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Gamma radiation induced mutagenesis in Aspergillus niger to enhance its microbial fermentation activity for industrial enzyme production

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Abstract

α- and β-Galactosidases find application in food processing, health and nutrition. Aspergillus niger is one of the potent producer of these enzymes and was genotypically improved using gamma-ray induced mutagenesis. The mutant-derivative produced two-fold higher α- and β-galactosidases. For testing genetic variability and its relationship with phenotypic properties of the two organisms, DNA samples of the mutant and parental strains of A. niger were amplified with 28 deca-nucleotide synthetic primers. RAPD analysis showed significantly different pattern between parental and mutant cultures. The mutant derivative yielded homogeneous while parental strain formed heterogeneous amplification patterns. Seven primers identified 42.9% polymorphism in the amplification products, indicating that these primers determined some genetic variability between the two strains. Thus RAPD was found to be an efficient technique to determine genetic variability in the mutant and wild organisms. Both wild and mutant strains were analyzed for their potential to produce galactosidases. Comparison of different carbon sources on enzyme yield revealed that wheat bran is significant (P < 0.01) effective producer and economical source followed by rice bran, rice polishing and lactose. The mutant was significantly better enzyme producer and could be considered for its prospective application in food, nutrition and health and that RAPD can be effectively used to differentiate mutant strain from the parental strain based on the RAPD patterns.

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References

  1. Wang CL, Li DF, Lu WQ, Wang YH, Lai CH (2004) Influence of cultivating conditions on the α-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation. Lett Appl Microbiol 39:369–375

    Article  CAS  PubMed  Google Scholar 

  2. Liu CQ, Chen QH, Tang B, Ruan H, He GQ (2007) Response surface methodology for optimizing the fermentation medium of alpha-galactosidase in solid state fermentation. Lett Appl Microbiol 45:206–212

    Article  CAS  PubMed  Google Scholar 

  3. Anisha GS, Sukumaran RK, Prema P (2008) Evaluation of alpha-galactosidase biosynthesis by Streptomyces griseoalbus in solid-state fermentation. Lett Appl Microbiol 46:338–345

    Article  CAS  PubMed  Google Scholar 

  4. Shankar SK, Mulimani VH (2007) Alpha-galactosidase production by Aspergillus oryzae in solid-state fermentation. Bioresour Technol 98:958–961

    Article  CAS  PubMed  Google Scholar 

  5. Oliveira O, Teixeira JA, Lima N (2007) Development of stable flocculent Saccharomyces cerevisiae strain for continuous Aspergillus niger β-galactosidase production. J Biosci Bioeng 103:212–218

    Article  Google Scholar 

  6. Yanai K, Nakane A, Hirayama M (2001) Molecular cloning and characterization of the fructo-oligosaccharide producing β-galactosidase gene from Aspergillus niger ATCC 20611. Biosci Biotechnol Biochem 4:466–473

    Google Scholar 

  7. Rajoka MI, Latif F, Khan S, Shahid R (2004) Kinetics of improved productivity of β-galactosidase by a cycloheximide-resistant mutant of Kluyveromyces marxianu. Biotechnol Lett 26:741–746

    Article  CAS  PubMed  Google Scholar 

  8. Viniegra-Gonzalez G, Favela-Torres E (2006) Why solid-state fermentation seems to be resistant to catabolite repression? Food Technol Biotechnol 44:397–406

    CAS  Google Scholar 

  9. Haq I, Khurshaid S, Ali S, Ashraf H, Qadeer MA, Rajoka MI (2001) Mutation of Aspergillus niger for hyper-production of citric acid from black strap molasses. World J Microbial Biotechnol 17:35–37

    Article  Google Scholar 

  10. Rajoka MI, Bashir A, Hussain MRA, Ghauri MT, Malik KA (1998) Mutagenesis of Cellulomonas biazotea for improved production of cellulases. Folia Microbiol 43:15–22

    Article  CAS  Google Scholar 

  11. Rincon AM, Codon AC, Castrejon F, Benitez T (2001) Improved properties of baker’s yeast mutant resistant to 2-deoxy-d-glucose. Appl Environ Microbiol 67:4279–4285

    Article  CAS  PubMed  Google Scholar 

  12. Liu Z, Yang X, Sun D, Song J, Chen G, Juba O, Yang Q (2010) Expressed sequence tags-based identification of genes in a biocontrol strain Trichoderma asperellum. Mol Biol Rep. doi:10.1007/s11033-010-0019-0 (available online)

  13. Ghally M, Ramos AM, Dokmetzian D, Lopez SE (2007) Genetic variability of Phytophthora sojae isolates from Argentina. Mycologia 99:877–883

    Article  Google Scholar 

  14. Ashokkumar B, Gunasekaran P (2002) β-Fructofuranosidase production by 2-deoxyglucose resistant mutants of Aspergillus niger in submerged and solid-state fermentation. Indian J Exp Biol 40:1032–1037

    CAS  PubMed  Google Scholar 

  15. Smith DC, Wood TM (1991) Isolation of mutants of Aspergillus awamori with enhanced production of extracellular xylanase and β-xylosidase. World J Microbiol Biotechnol 7:343–354

    Article  CAS  Google Scholar 

  16. Bradford MM (1976) A rapid and sensitive method for the quantification of micro-organism quantities of proteins utilizing the principles of protein dye-binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  17. Miller GL (1959) Use of dinitrosalicylic reagent for determination of reducing sugars. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  18. Shirlaw DWG (1969) A practical course in agriculture chemistry. Pergamon Press, New York

    Google Scholar 

  19. Pirt SJ (1975) Principles of cell cultivation. Blackwell Scientific, London. ISBN 0-63208150-3

  20. Zhang AL, Zhang TY, Luo JX, Fu CY, Qu Z, Yi GH, Su DX, Tu FZ, Pan YW (2009) Inducible expression of human angiostatin by AOXI promoter in Pichia pastoris using high-density cell culture. Mol Biol Rep 36(8):2265–2270

    Article  CAS  PubMed  Google Scholar 

  21. Afzal S, Saleem M, Yasmin R, Naz M, Imran M (2010) Pre and post cloning characterization of a beta-1,4-endoglucanase from Bacillus sp. Mol Biol Rep 37(4):1717–1723

    Article  CAS  PubMed  Google Scholar 

  22. Yoshimitsu M, Higuchi K, Fan X, Takao S, Medin JA, Tei C, Takenaka T (2010) Sequencing and characterization of the porcine alpha-galactosidase A gene: towards the generation of a porcine model for Fabry disease. Mol Biol Rep. doi:10.1007/s11033-010-9985-S (available online)

  23. Hrmova M, Petrakova E, Biely P (1991) Induction of cellulose- and xylan-degrading enzyme systems in Aspergillus terreus by homo- and hetero-disaccharides composed of glucose and xylose. J Gen Microbiol 137:541–547

    CAS  PubMed  Google Scholar 

  24. Gao J, Weng H, Zhee D, Yuan M, Guan F, Xi Y (2008) Production and characterization of cellulolytic enzymes from the thermoacidophilic fungal Aspergillus terreus M11 under solid-state cultivation of corn Stover. Bioresour Technol 99:7623–7629

    Article  CAS  PubMed  Google Scholar 

  25. Ademark P, Larsson M, Tjerneld F, Stålbrand H (2003) Multiple α-galactosidases from Aspergillus niger: purification, characterization and substrate specificity. Enzym Microb Technol 29:441–448

    Article  Google Scholar 

  26. Kuhad RC, Kapoor M, Rustagi RE (2004) Enhanced production of an alkaline pectinase from Streptomyces sp. RCK-SC by whole cell immobilization and solid-state cultivation. World J Microbiol Biotechnol 20:35–37

    Article  Google Scholar 

  27. Kondoh K, Morisaki K, Kim WO, Park GG, Kaneko S, Kobayashi H (2005) Cloning and expression of the gene encoding Streptomyces coelicolor A3 (2) α-galactosidase belonging to family 36. Biotechnol Lett 27:641–647

    Article  CAS  PubMed  Google Scholar 

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Correspondence to M. E. Babar.

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Siddique Awan, M., Tabbasam, N., Ayub, N. et al. Gamma radiation induced mutagenesis in Aspergillus niger to enhance its microbial fermentation activity for industrial enzyme production. Mol Biol Rep 38, 1367–1374 (2011). https://doi.org/10.1007/s11033-010-0239-3

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  • DOI: https://doi.org/10.1007/s11033-010-0239-3

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