title: Approaches towards the enhanced production of Rapamycin by Streptomyces hygroscopicus MTCC 4003 through mutagenesis and optimization of process parameters by Taguchi orthogonal array methodology.
abstract: The present research was conducted to define the approaches for enhanced production of rapamycin (Rap) by Streptomyces hygroscopicus microbial type culture collection (MTCC) 4003. Both physical mutagenesis by ultraviolet ray (UV) and chemical mutagenesis by N-methyl-N-nitro-N-nitrosoguanidine (NTG) have been applied successfully for the improvement of Rap production. Enhancing Rap yield by novel sequential UV mutagenesis technique followed by fermentation gives a significant difference in getting economically scalable amount of this industrially important macrolide compound. Mutant obtained through NTG mutagenesis (NTG-30-27) was found to be superior to others as it initially produced 67% higher Rap than wild type. Statistical optimization of nutritional and physiochemical parameters was carried out to find out most influential factors responsible for enhanced Rap yield by NTG-30-27 which was performed using Taguchi orthogonal array approach. Around 72% enhanced production was achieved with nutritional factors at their assigned level at 23 °C, 120 rpm and pH 7.6. Results were analysed in triplicate basis where validation and purification was carried out using high performance liquid chromatography. Stability study and potency of extracted Rap was supported by turbidimetric assay taking Candida albicans MTCC 227 as test organism.
authors.affiliation: Department of Biotechnology, National Institute of Technology Durgapur, Mahatma Gandhi Avenue, Durgapur, 713209, India.
authors.forename: Subhasish
authors.initials: S
authors.lastname: Dutta
authors.affiliation: Energy Research and Technology Group, CSIR-Central Mechanical Engineering Research Institute, Mahatma Gandhi Avenue, Durgapur, 713209, India.
authors.forename: Bikram
authors.initials: B
authors.lastname: Basak
authors.affiliation: Department of Bioengineering, National Institute of Technology Agartala, Barjala, Tripura, 799055, India.
authors.forename: Biswanath
authors.initials: B
authors.lastname: Bhunia
authors.affiliation: Department of Biotechnology, National Institute of Technology Durgapur, Mahatma Gandhi Avenue, Durgapur, 713209, India.|Bioprocess Development Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India.
authors.forename: Ankan
authors.initials: A
authors.lastname: Sinha
authors.affiliation: Department of Biotechnology, National Institute of Technology Durgapur, Mahatma Gandhi Avenue, Durgapur, 713209, India. apurbadey.bt@gmail.com.
authors.forename: Apurba
authors.initials: A
authors.lastname: Dey
mesh_terms: D005285:Fermentation; D007218:Industrial Microbiology; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D016296:Mutagenesis; D020123:Sirolimus / Q000378:metabolism*; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D008769:Methylnitronitrosoguanidine; D020123:Sirolimus
keywords: Nitrosoguanidine; Rapamycin; Sequential UV mutagenesis; Streptomyces hygroscopicus; Turbidimetric assay