Abstract
Seventeen local isolates of Bacillus were isolated from soil to produce extracellular xylanase under submerged fermentation process by using xylan as carbon sole source. All isolates were subjected to quantitative scanning to select the most efficient one. The highest activity of xylanase (2680u/ml) was obtained from isolate Bacillus sp RS1. The isolate identified by 16SrRNA gene sequence of Bacillus subtilis ( accuracy of 99%)which was matched with sequence of Bacillus subtilis VBN25 that recorded in Genebank under the Accession Number of MG027675.1.Extracted xylan from agricultural waste by acidic method(papyrus, sun flower stalks, Ibaa Wheat type, Furat wheat type and Abo Ghraib wheat type)were used as the substrate for xylanase production from Bacillus. The results showed that the papyrus gave the highest amount of xylan (187.6 µg/ml) as compared with that of the sun flower stalks, Ibaa Wheat type, Furat wheat type and Abo Ghraib wheat type(161.3, 161.6, 157.6, 157.2) µ g/ml respectively. The results indicated that the highest xylanase activity was 2800 u/ml produced by Bacillus subtilus when Papyrus xylan was used.
Highlights
Many bacterial genera including Bacillus, Cellulomonas, Clostridium, Rumminococcus, Acetivibrio, Bacteriodes, and Alteromonas can produce xylanas (17).Bacillus subtilis is nonpathogenic and nontoxigenic bacterium that has been used a source of enzymes (19)
Identification of Bacillus isolates Study of cultural and morphological characteristics of Bacterial isolates The cultural characteristics of bacterial isolates on NA media and morphological characteristics, was carried out by using Gram and malachite stains according to method of (7)
After the fermentation process was finished, the supernatant was separated by centerfuge at aspeed 12000 rpm for 10 min at 4co (13).The supernatant was used as the source of crude enzyme for xylanase assay
Summary
Many bacterial genera including Bacillus, Cellulomonas, Clostridium, Rumminococcus, Acetivibrio, Bacteriodes, and Alteromonas can produce xylanas (17).Bacillus subtilis is nonpathogenic and nontoxigenic bacterium that has been used a source of enzymes (19). The aim of this study is to get Bacillus isolate from local sources which produce a large amounts of xylanase enzyme by using agricultural waste. Screening the isolates for xylanase production Production media was placed in to 250 ml Erlenmeyer flask, autoclaved at 121co for 15 min, Flasks were inoculated with 1ml cell suspension at adensity of 107 cell /ml media.The fermentation was carried out in ashaking incubater at aspeed of 150 rpm at 37 co for 48 hr (5). After the fermentation process was finished, the supernatant was separated by centerfuge at aspeed 12000 rpm for 10 min at 4co (13).The supernatant was used as the source of crude enzyme for xylanase assay. The device was programmed according to table (3) and the amplification was taken place to amplify the extracted DNA.
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