporidiobolus johnsonii ATCC 20490 and Its Coenzyme Q10 Overproducing Mutant Strain UF16

et al. 2014

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Coenzyme Q 10 (CoQ 10 ) is an industrially important molecule having nutraceutical and cosmeceutical applications. CoQ 10 is mainly produced by microbial fermentation and the process demands the use of strains with high productivity and yields of CoQ 10 . 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 CoQ 10 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 rRNAgene 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 CoQ 10 biosynthetic pathway that resulted in improved yield. Hence, investigating the mutations responsible for deregulation of CoQ 10 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 CoQ 10 production.

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Section: Mutagenesis and Selection Of Mutantsmentioning

confidence: 99%

Section: Mutagenesis Selection Of Mutants and Screening In Shake Flaskmentioning

confidence: 99%

Section: Mutagenesis Selection Of Mutants and Screening In Shake Flaskmentioning

confidence: 99%