Topraktan fitaz enzimi üreten Bacillus sp. suşlarının taranması ve besinsel faktörlerin fitaz üretimi üzerine etkisi
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Date
2015-01-20
Authors
Baygın, Eren
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Publisher
Uludağ Üniversitesi
Abstract
Bu çalışmada Türkiye' nin 30 farklı ilinden temin edilen toprak örneklerinden 300 adet bakteri izole edilmiştir. Bakterilerin morfolojik ve fizyolojik özellikleri incelenmiş ve 236 bakteri Bacillus cinsi olarak tanımlanmıştır. Bütün suşların fitaz aktiviteleri fitaz tarama ortamı (PSM) kullanılarak tespit edilmiş ve açık zonların çapı mm olarak gösterilmiştir. Toplam 20 Bacillus sp. suşu ekstraselüler fitaz üreticisi olarak bulunmuştur. Bu suşlar arasında, en geniş zona (11 mm) sahip Bacillus sp. suşu seçilmiş ve bu suş Bacillus sp. EBD 9-1 olarak isimlendirilmiştir. Bu yeni izolat 6 farklı içerikli sıvı ortamda enzim üretim kapasitesi için test edilmiştir. Bu sıvı ortamlar arasında, en yüksek enzim aktivitesi (210 U/mL) TS ortamında tespit edilmiştir. Bacillus sp. EBD 9-1 suşunun 48. saate maksimum seviyede fitaz ürettiği bulunmuştur. Besinsel faktörler enzim üretimi üzerinde etkilidirler. Bu amaçla, enzim üretimi üzerine farklı karbon, nitrojen kaynakları ve metal iyonları gibi ana ortam maddelerinin etkileri araştırılmıştır. Kullanılan karbon kaynakları arasında, laktoz (408 U/mL) ve buğday kepeği (408 U/mL) enzim üretimi üzerinde çok yüksek bir potansiyel göstermiştir. En iyi organik azot kaynağı et özütü (365 U/mL) olarak bulunmuştur. İnorganik azot kaynakları organik kaynaklar kadar etkili değildiler. En iyi metal iyonu olarak CaCl2 (282 U/mL) bulunmuştur. Enzim üretimi, sırasıyla laktoz ve buğday kepeği, et özütü, CaCl2 varlığında % 94,% 74 ve% 34 oranında artmıştır. Fitaz üretimini artırmak amacıyla, yeni ortam en iyi karbon, nitrojen kaynakları ve metal iyonlarının birleştirilmesi ile elde edilmiştir. Bu ortamda, enzim verimi (311 U/mL), bazal ortamına kıyasla % 48 artırılmıştır. Bu yeni izole edilen suşun endüstriyel ölçekte fitaz üretimi, büyük bir potansiyele sahip olabilir.
In this study, 300 bacteria were isolated from soil samples that provided from 30 different cities of Turkey. Morphological and physiological features of bacteria were investigated, and 236 bacteria were determined as Bacillus genus. The phytase activities of all strains were assayed using phytase screening medium (PSM), and exhibited as diameter of clear zone in mm. Total 20 Bacillus sp. strains were found as extracellular phytase producer. The bacteria which has the most largest zone (11 mm) selected, and strain is named as Bacillus sp. EBD 9-1. This new isolate was tested for enzyme production capacity in the six different liquid media. Among of these liquid media, the highest enzymatic activity (210 U/mL) was determined in TS medium. It was found out that Bacillus sp. EBD 9-1 strain produced phytase on maximum level at 48th hour. The nutritional factors are affecting on the production of enzymes. For this purpose, the effects of major medium ingredients, such as different carbon, nitrogen sources and metal ions, on the production of the enzyme were investigated. Among the carbon sources used, lactose (408 U/mL) and wheat bran (408 U/mL) showed the highest potential on the enzyme production. The best organic nitrogen source was meat extract (365 U/mL). Inorganic nitrogen sources were not as effective as organic sources. CaCl2 (282 U/mL) was found as the best metal ions. The enzyme production was increased by 94% , 74% and 34% in the presence of lactose and wheat bran, meat extract CaCl2, respectively. In order to enhance the production of phytase, a new medium was obtained by combining the best carbon and nitrogen sources, and metal ions. In this medium, enzyme yield (311 U/mL) was enhanced 48% compared to basal medium. This newly isolated Bacillus sp. EBD 9-1 strain may be great potential for phytase production at industrial scale.
In this study, 300 bacteria were isolated from soil samples that provided from 30 different cities of Turkey. Morphological and physiological features of bacteria were investigated, and 236 bacteria were determined as Bacillus genus. The phytase activities of all strains were assayed using phytase screening medium (PSM), and exhibited as diameter of clear zone in mm. Total 20 Bacillus sp. strains were found as extracellular phytase producer. The bacteria which has the most largest zone (11 mm) selected, and strain is named as Bacillus sp. EBD 9-1. This new isolate was tested for enzyme production capacity in the six different liquid media. Among of these liquid media, the highest enzymatic activity (210 U/mL) was determined in TS medium. It was found out that Bacillus sp. EBD 9-1 strain produced phytase on maximum level at 48th hour. The nutritional factors are affecting on the production of enzymes. For this purpose, the effects of major medium ingredients, such as different carbon, nitrogen sources and metal ions, on the production of the enzyme were investigated. Among the carbon sources used, lactose (408 U/mL) and wheat bran (408 U/mL) showed the highest potential on the enzyme production. The best organic nitrogen source was meat extract (365 U/mL). Inorganic nitrogen sources were not as effective as organic sources. CaCl2 (282 U/mL) was found as the best metal ions. The enzyme production was increased by 94% , 74% and 34% in the presence of lactose and wheat bran, meat extract CaCl2, respectively. In order to enhance the production of phytase, a new medium was obtained by combining the best carbon and nitrogen sources, and metal ions. In this medium, enzyme yield (311 U/mL) was enhanced 48% compared to basal medium. This newly isolated Bacillus sp. EBD 9-1 strain may be great potential for phytase production at industrial scale.
Description
Keywords
Bacillus, Fitaz, Taranma, Besinsel faktörler, Phytase, Screening, Nutritional factors
Citation
Baygın, E. (2015). Topraktan fitaz enzimi üreten Bacillus sp. suşlarının taranması ve besinsel faktörlerin fitaz üretimi üzerine etkisi. Yayınlanmamış yüksek lisans tezi. Uludağ Üniversitesi Fen Bilimleri Enstitüsü.