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Register a new userProduction of B-galactosidase enzyme from some Microorganisms
إنتاج إنزيم B - غالاكتوزيداز B-galactosidase من بعض الأحياء الدقيقة
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Damascus University رسالة ماجستير
Publication date
2015
fields
Agricultural Engineering
and research's language is
العربية
Created by
Shamra Editor
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يعد إنزيم البتا - غالاكتوزيداز galactosidases (B-Ec 3.2.1.23) من أهم الإنزيما المستخدمة في الصناعات الغذائية وله العديد من التطبيقات الغذائية والتكنولوجية والبيئية الهامة في مجلات الحياة المختلفة . ويعتبر مثالاً هاماً لإنزيمات الحلمهة حيث يقوم البتا- غالاكتوزيداز بحلمهة اللاكتوز غالاكتوزيداز إلى ىسكريات أحادية وهي الغلوكوز والغلاكتوز .
B-D-Galactosidase (B-D- galactoside galactohydrolase , E. C. 3.2.1.23) , is one of the most important enzymes used in food processing . It a specific example of a glycosidase enzyme. Many organisms naturally synthesize B- galactosidase , including microorganisms , plants and animal cells . Among these possibilities , microbial sources are the best.
References used
ِAdam, A.c. Ruhio- Texeira , M. Polaina . J (2004) Lactose : the milk sugar from a biotechnological perspective
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40 samples were collected from milk from Homs to isolate the
bacteria and study their ability to produce the enzyme β-
galactosidase and diagnose isolates the most efficient in the
production of the enzyme, the results showed the presence of 25
sporeforming isolates producing the enzyme β-galactosidase , the
effectiveness of β-galactosidase was estimated using
spectrophotometer at a wavelength of 420 nm.
β-galactosidase enzyme was isolated from the new born goat brain by nine
methods, It was found that the sodium acetate 0.2 Mole/Liter +0.2Mole/Liter
NaCl PH5 method have given the highest specific activity of crude enzyme in
comparison with the ot
her methods. Also, this enzyme was purified by using
four methods, the second one (cold acetone) was the butter. As a result the
purification fold was about 135.46 times and the yield about 77.14% by using
Sephacryl S200 (second step). This enzyme is 187.437 KDa as a molecular
weight.
Some characteristics of β-galactosidase enzyme that was isolated from a new
born goat brain were studied. This study concluded that the enzyme is glucoenzyme
in which the carbohydrate part constitutes 22.1% in accordance with
phenol –sulfate acid
method.
The optimum pH for the enzyme activity is 5.5. The enzyme lost its activity
completely at pH8.5, and showed great stability at the range of pH 4-6.
The results indicated that the optimum temperature for the enzyme activity
is 55Co at the optimum pH. The stability temperature for the enzyme is
35-60Co.
The analytical results of 5%lactose solution hydrolyzed by the enzyme have
indicated that the hydrolysis rate is between 40% after 60 minutes, to 95%
after 270 minutes.
Forty Isolates of Streptomyces bacteria were obtained from soil samples
from different regions of Syria (Damascus, Sweida, Daraa, Homs, Latakia,
Damascus, Hasakeh, Deir-ezzor) during 2012. The isolates were identified using
ISP (International Stre
ptomyces Project) methodology based on morphological
and physiological criteria. Isolates were grown on selective solid media
depending on growth density as an indicator of the consumption of xylose as an
unique carbon source then on liquid media to determine their ability to
produce the glucose isomerase enzyme. The crude enzyme was extracted using
CTAB 0.1% solution, the enzyme activity was estimated using glucose as a
substrate and the end product was measured colorimetrically at 560 nm.
Results showed that the 40 isolates belong to 29 different species of
Streptomyces. All isolates were able to produce glucose isomerase but they
varied in their efficiency. Isolate SH10 (S. roseiscleroticus) was distinguished by
its high level of enzyme production reaching 4.9 units /ml.
Bioeffect of some Syrian Myrtus communis L. leaves extracts in growth of some pathogenic microorganisms, which were isolated from Al-Assad hospital laboratory in Lattakia, was tested by disc diffusion method. Results showed that cold and hot water ex
tracts have antibacterial activity against all Gram positive and negative bacteria, and pathogenic Candida albicans except Klebsiella pneumoniae. All organic extracts have antibacterial activity against Proteus vulgaris, Entrococcus faecium, Staphylococcus aureus and Staphylococcus albus.
Pseudomonas aeruginosa and Pseudomonas fluorescens were affected by most of the organic extracts, and the most bioactivity was by methanol extract, and all inhibition zones of extracts were bigger than inhibition zones by control
antibiotics. Serratia marcescens and Acinetobacter calcoaceticus were affected by methanol (with highest inhibition zone 37.33 and 27.33 mm respectively), ethanol, acetone and ethyl acetate extracts, but resistant to the others. Klebsiella pneumoniae affected by ethanol extract only with inhibition zone 20.33 mm.
Results of bioactivity according to a various solvents used in this study demonstrate that Syrian Myrtus communis L. leaves extracts have antimicrobial activity, so expected to be potential sources of natural antibacterial and antifungal products against some pathogenic bacteria and fungi in the future.
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