استخدام عوامل استحثاث لانتاج مؤيضات ثانوية من بعض عزلات انواع الفطر واختبار فعاليتها الضد بكتيرية والخلايا السرطانية Penicillium والكولسترول == Use of inducing agents in secondary metabolites production from some Penicillium isolates and assay their activity against bacteria , cancer cells and cholesterol
Author name:
امل صالح عبد الرضا
Supervisor name:
توفيق محمد محسن
General topic:
Biology
Specific topic:
Microbiology - Fungi
Degree:
Doctorate
University:
University Of Basrah - College Of Education For Pure Sciences
Language:
Arabic
University location:
Basrah
First pages:
24T3528 - p.pdf
Abstract:
The current study was designed to investigate active compounds from different Penicillium isolates ,that isolated from different sources , using different media . Eleven isolates related to five species : P. chrysogenum, P.paxilli , P. citrinum, P. cosmopolitanum, P. digitatum were isolated. After primary screening was performed , the best isolates that have high antibacterial activity against gram positive and gram negative bacterial was selected. The selected isolates were cultured on fermentation media according to the suitable media for each isolate, and the active compounds were extracted using ethyl acetate as a solvent. P. cosmopolitanum and P. chrysogenum were cultured on penicillin production medium (PPM) , P. paxilli was cultured on Aspergillus complete medium (ACM) and P. digitatum and P. citrinum were cultured on solid state fermentation media consist of wheat bran . The efficiency of fungal crude extracts from each isolate against the reference strains S. aureus NCTC 6571 and E. coli ATCC 25922 and against Candida albicans (pathogenic and reference strain)was examined, and the Minimum inhibitory concentration (MIC) was determinated for each extracts.The inhibition zone of fungal extracts against E. coli and S. aureus ranged between (20 - 50) mm , and they had high activity against C. albicans ( reference and pathogenic isolate) ranged between 25 - 45 mm.Induction of secondary metabolite production by the selected fungi was carried out using biological factor (Streptomyces sp.) , chemical agent (colchicine) and physical agent by UV radiation. Apparently that the biological induction by Streptomyces isolate was the best compared with the other treatments.The results revealed that the two fungal extracts ALO1 and Apax exhibit a high antioxidant activity at low concentrations and volumes of the extract compared to ascorbic acid as control. ALO1 had radical scavenging activity (RSA%) reach 99.9% at volume 3μl , and Apax had RSA 91.6% at concentration 500μg/ml. Also, the fungal extract ,ALO1, showed a significant activity as anticancer agent against RD - cell line. The IC50 of ALO1 was 8.5μl after 24h and 23.76μl after 72h. The hypercholesterolemia ability of ALO1 in vivo and in vitro was assayed . In vivo experiment designed using female albino mice and divided into three groups each one contain 8 mice : group 1 treated with 10μl , group 2 treated with 20μl of ALO1 , and group 3 treated with normal saline as control group. The results showed that total cholesterol , triglycerides, LDL and VLDL were significantly reduced when compared with control group while the HDL level was increased in comparing with control group. The chemical analysis of the crude extracts showed that Apen2 contained 259 μg/g of penicillin G when analyzed with HPLC apparatus . Also the GC mass analysis of ALO1 showed that Docasadionic acid initialize area76.18% and Palmitic acid initialize area 15.09% from the total identified compounds.Whereas the GC mass analysis of Apax showed that Hexandioic acid initialize area40% and Stearolic acid initialize area 35.57% from the total identified compounds.In addition , it was detected the presence of penicillin biosynthetic gene cluster : pcbAB, pcbC and penDE in P. cosmopolitanum and P. chrysogenum.The result showed that gene pcbC and penDE were found in the genomic DNA but the gene pcbAB was not amplified by PCR.
