Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province
The study included isolating and diagnosing of Bacillus subtilis different soils at Thi Qar province. The ability of bacterial isolates to produce Endo-β-1,4-glucanase was determined after growing on carboxymethyl cellulose CMC medium, Using Congo red and NaCl. Enzyme Endo-β-1,4-glucanase is...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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University of Thi-Qar
2019-04-01
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| Series: | مجلة علوم ذي قار |
| Subjects: | |
| Online Access: | http://jsci.utq.edu.iq/index.php/main/article/view/71 |
| _version_ | 1827595308027084800 |
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| author | Mohammed Ali Kassab Rahman Laibi Chelab |
| author_facet | Mohammed Ali Kassab Rahman Laibi Chelab |
| author_sort | Mohammed Ali Kassab |
| collection | DOAJ |
| description | The study included isolating and diagnosing of Bacillus subtilis different soils at Thi Qar province. The ability of bacterial isolates to produce Endo-β-1,4-glucanase was determined after growing on carboxymethyl cellulose CMC medium, Using Congo red and NaCl. Enzyme Endo-β-1,4-glucanase isolates were identified using biochemical tests and VITEK2 as B. subtilis. Isolates were identified using 16S rRNA tests after extraction of DNA from isolates and amplified by PCR using 27F primers (Forward) and 1492R (Reverse) . All isolates in the current study were positive for this gene and the size of the gene for all isolates was at 1500 pb. After identifying the gene sequences and comparing them with the data available in the Gen Bank, NCBI data showed that they were new strains of B. subtilis strain m1 (MF449304) and B. subtilis strain M2 (MF449461) bacteria. Isolates were recorded in NCBI GenBank and were design for each evolutionary tree isolation by Software MEGA6. After the diagnosis of bacterial isolates, the ideal conditions for the production of Endo-β-1,4-glucanase were changed for incubation period, temperature, pH , Incubator Shake, carbon and nitrogen sources. Enzymatic efficacy was determined using of dinitro salicylic acid DNS detector to detect the glucose releasing glucose molecules Endo-β-1,4-glucanase. The enzyme was produced after growing of bacterial isolates of plant culture containing plant and cardboard residues as natural sources of carbon and alternative to costly industrial sources as a source of carbon at a concentration of 1% at 45 ° C for 48 hours and pH (6 - 5) The highest wheat bran and millet bran has superior efficacy over other natural and industrial sources were used as an alternative to the industrial nitrogen source of peptone with a concentration of 1%. The malt extract as the best nitrogen source for all isolates in the present study was superior to all other nitrogen sources. Other sources came from either banana Peel that showed no growth for all isolates.
The study aimed to isolate and diagnose B. subtilis producing an Endoglucanases using cheap natural sources as alternatives source to reduce to costly carbon cost of and nitrogen sources and benefiting from biological treatment of agricultural and industrial waste. |
| first_indexed | 2024-03-09T02:51:45Z |
| format | Article |
| id | doaj.art-f961b05f95c84eaca636ce08c07b9c46 |
| institution | Directory Open Access Journal |
| issn | 1991-8690 2709-0256 |
| language | English |
| last_indexed | 2024-03-09T02:51:45Z |
| publishDate | 2019-04-01 |
| publisher | University of Thi-Qar |
| record_format | Article |
| series | مجلة علوم ذي قار |
| spelling | doaj.art-f961b05f95c84eaca636ce08c07b9c462023-12-05T09:58:18ZengUniversity of Thi-Qarمجلة علوم ذي قار1991-86902709-02562019-04-0164Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar provinceMohammed Ali Kassab0Rahman Laibi Chelab1Biology Department/ College of Education for pure science/Thi-Qar UniversityBiology Department/ College of Education for pure science/ Thi-Qar UniversityThe study included isolating and diagnosing of Bacillus subtilis different soils at Thi Qar province. The ability of bacterial isolates to produce Endo-β-1,4-glucanase was determined after growing on carboxymethyl cellulose CMC medium, Using Congo red and NaCl. Enzyme Endo-β-1,4-glucanase isolates were identified using biochemical tests and VITEK2 as B. subtilis. Isolates were identified using 16S rRNA tests after extraction of DNA from isolates and amplified by PCR using 27F primers (Forward) and 1492R (Reverse) . All isolates in the current study were positive for this gene and the size of the gene for all isolates was at 1500 pb. After identifying the gene sequences and comparing them with the data available in the Gen Bank, NCBI data showed that they were new strains of B. subtilis strain m1 (MF449304) and B. subtilis strain M2 (MF449461) bacteria. Isolates were recorded in NCBI GenBank and were design for each evolutionary tree isolation by Software MEGA6. After the diagnosis of bacterial isolates, the ideal conditions for the production of Endo-β-1,4-glucanase were changed for incubation period, temperature, pH , Incubator Shake, carbon and nitrogen sources. Enzymatic efficacy was determined using of dinitro salicylic acid DNS detector to detect the glucose releasing glucose molecules Endo-β-1,4-glucanase. The enzyme was produced after growing of bacterial isolates of plant culture containing plant and cardboard residues as natural sources of carbon and alternative to costly industrial sources as a source of carbon at a concentration of 1% at 45 ° C for 48 hours and pH (6 - 5) The highest wheat bran and millet bran has superior efficacy over other natural and industrial sources were used as an alternative to the industrial nitrogen source of peptone with a concentration of 1%. The malt extract as the best nitrogen source for all isolates in the present study was superior to all other nitrogen sources. Other sources came from either banana Peel that showed no growth for all isolates. The study aimed to isolate and diagnose B. subtilis producing an Endoglucanases using cheap natural sources as alternatives source to reduce to costly carbon cost of and nitrogen sources and benefiting from biological treatment of agricultural and industrial waste.http://jsci.utq.edu.iq/index.php/main/article/view/71: B. subtilis strain m1M2Produce Endo-β-14-glucanasecarbon sourcenitrogen source and DNS Reagent |
| spellingShingle | Mohammed Ali Kassab Rahman Laibi Chelab Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province مجلة علوم ذي قار : B. subtilis strain m1 M2 Produce Endo-β-1 4-glucanase carbon source nitrogen source and DNS Reagent |
| title | Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province |
| title_full | Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province |
| title_fullStr | Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province |
| title_full_unstemmed | Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province |
| title_short | Detection of local Bacillus subtilis producing Endo-β-1,4-glucanase Of Thi Qar province |
| title_sort | detection of local bacillus subtilis producing endo β 1 4 glucanase of thi qar province |
| topic | : B. subtilis strain m1 M2 Produce Endo-β-1 4-glucanase carbon source nitrogen source and DNS Reagent |
| url | http://jsci.utq.edu.iq/index.php/main/article/view/71 |
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