Özet:
Bu çalısmada özellikle sarap ve sirkeden, ayrıca elma ve üzüm suyundan, selüloz
üreten bakteriler izole edilmistir. Bu izolatlardan selüloz üretim verimi en yüksek
olan iki izolat seçilmis ve bu izolatlar, klasik ve moleküler taksonomiye göre
Acetobacter pasteurianus HBB6 ve Acetobacter lovaniensis HBB5 olarak
tanılanmıstır. Bu strainlerin, karbon ve azot kaynakları, inkübasyon sıcaklıkları ve
pH’ ları degistirilerek selüloz üretim verimleri optimize edilmistir. Mikrobiyoloji
kültür koleksiyonu DSMZ (Almanya Mikroorganizmalar ve Hücre Kültürü
Koleksiyonu)’den alınan Gluconacetobacter xylinus DSM 46604,
Gluconacetobacter xylinus DSM 2004 ve Acetobacter aceti DSM 3508 strainleri
ile Acetobacter pasteurianus HBB6 ve Acetobacter lovaniensis HBB5
strainlerinin, optimum sartlardaki selüloz üretim verimleri melas, peynir altı suyu,
zeytin karasuyu ve corn steep liquor gibi ucuz atık maddeler kullanılarak
incelenmis ve sonuçlar karsılastırılmıstır. Selüloz üretiminde en iyi karbon
kaynagının glikoz, en iyi azot kaynagının ise yeast ekstrakt oldugu belirlenmistir.
Ayrıca sıcaklık ve pH denemeleri sonucunda; en iyi sıcaklık derecesinin 300C, en
iyi pH’ın da 6,5 oldugu saptanmıstır. Kullanılan atık maddeler içerisinde melas,
peynir altı suyu ve corn steep liquor içerikli besiortamlarında hücre gelisimi ile
birlikte selüloz üretimi gözlenmistir. Sadece zeytin karasuyunda hiçbir
mikroorganizma gelisimi olmamıs ve buna baglı olarak da selüloz üretimi
gerçeklesmemistir. Bu çalısmada, kullanılan tüm besiortamlarında, en yüksek
selüloz üretim veriminin, Gluconacetobacter xylinus DSM 46604 straininde, en
düsük selüloz üretim veriminin ise Acetobacter aceti DSM 3508 ve Acetobacter
lovaniensis HBB5 strainlerinde oldugu belirlenmistir. Acetobacter pasteurianus
HBB6 ve Acetobacter lovaniensis HBB5 strainlerinin morfolojik görüntüleri ile,
optimum sartlarda elde edilen selülozun agsı yapısı SEM’de görüntülenmistir.
Aynı zamanda, bakteriyal selülozun, enzimatik ve TFA ile asidik hidrolizinin
TLC analizi sonucunda, monosakkarit içeriginin glukoz oldugu belirlenmistir.
Ayrıca, bakteriyal selülozun NMR ve FT-IR spektrofotometre analizleri ile
kimyasal yapısı incelenmistir.
In the present study, cellulose producing bacteria were isolated mainly from wine
and vinegar as well as apple juice and grape juice. Two strains having highest
bacterial cellulose productivity were selected and these isolates were identified as
Acetobacter pasteurianus HBB6 and Acetobacter lovaniensis HBB5 using
classical and molecular taxonomy methods. In order to optimize the yield of
cellulose production of these strains, carbon and nitrogen sources, incubation
temperatures and pH conditions were adjusted. In addition, cellulose producing
output of Gluconacetobacter xylinus DSM 46604, Gluconacetobacter xylinus
DSM 2004 and Acetobacter aceti DSM 3508 strains, which were taken from
German Collection of Microorganisms and Cell Cultures, as well as Acetobacter
pasteurianus HBB6 and Acetobacter lovaniensis HBB5 were examined by using
some cheap waste substances like molasses, whey, olive oil black water and corn
steep liquor and the results were compared.
According to the results of this study, the best carbon and nitrogen sources are
glucose and yeast extract, respectively. On the other hand, 30 0C and pH 6.5 was
found optimal for the highest yield of cellulose production. Among the waste
substances examined, cellulose production along with growth of microorganism
was observed in the media containing molasses, whey and corn steep liquor. But,
no microorganism growth was detected only in use of black water of olive oil, so
cellulose production did not take place. In this work, the highest cellulose
producing output were found in Gluconacetobacter xylinus DSM 46604 strain
while Acetobacter aceti DSM 3508 and Acetobacter lovaniensis HBB5 were the
lowest cellulose producers in the all media examined.
Morphological view of Acetobacter pasteurianus HBB6 and Acetobacter
lovaniensis HBB5 strains and the bundle structure of cellulose which produced at
optimum conditions were monitored by scanning electron microscopy. At the
same time, as a result of enzymatic hydrolyse and acidic hydrolyse by TFA and
TLC analyze, glucose was found as the main content of bacterial cellulose
monosaccharide. Besides, the chemical structure of bacterial cellulose was
examined by NMR and FT-IR spectrophotometer.