Bioethanol Production with Cellulase Enzyme from Bacillus cereus Isolated from Sesame Seed Residue from the Jazan Region

Abada, Emad; Masrahi, Yahya S.; Alabboud, Mohamed A.; Alnashiri, Hassien M.; El-Gayar, Khaled E.

doi:10.15376/biores.13.2.3832-3845

Cited by 13 publications

(5 citation statements)

References 26 publications

(30 reference statements)

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“…Salah satu enzim yang memiliki peranan penting dalam pembuatan bioetanol adalah selulase (Abada et al, 2018). Harga selulase murni di pasaran cenderung sangat mahal, sehingga salah satu langkah alternatif untuk mendapatkan sumber selulase yang murah yaitu dengan memanfaatkan limbah rumen kambing yang diperoleh dari Rumah Potong Hewan (RPH).…”

Section: Pendahuluanunclassified

Enzimatic Kinetics of Cellulose Hydrolysis using Cellulase from Goat Rumen Fluids

Wijonarko

Purbowati²,

Maksum³

2022

ijft

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In line with the depletion of petroleum reserves from fossils, humans have forced people to look for alternative renewable energy sources. Nipah plant stems are organic material that contains a lot of lignocellulose. Cellulose in lignocellulose can be hydrolyzed to glucose to produce bioethanol. Problems arise because hydrolysis using pure cellulase requires relatively high costs, so it is necessary to find an alternative source of cheap cellulase. One of the cheap sources of cellulase is goat rumen fluid. Until now, there is not much information regarding the reaction kinetics and cellulase characteristics of goat rumen fluid. The purpose of this study was to isolate crude cellulase extract, determine the KM and Vmax values and determine the optimum temperature and pH. The research was carried out at the Unsoed Agricultural Technology Laboratory. The samples used were seven rumen fluids. The study began with isolation followed by enzymatic kinetics studies. Fractionation was carried out by adding ammonium sulfate salt (50, 60,70, and 80 %), and centrifugation at 7,000 rpm at 4 oC for 15 minutes. Crude cellulase extract with the highest enzyme activity value was used for the study of enzymatic reaction kinetics and characterization. The crude cellulase resistance test to temperature was carried out at 45, 55 and 65 oC and the pH was carried out at pH 4, 5, 6, 7 and 8. The KM and Vmax values were determined by measuring the activity of crude cellulase extract at various concentrations of CMC (1, 1 ,5, 2, 2.5, and 3 %). The results showed that crude cellulase extract had an average specific activity of 1.7356 IU/mg. The highest enzyme activity was 0.9854 IU/ml. The optimum crude cellulase extract was at pH 6 with an activity of 1.1015 IU/ml and a temperature of 60 oC with an activity of 0.7829 IU/ml. At a CMC concentration of 2.5% crude cellulase extract had an activity of 0.3179 IU/ml with a Vmax value of 0.0045 IU/ml and a KM of 0.0252 %.

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Section: Pendahuluanunclassified

Enzimatic Kinetics of Cellulose Hydrolysis using Cellulase from Goat Rumen Fluids

Wijonarko

Purbowati²,

Maksum³

2022

ijft

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“…The observation in this study was in consonance with the previous report of Hashem et al [44] that mentioned a species of Kluyveromyces was used as the fermenting yeast in the production of ethanol. Other authors such as Saliu and Sani [33] and Abada et al [45] employed Saccharomyces cerevisiae to carry out the fermentation of hydrolyzed cellulolytic biomass to ethanol.…”

Section: Pre-treatment and Saccharification Of Plant Wastes To Hydrolysable Sugarsmentioning

confidence: 99%

Saccharification and Fermentation of Cellulolytic Agricultural Biomass to Bioethanol using Locally Isolated Aspergillus niger S48 and Kluyveromyces sp. Y2, respectively

Adeyemo

Ja’afaru

Abdulkadir

et al. 2021

Period. Polytech. Chem. Eng.

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Due to increase in demand for energy as a result of human population explosion, industrialization and environmental hazards posed by fossil fuels, there is a need to source for alternative energy sources that are cheaper and environmental friendly. Three different lignocellulosic biomasses were studied for their suitability for bioethanol production. Fungi and yeasts were isolated using serial dilution and spread plate methods. Identification of both fungi and yeasts was done using their cultural and microscopy characteristics. Saccharification of the pre-treated biomass was done with both crude cellulase and mycelia inoculant. Bioethanol was produced using batch culture fermentation. Ethanol produced was detected using spectrometric method and quantified using High Performance Liquid Chromatography (HPLC). The effects of substrate concentration, pH and temperature on ethanol yield were optimized. Fifty fungal isolates were obtained from soil collected. Six yeasts, all Kluyveromyces species fermented three sugars to ethanol with isolate Kluyveromyces sp.Y2 having the shortest time. It was selected for fermentation. Aspergillus niger S48 had highest cellulase activity measured in a zone of hydrolysis of 26.0 mm. It had the highest glucanase activity, endoglucanase (0.462 U/mL) and exoglucanase (0.431 U/mL). The outcome of this study indicated that crude cellulase produced by Aspergillus niger S48 hydrolyzed the pre-treated rice chaff with 1.07 mg/mL of fermentable sugars higher than 0.87 mg/mL when the mycelia of the fungus was inoculated to pretreated rice chaff for hydrolysis. Ethanol was optimally produced at 12 % substrate concentration using rice chaff, at a temperature of 35 °C and pH 5.0.

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“…The highest amylase producing halophilic bacteria were identified by using 16S rRNA method according to Abada et al, 2018. [12] Total genomic DNA was isolated from the bacterial cells using phenol-chloroform method as described by Ruzzante et al, 1996. [13] Isolated DNA was checked for its quality and concentration by agarose gel electrophoresis using UV transilluminator.…”

Section: S Rrna Identification Of Halophilic Bacterial Isolatesmentioning

confidence: 99%

Molecular characterization of gene encoding halo tolerant amylase of Bacillus paralicheniformis isolated from Jazan region

Ghazouani

El-Gayar

Abada

2020

MJMB

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Objectives: This study aims to characterize the gene encoding halo tolerant amylase of bacteria isolated from Jazan region. Materials and Methods: Soil samples were collected from several area of Jazan region, KSA. The samples were serially diluted and plateted on starch agar plates. The amylase producing bacteria were detected by iodine test. To determine the halophilic amylase producing bacteria, several colonies were tested for their ability to grow at higher concentrations of NaCl ranging from 7 to 16%. The bacteria was identified by 16S rRNA and the full length amylase gene was fully identified by sequencing using specific primers. Results: One bacterial halophilic isolate was able to grow on starch agar medium up to 14% NaCl. The Gram stain of the isolate indicated that it is Gram-positive, bacilli. The 16S rRNA gene homology study showed that the bacterial isolate was identified as Bacillus paralicheniformis. Two specific primers were designed named S1F, S1R, to amplify the amylase gene (AMY) region using PCR and the PCR product was sequenced. The sequencing results showed that the full-length amy gene of B. paralicheniformis was of 1452 encoding 483 amino acids. The expected M.Wt. of the protein expressed is of 55 KDa. Conclusion: We report the isolation, identification, and characterization of an isolate of halophilic bacterium isolated from Jazan region. Based on molecular identification, this isolate was identified as Bacillus paralicheniformis. This bacterial strain has an α-amylase gene in its genome and is able to produce extracellular α-amylase. Based on the findings of this work we propose that Bacillus paralicheniformis amy gene could be cloned into expression vector for large scale production.

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Bioethanol Production with Cellulase Enzyme from Bacillus cereus Isolated from Sesame Seed Residue from the Jazan Region

Cited by 13 publications

References 26 publications

Enzimatic Kinetics of Cellulose Hydrolysis using Cellulase from Goat Rumen Fluids

Enzimatic Kinetics of Cellulose Hydrolysis using Cellulase from Goat Rumen Fluids

Saccharification and Fermentation of Cellulolytic Agricultural Biomass to Bioethanol using Locally Isolated Aspergillus niger S48 and Kluyveromyces sp. Y2, respectively

Molecular characterization of gene encoding halo tolerant amylase of Bacillus paralicheniformis isolated from Jazan region

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