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Brazilian Journal of Microbiology

, Volume 50, Issue 1, pp 53–65 | Cite as

A new strain of Aspergillus tubingensis for high-activity pectinase production

  • Danmei Huang
  • Yuyang Song
  • Yanlin Liu
  • Yi QinEmail author
Biotechnology and Industrial Microbiology - Research Paper
  • 10 Downloads

Abstract

Pectinase is a general term for a class of enzymes that decompose pectin. To obtain a fungal strain with high-activity pectinase of potential commercial importance, we screened microorganisms from the soil of vineyards, performed mutation breeding by ultraviolet (UV) and nitrosoguanidine (NTG) mutagenesis, and performed comparisons to commercially available pectinases. We found that the derived pectinase-producing strain Rn14-88A had the highest pectinase activity of 8363.215 U/mL, and identified it using internal transcribed spacer sequence analysis as Aspergillus tubingensis. Rn14-88A was the original strain for UV mutagenesis, from which mutant strain R-7-2-4 had the highest pectinase enzyme activity (9198.68 U/mL), which was a 9.99% increase compared to that of Rn14-88A. Following NTG mutagenesis of R-7-2-4, mutant strain Y1-3-2-6 had a pectinase enzyme activity of 9843.34 U/mL, which reflects a 6.36% increase compared to the pectinase activity of R-7-2-4. Subsequently, another round of NTG mutagenesis was performed on Y1-3-2-6, and the mutagenic strain Y2-6-3-4 exhibited an improved enzyme activity of 21,864.34 U/mL, which was 161.44% higher than that of Rn14-88A. Through liquid fermentation experiments of A. tubingensis Y2-6-3-4, it was determined that pectinase activity was the highest at a fermentation time of 20 h. Therefore, we conclude that A. tubingensis Y2-6-3-4 has potential for use in commercial production.

Keywords

Screening Pectinase Aspergillus tubingensis Ultraviolet mutagenesis (UV) Nitrosoguanidine (NTG) mutagenesis 

Notes

Funding information

The study was funded by the National Key R&D Program of China (2016YFD0400504-01), Viticulture Experiment Station scientific and technological transformative project of Northwest A&F University (TGZX2017-23), Scientific and Technological Transformative Special Project of Shanxi Province (201604D132034), and China Agriculture Research System (grant no. CARS-29-jg-3).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. 1.
    Sittidilokratna C, Suthirawut S, Chitradon L, Punsuvon V, Basu AK (2007) Screening of pectinase producing bacteria and their efficiency in biopulping of paper mulberry bark. Sci Asia 33:1513–1874CrossRefGoogle Scholar
  2. 2.
    Samaranayake CP, Sastry SK (2016) Effects of controlled-frequency moderate electric fields on pectin methylesterase and polygalacturonase activities in tomato homogenate. Food Chem 199:265–272CrossRefGoogle Scholar
  3. 3.
    Almeida DPF, Huber DJ (2007) Polygalacturonase-mediated dissolution and depolymerization of pectins in solutions mimicking the pH and mineral composition of tomato fruit apoplast. Plant Sci 172(6):1087–1094CrossRefGoogle Scholar
  4. 4.
    Pérezfuentes C, Cristina RM, Eyzaguirre J (2014) Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization. Fungal Biol-UK 118(5–6):507–515CrossRefGoogle Scholar
  5. 5.
    Abusarra AF, Abugoukh AA (2016) Changes in pectinesterase, polygalacturonase and cellulase activity during mango fruit ripening. J Hortic Sci Biotechnol 67(4):561–568CrossRefGoogle Scholar
  6. 6.
    Camassola M, Bittencourt LRD, Shenem NT, Andreaus J, Dillon AJP (2004) Characterization of the cellulase complex of Penicillium echinulatum. Biocatal Biotransfor 22(5–6):391–396CrossRefGoogle Scholar
  7. 7.
    Quintero RN, Cordoba CY, Stashenko EE, Fuentes JL (2017) Antigenotoxic effect against ultraviolet radiation induced DNA damage of the essential oils from lippia species. Photechem Photobiol 93(4):1063–1072CrossRefGoogle Scholar
  8. 8.
    Wang S, Zhang L, Yang GP, Han JC, Thomsen L, Pan KH (2016) Breeding 3 elite strains of Nannochloropsis oceanica, by nitrosoguanidine mutagenesis and robust screening. Algal Res 19:104–108CrossRefGoogle Scholar
  9. 9.
    Yin LB, Zhang CF, Xia QL, Yang Y, Xiao K (2016) Enhancement of pectinase production by ultraviolet irradiation and diethyl sulfate mutagenesis of a Fusarium oxysporum isolate. Genet Mol Res 15(3):86–89Google Scholar
  10. 10.
    Harper M, Lee CJ (2012) Genome-wide analysis of mutagenesis bias and context sensitivity of N-methyl-N’-nitro-N-nitrosoguanidine (NTG). Mutat Res 731(1–2):64–67CrossRefGoogle Scholar
  11. 11.
    Jay JM, Loessner MJ, Golden DA (2000) Modern food microbiology. Bioscience 101(3):167–176Google Scholar
  12. 12.
    Patil SR, Dayanand A (2006) Exploration of regional agrowastes for the production of pectinase by Aspergillus niger. Food Technol Biotechnol 44(2):289–292Google Scholar
  13. 13.
    Periyasamy K, Santhalembi L, Mortha G, Aurousseau M, Guillet A, Dallerac D, Sivanesan S (2017) Production, partial purification and characterization of enzyme cocktail from Trichoderma citrinoviride, AUKAR04 through solid-state fermentation. Arab J Sci Eng 42(1):53–63CrossRefGoogle Scholar
  14. 14.
    Reetha S, Selvakumar G, Thamizhiniyan P, Ravimycin T, Bhuvaneswari G (2014) Screening of cellulase and pectinase by using Pseudomonas fluorescence and Bacillus subtilis. IEEE 13:75–80Google Scholar
  15. 15.
    Yu P, Zhang Y, Gu D (2007) Production optimization of a heat-tolerant alkaline pectinase from Bacillus subtilis ZGl14 and its purification and characterization. J Appl Phys 101(6):21–24Google Scholar
  16. 16.
    Shelomi M, Danchin EGJ, Heckel D, Wipfler B (2016) Horizontal gene transfer of pectinases from bacteria preceded the diversification of stick and leaf insects. Sci Rep 6:26388–26397CrossRefGoogle Scholar
  17. 17.
    Owen AB (2007) The dimension distribution and quadrature test functions. Stat Sin 13(13):1–17Google Scholar
  18. 18.
    Poletto P, Polidoro TA, Zeni M, Silveira MMD (2017) Evaluation of the operating conditions for the solid-state production of pectinases by Aspergillus niger, in a bench-scale, intermittently agitated rotating drum bioreactor. LWT-Food Sci Technol 79:92–101CrossRefGoogle Scholar
  19. 19.
    Finkler ATJ, Biz A, Pitol LO, Medina BS (2017) Intermittent agitation contributes to uniformity across the bed during pectinase production by Aspergillus niger, grown in solid-state fermentation in a pilot-scale packed-bed bioreactor. Biochem Eng J 121:1–12CrossRefGoogle Scholar
  20. 20.
    Lee L, Arul J, Lencki R, Castaigne F (2010) A review on modified atmosphere packaging and preservation of fresh fruits and vegetables: physiological basis and practical aspects-part I. Packag Technol Sci 8(6):315–331CrossRefGoogle Scholar
  21. 21.
    Zhang W, Wang M, Huang Y, Chea SKP, Zheng Z (2011) New and highly efficient methodology for screening high-yield strains of cytotoxic deacetylmycoepoxydiene (DAM). Lett Appl Microbiol 52(5):441–447CrossRefGoogle Scholar
  22. 22.
    Xavier-Santos S, Carvalho CC, Bonfá M, Silva R, Capelari M (2004) Screening for pectinolytic activity of wood-rotting basidiomycetes and characterization of the enzymes. Folia Microbiol 49(1):46–52CrossRefGoogle Scholar
  23. 23.
    Ruiz HA, Rodríguez-Jasso RM, Rodríguez R, Contreras-Esquivel JC (2012) Pectinase production from lemon peel pomace as support and carbon source in solid-state fermentation column-tray bioreactor. Biochem Eng J 65(5):90–95CrossRefGoogle Scholar
  24. 24.
    Sukrong S, Zhu S, Ruangrungsi N, Phadungcharoen T, Palanuvej C, Komatsu K (2007) Molecular analysis of the genus Mitragyna existing in Thailand based on rDNA ITS sequences and its application to identify a narcotic species: Mitragyna speciosa. Biol Pharm Bull 30(7):1284–1288CrossRefGoogle Scholar
  25. 25.
    Bibi N, Ali S, Tabassum R (2016) Statistical optimization of pectinase biosynthesis from orange peel by Bacillus licheniformis, using submerged fermentation. Waste Biomass Valoriz 7(3):467–481CrossRefGoogle Scholar
  26. 26.
    Khadija A, Ahmad Z, Ghulam R, Sajjad MM (2016) Isolation and identification by 16S rRNA sequence analysis of plant growth-promoting azospirilla from the rhizosphere of wheat. Braz Soc Microbiol 47(3):542–550CrossRefGoogle Scholar
  27. 27.
    Ben-Arie R, Sonego L (1980) Pectolytic enzyme activity involved in woolly breakdown of stored peaches. Phytochemistry 19(12):2553–2555CrossRefGoogle Scholar
  28. 28.
    Lewis KC, Selzer T, Shahar C, Udi Y, Tworowski D, Irit S (2008) Inhibition of pectin methyl esterase activity by green tea catechins. Phytochemistry 69(14):2586–2592CrossRefGoogle Scholar
  29. 29.
    Lott JA, Stephan VA, Jr PK (1983) Evaluation of the Coomassie Brilliant Blue G-250 method for urinary protein. Clin Chem 29(11):1946–1950Google Scholar
  30. 30.
    Kittur FS, Kumar ABV, Gowda LR, Tharanathan RN (2003) Chitosanolysis by a pectinase isozyme of Aspergillus niger-a non-specific activity. Carbohydr Polym 53(2):191–196CrossRefGoogle Scholar
  31. 31.
    Sharma S, Mandhan RP, Sharma J (2012) Utilization of agro-industrial residues for pectinase production by the novel strain Pseudozyma, sp. SPJ under solid state cultivation. Ann Microbiol 62(1):169–176CrossRefGoogle Scholar
  32. 32.
    Geetha M, Saranraj P, Mahalakshm S, Reetha D (2012) Screening of pectinase producing bacteria and fungi for its pectinolytic activity using fruit wastes. Int J Biochem Biotech Sci 1:30–42Google Scholar
  33. 33.
    Zhang CLUV (2009) NTG mutagenesis for breeding of a high trehalose-producing strain of Saccharomyces cerevisiae. Food Sci 30(21):188–191Google Scholar
  34. 34.
    Sherrer SM, Taggart DJ, Pack LR, Malik CK, Basu AK, Suo Z (2012) Quantitative analysis of the mutagenic potential of 1-aminopyrene-DNA adduct bypass catalyzed by Y-family DNA polymerases. Mutat Res 737(1–2):25–33CrossRefGoogle Scholar

Copyright information

© Sociedade Brasileira de Microbiologia 2019

Authors and Affiliations

  • Danmei Huang
    • 1
  • Yuyang Song
    • 1
    • 2
  • Yanlin Liu
    • 1
    • 2
  • Yi Qin
    • 1
    • 2
    Email author
  1. 1.College of EnologyNorthwest A&F UniversityYanglingChina
  2. 2.Shaanxi Engineering Research Center for Viti-VinicultureYanglingChina

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