Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust

xvii,82

-д хадгалсан:
Номзүйн дэлгэрэнгүй
Үндсэн зохиолч: Agbolade, Abisola Ebunoluwa
Формат: Дипломын ажил
Хэл сонгох:англи
Хэвлэсэн: Microbiology, Obafemi Awolowo University 2020
Нөхцлүүд:
Онлайн хандалт:https://ir.oauife.edu.ng/handle/123456789/5151
Шошгууд: Шошго нэмэх
Шошго байхгүй, Энэхүү баримтыг шошголох эхний хүн болох!
_version_ 1810764570334068736
author Agbolade, Abisola Ebunoluwa
author_facet Agbolade, Abisola Ebunoluwa
author_sort Agbolade, Abisola Ebunoluwa
collection DSpace
description xvii,82
format Thesis
id oai:ir.oauife.edu.ng:123456789-5151
institution My University
language English
publishDate 2020
publisher Microbiology, Obafemi Awolowo University
record_format dspace
spelling oai:ir.oauife.edu.ng:123456789-51512023-05-13T11:29:12Z Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust Agbolade, Abisola Ebunoluwa Bacterium Isolated Decaying Sawdust Alcohol Production Fermentation Sawdust Cellulase Cellulase enzyme xvii,82 This study investigated fermentation of off-cut waste paper hydrolysed with cellulase enzyme produced by a bacterium isolated from decaying sawdust; isolated a cellulolytic bacterium; identified and optimized conditions for cellulase production by the isolate. This was with a view to hydrolysing off-cut waste paper with the cellulase enzyme obtained for bioethanol production. Samples of decaying sawdust were collected from a saw mill and were serially diluted. Aliquot of each dilution was appropriately plated and incubated. The bacterial isolates obtained from the plates were screened for cellulolytic ability on carboxymethylcellulose agar plates. The bacterial isolate with the highest cellulolytic activity was selected and identified by 16S rRNA gene sequencing. Conditions for cellulase production were optimized and cellulase was produced by submerged fermentation under the optimum conditions recorded. Partial purification of cellulase was achieved by ion exchange and gel filtration chromatography. The kinetics and other physicochemical parameters of cellulase were determined. Off-cut waste paper was soaked in distilled water to form a mash which was pretreated with alkaline, alkaline-peroxide and dilute acid pretreatment methods. The pretreated mashes were hydrolysed with both crude and purified cellulase, alongside untreated mash for 48 h and the reducing sugars released were monitored. The pH of the enzyme-hydrolysed mashes was adjusted to 5.5, sterilized, inoculated with standardized broth culture of 24 h old Saccharomyces cerevisiae and incubated at 30oC for 48 h. The mixture was analysed for alcohol production before and after distillation using acidified-dichromate method. Twenty-six (26) bacterial isolates were obtained. Seven (7) of the isolates (26.92%) tested positive for cellulose hydrolysis with appreciable halozones. The best bacterial isolate with a halozone of 58 mm and an initial cellulase activity of 29.36 Units/ml was identified as Bacillus subtilis A8. Optimum conditions for cellulase production by Bacillus subtilis A8 were determined to be a temperature of 35 ± 2oC, pH 7, 0.8% substrate concentration, 4% inoculum size, carboxymethylcellulose and peptone as the more preferred carbon and nitrogen source respectively. Enzyme yields of 18.5 and 13.5% were obtained from purification by ion exchange and gel filtration chromatography respectively. The Michealis-Menten (km) was found to be 0.0108 ± 0.0032 mg/ml with a maximum velocity (Vmax) of 119.3 ± 7.4 µmol/min. Maximum activity for partially purified cellulase was recorded at pH 9.5 and 55oC with stability at 50oC; and pH 9, 35oC with stability at 45oC for crude cellulase. Hydrolysis of dilute acid pretreated waste paper with crude cellulase gave the highest quantity of reducing sugars at the 48th hour, and also gave the highest ethanol yield upon fermentation and distillation. The study concluded that cellulase from Bacillus subtilis A8 could be utilized in the hydrolysis of waste paper to generate reducing sugars which can be fermented to bioethanol. Also, pretreatment of off-cut waste paper produced more ethanol. 2020-02-06T11:13:37Z 2020-02-06T11:13:37Z 2015 Thesis Agbolade, A.E. (2015). Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust. Obafemi Awolowo University https://ir.oauife.edu.ng/handle/123456789/5151 en application/pdf Microbiology, Obafemi Awolowo University
spellingShingle Bacterium Isolated
Decaying Sawdust
Alcohol Production
Fermentation
Sawdust
Cellulase
Cellulase enzyme
Agbolade, Abisola Ebunoluwa
Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
title Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
title_full Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
title_fullStr Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
title_full_unstemmed Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
title_short Fermentation of off-cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
title_sort fermentation of off cut waste paper hydrolysed with cellulase produced by a bacterium isolated from decaying sawdust
topic Bacterium Isolated
Decaying Sawdust
Alcohol Production
Fermentation
Sawdust
Cellulase
Cellulase enzyme
url https://ir.oauife.edu.ng/handle/123456789/5151
work_keys_str_mv AT agboladeabisolaebunoluwa fermentationofoffcutwastepaperhydrolysedwithcellulaseproducedbyabacteriumisolatedfromdecayingsawdust