Biochemical Studies Of Secondary Metabolite Of Penicillium Chrysogenum Grown On Selected Agro-Wastes

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Date
2012
Authors
ONYEGEME-OKERENTA, B.M.
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Journal ISSN
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Publisher
School of Postgraduate Studies University of Lagos
Abstract
Selected agro-wastes found in Lagos, Nigeria (cassava peels, corncob, sawdust, and sugarcane pulp) were compared with glucose and lactose as microbial substrates for cultivating P. chrysogenum (wild strain). This study was designed to give added value to agro waste as substrates to cultivate P. chrysogenum and subsequent production of secondary metabolite with antibiotic and anticoagulant properties. In the growth studies, corn cob and cassava peels significantly (p < 0.05) produced the highest amount of mycelia weight. Corn cob yielded a mycelia weight of 0.15 ± 0.02 and 0.92 ± 0.04mg/ml on the third and ninth day respectively while cassava peels yielded a mycelia weight of 0.13 ± 0.07 and 0.12 ± 0.02mg/ml on the third and twelfth day respectively. Mycelia weight of the organism, in media containing glucose, sugar cane and lactose was 0.12 ± 0.02, 0.068 ± 0.05 and 0.055 ± 0.03mg/ml respectively, was highest on the ninth day. Sawdust gave the least growth with a mycelia weight of 0.07 ± 0.01mg/ml on the third day. Cassava media has the highest carbohydrate content. Changes in extracellular protein secreted into the different media (every 3days for 21days) shows that culture media containing cassava peels gave the highest protein peak of 0.38 ± 0.08 mg/ml on the sixth day, while corncob gave an early peak of 0.30 ± 0.03mg/ml on the third day. Sawdust gave two protein peaks, 0.15 ± 0.03 on the third day and 0.25 ± 0.01mg/ml on the twelfth day. A total protein yield of 0.2 ± 0.05, 0.08 ± 0.02, 0.06 ± 0.02 mg/ml respectively was obtained with glucose, sugarcane pulp and lactose containing media on the third day. The results suggest that cassava peels, corncob and sugarcane pulp could serve as cheap fermentation substrates for the growth of the fungus. Optimum pH and temperature of growth and antibiotic production was 6.5 and 25OC respectively. elected agro-wastes found in Lagos, Nigeria (cassava peels, corncob, sawdust, and sugarcane pulp) were compared with glucose and lactose as microbial substrates for cultivating P. chrysogenum (wild strain). This study was designed to give added value to agro waste as substrates to cultivate P. chrysogenum and subsequent production of secondary metabolite with antibiotic and anticoagulant properties. In the growth studies, corn cob and cassava peels significantly (p < 0.05) produced the highest amount of mycelia weight. Corn cob yielded a mycelia weight of 0.15 ± 0.02 and 0.92 ± 0.04mg/ml on the third and ninth day respectively while cassava peels yielded a mycelia weight of 0.13 ± 0.07 and 0.12 ± 0.02mg/ml on the third and twelfth day respectively. Mycelia weight of the organism, in media containing glucose, sugar cane and lactose was 0.12 ± 0.02, 0.068 ± 0.05 and 0.055 ± 0.03mg/ml respectively, was highest on the ninth day. Sawdust gave the least growth with a mycelia weight of 0.07 ± 0.01mg/ml on the third day. Cassava media has the highest carbohydrate content. Changes in extracellular protein secreted into the different media (every 3days for 21days) shows that culture media containing cassava peels gave the highest protein peak of 0.38 ± 0.08 mg/ml on the sixth day, while corncob gave an early peak of 0.30 ± 0.03mg/ml on the third day. Sawdust gave two protein peaks, 0.15 ± 0.03 on the third day and 0.25 ± 0.01mg/ml on the twelfth day. A total protein yield of 0.2 ± 0.05, 0.08 ± 0.02, 0.06 ± 0.02 mg/ml respectively was obtained with glucose, sugarcane pulp and lactose containing media on the third day. The results suggest that cassava peels, corncob and sugarcane pulp could serve as cheap fermentation substrates for the growth of the fungus. Optimum pH and temperature of growth and antibiotic production was 6.5 and 25OC respectively. UV xxx modification of parent strain produced two mutant strains with 70% increase in penicillin production. In vitro antibacterial activity of the culture extracts was tested against some clinical bacterial isolates, namely, B. subtilis, E. coli, P. mirabilis and P. aeruginosa. Commercial Benzyl Penicillin was used as reference drug. The culture extracts and standard drug inhibited the growth of B. subtilis and E. coli. Zone of inhibition varied with the carbon source. Culture extracts and reference drug were not effective against the isolates of P. aeruginosa and P. mirabilis because they produce β-lactamase enzymes which hydrolyse the β-lactam present in the extract and reference drug. Antibacterial activity of extracts from cultures containing cassava peels and sugarcane pulp compared positively with that of the standard drug. The Minimum inhibitory concentration (MIC) of the reference drug against the susceptible organisms was 0.2 - 0.4mg/ml. For the culture extracts, the MIC ranged from 0.4 to 2.0mg/ml. It was 0.4 - 0.8mg/ml for cassava peels and sugarcane pulp, 0.6 - 0.8mg/ml for glucose and lactose, 0.8 - 1.0mg/ml for corncob and 1.0 - 2.0mg/ml for sawdust. Toxicity study showed that the extract is safe for use as there were no visible changes or recorded deaths 48 - 72hours after administration of the extracts. Haematological evaluation showed a significant decrease (p<0.05) in platelet count for both the extract and reference drug in the sub acute toxicity study as well as in infection and inflammatory conditions. The extract was shown to have a potent antithrombic and anticoagulant activities against thrombin and whole blood respectively. Higher concentrations of the extract and reference drug caused an increase in whole blood clotting time. At 6mg/ml of the extract and 5mg/ml of the reference drug, clot formation was not observed. There was complete inhibition of thrombin coagulation at concentrations above 10 and 20mg/ml of the reference drug and extract respectively
Description
Keywords
Agro-wastes , cassava peels , chrysogenum (wild strain) , inflammatory conditions , Haematological evaluation , Corn cob
Citation
Onyegeme-Okerenta, B.M (2012) Biochemical Studies Of Secondary Metabolite Of Penicillium Chrysogenum Grown On Selected Agro-Wastes. A Thesis Submitted to University of Calabar School of Postgraduate Studies Phd Thesis and Dissertation, 229pp.