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Screening of pectinase-producing bacteria from farmlands and optimization of enzyme production from selected strain by RSM

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Abstract

Pectinolytic enzymes that catalyze the breakdown of substrates containing pectin are widespread. Pectinases have potential applications in various industries, including food, animal feed, textile, paper, and fuel. In this study, one hundred bacterial isolates were collected from Marand city farmlands (Azarbaijan-E-Sharqi, Iran) and screened by MP medium on the base of pectinase activity considering the significance of pectinases. The results depicted that three isolates showed the most pectinase activity (more massive halo). The biochemical and molecular test results showed that the three screened bacteria were Enterobacter and named Enterobacter sp. MF41, Enterobacter sp. MF84, and Enterobacter sp. MF90. Enterobacter sp. MF84 had the largest halo, so this strain was selected for the study of its produced pectinase. The results exhibited that the produced enzyme has optimum temperature and pH for activity at 30 °C and in 9, respectively. Finally, the enzyme production by Enterobacter sp. MF84 is optimized using response surface methodology (RSM) considering four factors (NH4Cl, K2HPO4, pectin, and incubation time) as variables. The results showed that the optimization procedure increased the enzyme production up to 12 times (from 1.16 to 14.16 U/mg). The Pareto analysis revealed that ammonium chloride has a significant role in decreasing the enzyme production, probably by inducing the nitrification pathway enzymes in the presence of organic nitrogen in Enterobacter sp. MF84.

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Abbreviations

ANOVA:

Analysis of variance

CTAB:

Cetyltrimethylammonium Bromide

LOF:

Lack of fit

RSM:

Response surface methodology

References

  • Afzal I, Shinwari ZK, Iqrar I (2015) Selective isolation and characterization of agriculturally beneficial endophytic bacteria from wild hemp using canola. Pak J Bot 47:1999–2008

    CAS  Google Scholar 

  • Ahmed I, Zia MA, Hussain MA, Akram Z, Naveed MT, Nowrouzi A (2016) Bioprocessing of citrus waste peel for induced pectinase production by Aspergillus niger; its purification and characterization. J Radiat Res Appl Sci 9:148–154

    CAS  Google Scholar 

  • Anuradha K, Padma PN, Venkateshwar S, Reddy G (2016) Mango juice clarification with polygalacturonase produced by Aspergillus awamori MTCC 9166-optimization of conditions. Int Food Res J 23:147

    CAS  Google Scholar 

  • Basu S, Ghosh A, Bera A, Saha MN, Chattopadhyay D, Chakrabarti K (2008) Thermodynamic characterization of a highly thermoactive extracellular pectate lyase from a new isolate Bacillus pumilus DKS1. Bioresour Technol 99:8088–8094

    CAS  PubMed  Google Scholar 

  • Beulah D, Sunitha E, Srilakshmi T (2015) Production, purification and assay of pectinase enzyme from Aspergillus niger. Helix 2:673–677

    Google Scholar 

  • Bibi N, Ali S, Tabassum R (2016) Statistical optimization of pectinase biosynthesis from orange peel by Bacillus licheniformis using submerged fermentation. Waste Biomass Valori 7:467–481

    CAS  Google Scholar 

  • Blanco P, Sieiro C, Villa TG (1999) Production of pectic enzymes in yeasts. FEMS Microbiol Lett 175:1–9

    CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    CAS  Google Scholar 

  • Brühlmann F, Kim KS, Zimmerman W, Fiechter A (1994) Pectinolytic enzymes from actinomycetes for the degumming of ramie bast fibers. Appl Environ Microbiol 60:2107–2112

    PubMed  PubMed Central  Google Scholar 

  • Chandra Reddy MP, Saritha K (2016) Effects of the culture media optimization on pectinase production by Enterobacter sp. PSTB-1. 3 Biotech 6:207

    Google Scholar 

  • Darah I, Nisha M, Lim S (2013) Enhancement of polygalacturonase production from Enterobacter aerogenes NBO2 by submerged fermentation. Adv Stud Biol 5:173–189

    Google Scholar 

  • Handa S, Sharma N, Pathania S (2016) Multiple parameter optimization for maximization of pectinase production by Rhizopus sp. C4 under solid state fermentation. Fermentation 2:10

    Google Scholar 

  • Hoondal G, Tiwari R, Tewari R, Dahiya N, Beg Q (2002) Microbial alkaline pectinases and their industrial applications: a review. Appl Microbiol Biotechnol 59:409–418

    CAS  PubMed  Google Scholar 

  • Huang LK, Mahoney R (1999) Purification and characterization of an endo-polygalacturonase from Verticillium albo-atrum. J Appl Microbiol 86:145–156

    CAS  Google Scholar 

  • Jayani RS, Saxena S, Gupta R (2005) Microbial pectinolytic enzymes: a review. Process Biochem 40:2931–2944

    CAS  Google Scholar 

  • Joshi M, Nerurkar M, Adivarekar R (2015) Characterization, kinetic, and thermodynamic studies of marine pectinase from Bacillus subtilis. Prep Biochem Biotechnol 45:205–220

    CAS  PubMed  Google Scholar 

  • Kashyap D, Vohra P, Chopra S, Tewari R (2001) Applications of pectinases in the commercial sector: a review. Bioresour Technol 77:215–227

    CAS  PubMed  Google Scholar 

  • Kaur SJ, Gupta VK (2017) Production of pectinolytic enzymes pectinase and pectin lyase by Bacillus subtilis SAV-21 in solid state fermentation. Ann Microbiol 67:333–342

    CAS  Google Scholar 

  • Khataee AR, Zarei M, Asl SK (2010) Photocatalytic treatment of a dye solution using immobilized TiO2 nanoparticles combined with photoelectro-Fenton process: optimization of operational parameters. J Electroanal Chem 648:143–150

    CAS  Google Scholar 

  • Khatri BP, Bhattarai T, Shrestha S, Maharjan J (2015) Alkaline thermostable pectinase enzyme from Aspergillus niger strain MCAS2 isolated from Manaslu Conservation Area, Gorkha, Nepal. Springerplus 4:488

    PubMed  PubMed Central  Google Scholar 

  • Klug-Santner BG, Schnitzhofer W, Vrsanská M, Weber J, Agrawal PB, Nierstrasz VA, Guebitz GM (2006) Purification and characterization of a new bioscouring pectate lyase from Bacillus pumilus BK2. J Biotechnol 121:390–401

    CAS  PubMed  Google Scholar 

  • Kohli P, Gupta R (2015) Alkaline pectinases: a review. Biocatal Agric Biotechnol 4:279–285

    Google Scholar 

  • Lara-Márquez A, Zavala-Páramo MG, López-Romero E, Camacho HC (2011) Biotechnological potential of pectinolytic complexes of fungi. Biotechnol Lett 33:859–868

    PubMed  Google Scholar 

  • Li P-J et al (2015) Optimizing production of pectinase from orange peel by Penicillium oxalicum PJ02 using response surface methodology. Waste Biomass Valoriz 6:13–22

    Google Scholar 

  • Mahdinia E, Demirci A, Berenjian A (2019) Effects of medium components in a glycerol-based medium on vitamin K (menaquinone-7) production by Bacillus subtilis natto in biofilm reactors. Bioprocess Biosyst Eng 42:223–232

    CAS  PubMed  Google Scholar 

  • Maleki MH, Ghanbary MAT, Ranjbar G, Asgharzadeh A, Lotfi A (2017) Screening of some Zygomycetes strains for pectinase activity. J Microbiol Biotechnol Res 1:1–7

    Google Scholar 

  • Martos MA, Zubreski ER, Garro OA, Hours RA (2013) Production of Pectinolytic enzymes by the yeast Wickerhanomyces anomalus isolated from citrus fruits peels. Biotechnol Res Int 2013: 435154 

  • Mathew A, Eldo AN, Molly A (2008) Optimization of culture conditions for the production of thermostable polygalacturonase by Penicillium SPC-F 20. J Ind Microbiol Biotechnol 35:1001–1005

    CAS  PubMed  Google Scholar 

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

    CAS  Google Scholar 

  • Nawaz MA, Bhattarai RR (2015) Isolation and identification of Yersinia and Pseudomonas sp. from Australian milk and salad using 16s rDNA. In PeerJ PrePrints 3:e820v1

    Google Scholar 

  • Padhi SK, Tripathy S, Mohanty S, Maiti NK (2017) Aerobic and heterotrophic nitrogen removal by Enterobacter cloacae CF-S27 with efficient utilization of hydroxylamine. Bioresour Technol 232:285–296

    CAS  PubMed  Google Scholar 

  • Patil SR, Dayanand A (2006) Optimization of process for the production of fungal pectinases from deseeded sunflower head in submerged and solid-state conditions. Bioresour Technol 97:2340–2344

    CAS  PubMed  Google Scholar 

  • Pedrolli DB, Monteiro AC, Gomes E, Carmona EC (2009) Pectin and pectinases: production, characterization and industrial application of microbial pectinolytic enzymes. Open Biotechnol J 3:9–18

    CAS  Google Scholar 

  • Purnachandra Reddy M, Saritha K (2014) Studies on pectinase production by Enterobacter sp. using mango fruit processing industrial waste as whole and sole carbon source. Curr Trends Biotechnol Pharm 8:378–386

    Google Scholar 

  • Rehman HU, Aman A, Nawaz MA, Qader SAU (2015) Characterization of pectin degrading polygalacturonase produced by Bacillus licheniformis KIBGE-IB21. Food Hydrocoll 43:819–824

    Google Scholar 

  • Schaad NW, Jones JB, Chun W (2001) Laboratory guide for the identification of plant pathogenic bacteria. 3rd edn. American Phytopathological Society (APS Press). Saint Paul, Minnesota, USA. 

  • Shah KP, Chandok KH, Rathore P, Sharma MV, Yadav M, Nayarisseri SA (2013) Screening, isolation and identification of polygalacturonase producing Bacillus tequilensis strain EMBS083 using 16S rRNA gene sequencing. Eur J Biol Sci 5:09–13

    Google Scholar 

  • Sharma A, Husain I (2015) Optimization of medium components for extracellular glutaminase free asparaginase from Enterobacter cloacae. Int J Curr Microbiol App Sci 4:296–309

    CAS  Google Scholar 

  • Sharma N, Rathore M, Sharma M (2013) Microbial pectinase: sources, characterization and applications. Rev Environ Sci Biotechnol 12:45–60

    CAS  Google Scholar 

  • Singh S, Mandal SK (2012) Optimization of processing parameters for production of pectinolytic enzymes from fermented pineapple residue of mixed Aspergillus species. Jordan J Biol Sci 147:1–7

    Google Scholar 

  • Sohail M, Latif Z (2016) Phylogenetic analysis of polygalacturonase-producing Bacillus and Pseudomonas isolated from plant waste material. Jundishapur J Microbiol 9:e28594

    PubMed  PubMed Central  Google Scholar 

  • Thangaratham T, Manimegalai G (2014) Optimization and production of pectinase using agro waste by solid state and submerged fermentation. Int J Curr Microbiol App Sci 3:357–365

    CAS  Google Scholar 

  • Uzuner S, Cekmecelioglu D (2015) Enhanced pectinase production by optimizing fermentation conditions of Bacillus subtilis growing on hazelnut shell hydrolyzate. J Mol Catal B Enzym 113:62–67

    CAS  Google Scholar 

  • Venzon SS, Canteri MH, Granato D, Junior BD, Maciel GM, Stafussa AP, Haminiuk CW (2015) Physicochemical properties of modified citrus pectins extracted from orange pomace. J Food Sci Technol 52:4102–4112

    CAS  PubMed  Google Scholar 

  • Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey, FA, Schleifer, K-H, Whitman, WB (2011) Bergey's manual of systematic bacteriology: Volume 3: The Firmicutes. Springer Science & Business Media, Heidelberg, Germany. 

  • Yadav S, Yadav P, Yadav D, Yadav K (2009) Pectin lyase: a review. Process Biochem 44:1–10

    Google Scholar 

  • Yuan P, Meng K, Shi P, Luo H, Huang H, Tu T, Yang P, Yao B (2012) An alkaline-active and alkali-stable pectate lyase from Streptomyces sp. S27 with potential in textile industry. J Ind Microbiol Biotechnol 39:909–915

    CAS  PubMed  Google Scholar 

  • Zhou M, Wu J, Wang T, Gao L, Yin H, Lü X (2017) The purification and characterization of a novel alkali-stable pectate lyase produced by Bacillus subtilis PB1. World J Microbiol Biotechnol 33:190

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors express their gratitude to the research council of Azarbaijan Shahid Madani University for the financial support during the course of this project.

Funding

This work was supported by the research council of Azarbaijan Shahid Madani University (Tabriz, Iran).

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Authors and Affiliations

  1. Department of Cellular and Molecular Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran

    Rezvan Abdollahzadeh, Mohammad Pazhang & Saeed Najavand

  2. Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran

    Vahid Fallahzadeh-Mamaghani

  3. Applied Chemistry Research Laboratory, Department of Chemistry, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran

    Ali Reza Amani-Ghadim

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  1. Rezvan Abdollahzadeh
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  2. Mohammad Pazhang
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  4. Vahid Fallahzadeh-Mamaghani
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  5. Ali Reza Amani-Ghadim
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Correspondence to Mohammad Pazhang.

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Abdollahzadeh, R., Pazhang, M., Najavand, S. et al. Screening of pectinase-producing bacteria from farmlands and optimization of enzyme production from selected strain by RSM. Folia Microbiol 65, 705–719 (2020). https://doi.org/10.1007/s12223-020-00776-7

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  • DOI: https://doi.org/10.1007/s12223-020-00776-7

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