Isolation and screening of pectinase producing Aspergillus niger and use of whey to optimize enzyme production
Subject Areas : Microbiology
Mahsa pouya
1
,
Shokoofeh Ghazi
2
,
امیر توکمه چی
3
1 - 1M.Sc of Industrial Microbiology, Department of Microbiology, Faculty of New Science, Tehran Medical sciences, Islamic Azad university, Tehran, Iran
2 - Department of Microbiology, Faculty of Basic Science, Tehran Medical Branch, Islamic Azad University, Tehran, Iran
3 - گروه میکروبیولوژی، دانشکده دامپزشکی، دانشگاه ارومیه
Keywords: Aspergillus niger, Pectinase enzyme, DNS method, Whey,
Abstract :
Abstract Aims and Background: Pectinase that can break down pectin in plant cell walls is one of the most important industrial enzymes in the world, which can be isolated from a wide range of microorganisms such as bacteria and fungi. Therefore, considering the importance and applications of this enzyme in food processing, agriculture, the purpose of this research is isolating and screening pectinase-producing fungi from some of rotten fruits and vegetables, as well as optimizing the enzyme production condition. Materials and methods: Isolation and identification of pectinase producing fungi from rotten fruits (peach, plum and apple) and vegetables (onion) were done. Screening the most capable fungal strains producing pectinase was used by cultivation on specific Pectin Agar medium, staining with lactophenol cotton blue and slide culture method. To identify the isolated fungi more accurately, the molecular technique such as 18S rRNA sequencing was used. Optimizing the production rate of pectinase enzyme carried out using pectin substrate accompanied by different Lactic Whey and Permeate whey under different temperatures and pH levels, and the activity of enzyme was measured by the standard DNS method. Results: one Aspergillus niger fungus producing pectinase enzyme was isolated from onion. Results for optimization conditions shows that the identified strain, shows an enzyme activity equal to 103/018 (IU/ml) in the presence of permeate whey and pectin substrate, under incubation at 25 C° ,140 rpm, and pH 6.5. While, pectinase production with only pure pectin substrate was lower equal to 92.717 (IU/ml) when incubation at 25 C° , same rpm and pH 7. Conclusions: Potent native fungal strains isolated from our country are suitable candidates for the production of important industrial enzymes such as Pectinase. Also using cost effective substrates like Whey can be replaced by expensive pure substrates in production industry for the important enzymes production process.
1. Sandri IG, Lorenzoni CM, Fontana RC, da Silveira MM. Use of pectinases produced by a new strain of Aspergillus niger for the enzymatic treatment of apple and blueberry juice. LWT-Food Science and Technology. 2013 May 1;51(2):469-75.
2. Songol, A., Behbahani, M. Isolation and optimization of pectinase enzyme production one of useful industrial enzyme in Aspergillus niger, Rhizopus oryzae, Penicilium chrysogenum. Biological Journal of Microorganism, 2016; 5(17): 121-140. doi: 10.22108/bjm.2016.20374.
3. Singh R, Kumar M, Mittal A, Mehta PK. Microbial enzymes: industrial progress in 21st century. 3 Biotech. 2016 Dec;6(2):1-5.
4. KC S, Upadhyaya J, Joshi DR, Lekhak B, Kumar Chaudhary D, Raj Pant B, Raj Bajgai T, Dhital R, Khanal S, Koirala N, Raghavan V. Production, characterization, and industrial application of pectinase enzyme isolated from fungal strains. Fermentation. 2020 Jun 9;6(2):59.
5. Jayani RS, Saxena S, Gupta R. Microbial pectinolytic enzymes: a review. Process Biochemistry. 2005 Sep 1;40(9):2931-44.
6. Borszcz V, Boscato TR, Cenci K, Zeni J, Cansian RL, Backes GT, Valduga E. Extraction conditions evaluation of pectin methylesterase produced by solid state culture of Aspergillus niger. Czech Journal of Food Sciences. 2019 Jan 7;36(6):476-9.
7. Rosana Y, Matsuzawa T, Gonoi T, Karuniawati A. Modified slide culture method for faster and easier identification of dermatophytes. Microbiology indonesia. 2014 Sep 5;8(3):7-.
8. Sutradhar P, Saha I, Chakravarty A. Evaluation of pomegranate peel as a substrate for citric acid production by Aspergillus niger. International Journal of Current Pharmaceutical Research. 2020 May 15:61-5.
9. Karazhyan R., Habibi Najafi M.H., Yavarmanesh M., Edalatian Dovom M.R., Pourianfar H.R. Comparison of optimized DNA extraction from mold Aspergillus, Penicillium and Rhizopus of tomato paste. Iranian Journal of Food Sience and Technology. 2017 [cited 2022September05];14(67):1-9.
10. Susca A, Stea G, Mulé G, Perrone G. Polymerase chain reaction (PCR) identification of Aspergillus niger and Aspergillus tubingensis based on the calmodulin gene. Food Additives and Contaminants. 2007 Oct 1;24(10):1154-60.
11. Oumer OJ, Abate D. Screening and molecular identification of pectinase producing microbes from coffee pulp. BioMed research international. 2018 Apr 3;2018.
12. Chowdhury TI, Jubayer MF, Uddin MB, Aziz MG. Production and characterization of pectinase enzyme from rhizopus oryzae. Potravinarstvo. 2017.
13. El-Holi MA, Al-Delaimy S. Citric acid production from whey with sugars and additives by Aspergillus niger. African Journal of Biotechnology. 2003;2(10):356-9.
14. Preeti S, Abhishek T, Deeja K, Suresh S. Isolation, screening and optimization of novel pectinase producing fungal strain for fruit juice clarification and extraction. World Journal of Pharmaceutical Research. 2015;4(6):2114-26.
15. Beg QK, Bhushan B, Kapoor M, Hoondal GS. Production and characterization of thermostable xylanase and pectinase from Streptomyces sp. QG-11-3. Journal of Industrial Microbiology and Biotechnology. 2000 Jun;24(6):396-402.
16. Mahesh NA, Vivek RA, Arunkumar MA, Balakumar SR. Statistical designing of enriched pectin extract medium for the enhanced production of pectinase by Aspergillus niger. Inter. J. Pharm. Pharm. Sci. 2014 Feb;6(SUPPL 2):666-72.
17. Gophanea SR, Khobragadea CN, Jayebhayea SG. Extracellular pectinase activity from Bacillus Cereus GC subgroup a: isolation, production, optimization and partial characterisation. Journal of Microbiology, Biotechnology and Food Sciences. 2021 Jan 6;2021:767-72.
18. Palaniyappan M, Vijayagopal V, Viswanathan R, Viruthagiri T. Statistical optimization of substrate, carbon and nitrogen source by response surface methodology for pectinase production using Aspergillus fumigatus MTCC 870 in submerged fermentation. African Journal of Biotechnology. 2009;8(22).
19. Kuvvet C, Uzuner S, Cekmecelioglu D. Improvement of pectinase production by co-culture of Bacillus spp. using apple pomace as a carbon source. Waste and Biomass Valorization. 2019 May;10(5):1241-9.
