Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran
Introduction:Pectinase or pectinolytic enzymes are complex enzymes which include pectin methyl esterase (EC.3.1.1.11), pectin lyase (EC.4.2.2.2) and polygalacturonase (EC.3.3.1.15) which degrade pectin in the cell wall of plant cells. These enzymes have many industrial applications that some of them...
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| Format: | Article |
| Language: | English |
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University of Isfahan
2018-06-01
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| Series: | Biological Journal of Microorganism |
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| Online Access: | http://bjm.ui.ac.ir/article_22300_0e7a5768e40e85fd9e36361895849738.pdf |
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| author | Zohre Nasrollahzadeh Eniseh Saleh-Ghamari Mohammad Tahmaseb Mohammad Ali Amoozegar |
| author_facet | Zohre Nasrollahzadeh Eniseh Saleh-Ghamari Mohammad Tahmaseb Mohammad Ali Amoozegar |
| author_sort | Zohre Nasrollahzadeh |
| collection | DOAJ |
| description | Introduction:Pectinase or pectinolytic enzymes are complex enzymes which include pectin methyl esterase (EC.3.1.1.11), pectin lyase (EC.4.2.2.2) and polygalacturonase (EC.3.3.1.15) which degrade pectin in the cell wall of plant cells. These enzymes have many industrial applications that some of them are active in extreme condition regarding to temperature, pH and salt concentration. In this study, the bacteria with an ability to produce pectinase enzyme in salty condition were identified and the corresponding gene was analyzed.
Materials and methods: Strains from Urmia, Inche boron and Gomeshan Ponds were inoculated in the media containing pectin precursors. By analyzing the clear zones around the colonies based on I2/KI indicator, the positive strains were selected. Quantification of enzyme activity on all three types of pectinase was carried out by spectrophotometry. In order to molecularly screen the bacterium contained pectinase gene, the bacterium genome was amplified using appropriate primers.
Results: Seventeen positive strains for pectinase (10 from Gomeshan Lake, 6 from Inche Boron Lake and 1 from where Urmia Lake) were identified among 130 studied samples. According to size of clear zone of enzyme activity in the qualitative test, activity level of all three pectinase enzymes in R2S25 strain of Inche Boron Lake was measured and the growth curve was obtained. Molecular study showed that all strains contain desired gene segment.
Discussion and conclusion: Quantitative evaluation showed that production and activity of pectinase enzyme in R2S25 strain increased simultaneously with increasing the growth of selected strain in logarithmic phase. Molecular study also showed that the genuses of Martelella, Aeromocrobium, Planococcus, Marinobacter, Virgibacillus, Kocuria and Micrococcus contain the pectinase gene. |
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| id | doaj.art-aba74c007c13447291400ede15236f8e |
| institution | Directory Open Access Journal |
| issn | 2322-5173 2322-5181 |
| language | English |
| last_indexed | 2024-12-11T14:16:49Z |
| publishDate | 2018-06-01 |
| publisher | University of Isfahan |
| record_format | Article |
| series | Biological Journal of Microorganism |
| spelling | doaj.art-aba74c007c13447291400ede15236f8e2022-12-22T01:03:06ZengUniversity of IsfahanBiological Journal of Microorganism2322-51732322-51812018-06-0172611512210.22108/bjm.2017.77717.022300Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, IranZohre Nasrollahzadeh0Eniseh Saleh-Ghamari1Mohammad Tahmaseb2Mohammad Ali Amoozegar3M.Sc. Student of Genetics, Department of Cell and Molecular Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, IranAssistant Professor of Microbiology, Department of Cell and Molecular Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, IranAssistant Professor of Genetics, Department of Cell and Molecular Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, IranAssociate Professor of Microbiology, Department of Microbiology, Faculty of science, Tehran University, Tehran, IranIntroduction:Pectinase or pectinolytic enzymes are complex enzymes which include pectin methyl esterase (EC.3.1.1.11), pectin lyase (EC.4.2.2.2) and polygalacturonase (EC.3.3.1.15) which degrade pectin in the cell wall of plant cells. These enzymes have many industrial applications that some of them are active in extreme condition regarding to temperature, pH and salt concentration. In this study, the bacteria with an ability to produce pectinase enzyme in salty condition were identified and the corresponding gene was analyzed. Materials and methods: Strains from Urmia, Inche boron and Gomeshan Ponds were inoculated in the media containing pectin precursors. By analyzing the clear zones around the colonies based on I2/KI indicator, the positive strains were selected. Quantification of enzyme activity on all three types of pectinase was carried out by spectrophotometry. In order to molecularly screen the bacterium contained pectinase gene, the bacterium genome was amplified using appropriate primers. Results: Seventeen positive strains for pectinase (10 from Gomeshan Lake, 6 from Inche Boron Lake and 1 from where Urmia Lake) were identified among 130 studied samples. According to size of clear zone of enzyme activity in the qualitative test, activity level of all three pectinase enzymes in R2S25 strain of Inche Boron Lake was measured and the growth curve was obtained. Molecular study showed that all strains contain desired gene segment. Discussion and conclusion: Quantitative evaluation showed that production and activity of pectinase enzyme in R2S25 strain increased simultaneously with increasing the growth of selected strain in logarithmic phase. Molecular study also showed that the genuses of Martelella, Aeromocrobium, Planococcus, Marinobacter, Virgibacillus, Kocuria and Micrococcus contain the pectinase gene.http://bjm.ui.ac.ir/article_22300_0e7a5768e40e85fd9e36361895849738.pdfPectinase Enzymehalophilic bacteriaMolecular sieving |
| spellingShingle | Zohre Nasrollahzadeh Eniseh Saleh-Ghamari Mohammad Tahmaseb Mohammad Ali Amoozegar Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran Biological Journal of Microorganism Pectinase Enzyme halophilic bacteria Molecular sieving |
| title | Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran |
| title_full | Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran |
| title_fullStr | Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran |
| title_full_unstemmed | Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran |
| title_short | Bioassay and Molecular Screening of Pectinase Enzyme in halophilic bacteria from Salt Lake, Iran |
| title_sort | bioassay and molecular screening of pectinase enzyme in halophilic bacteria from salt lake iran |
| topic | Pectinase Enzyme halophilic bacteria Molecular sieving |
| url | http://bjm.ui.ac.ir/article_22300_0e7a5768e40e85fd9e36361895849738.pdf |
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