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Paper Information

Journal:   BIOLOGICAL JOURNAL OF MICROORGANISM   SUMMER 2018 , Volume 7 , Number 26 ; Page(s) 115 To 122.
 
Paper: 

BIOASSAY AND MOLECULAR SCREENING OF PECTINASE ENZYME IN HALOPHILIC BACTERIA FROM SALT LAKE, IRAN

 
 
Author(s):  NASROLLAHZADEH ZOHRE, SALEHGHAMARI ENSIEH, TAHMASEB MOHAMMAD*, AMOOZEGAR MOHAMMAD ALI
 
* DEPARTMENT OF CELL AND MOLECULAR SCIENCES, FACULTY OF BIOLOGICAL SCIENCES, KHARAZMI UNIVERSITY, TEHRAN, IRAN
 
Abstract: 

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.

 
Keyword(s): PECTINASE ENZYME, HALOPHILIC BACTERIA, MOLECULAR SIEVING
 
References: 
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