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Effect of some environmental parameters on biobutanol production by Clostridium acetobutylicum NCIMB 13357 in date fruit medium

Khamaiseh EI, Hamid AA, Yusoff WM, Kalil MS

Pak J Biol Sci, 2013 Oct 15;16(20):1145-51.
PMID: 24506014

Date fruit juice contains high concentration of simple sugars ranging from 65 to 75% (w/w) in dry form. In this study, the potential of date fruit juice as biobutanol fermentation medium by C. acetobutylicum was investigated. The fermentation process was carried out at initial pH of 5, 6 and 7, incubation temperature of 30, 35 and 40 degrees C for 72 hours. The date fruit concentrations tested were 10, 20, 30 and 40 g L(-1). Medium containing 30 g L(-1) of date fruit at 35 degrees C incubation temperature with initial medium pH 7.0 gave the highest concentration of solvents of 3.1, 0.1 and 1.1 g L(-1) butanol, ethanol and acetone respectively. The yield and productivity of biobutanol were 0.32 g g(-1) and 0.044 g L(-1)/h respectively, while for total ABE were 0.45 g g(-1) and 0.06 g L(-1) h, respectively.

Matched MeSH terms: Clostridium acetobutylicum/growth & development*; Clostridium acetobutylicum/metabolism*
Fulltext Enhanced butanol production by Clostridium acetobutylicum NCIMB 13357 grown on date fruit as carbon source in P2 medium

Khamaiseh EI, Abdul Hamid A, Abdeshahian P, Wan Yusoff WM, Kalil MS

ScientificWorldJournal, 2014;2014:395754.
PMID: 24672315 DOI: 10.1155/2014/395754

The production of biobutanol was studied by the cultivation of Clostridium acetobutylicum NCIMB 13557 in P2 medium including date fruit as the sole substrate. The effect of P2 medium and the effect of different concentrations of date fruit ranging from 10 to 100 g/L on biobutanol production were investigated. Anaerobic batch culture was carried out at 35 °C incubation temperature and pH 7.0 ± 0.2 for 72 h. Experimental results showed that the lowest yield of biobutanol and acetone-butanol-ethanol (ABE) was 0.32 and 0.35 gram per gram of carbohydrate consumed (g/g), respectively, when an initial date fruit concentration of 10 g/L was utilized. At this fruit date concentration a biobutanol production value of 1.56 g/L was obtained. On the other hand, the maximum yield of biobutanol (0.48 g/g) and ABE (0.63 g/g) was produced at 50 g/L date fruit concentration with a biobutanol production value as high as 11 g/L. However, when a higher initial date fruit concentration was used, biobutanol and ABE production decreased to reach the yield of 0.22 g/g and 0.35 g/g, respectively, where 100 g/L date fruit was used. Similar results also revealed that 10.03 g/L biobutanol was produced using 100 g/L date fruit.

Matched MeSH terms: Clostridium acetobutylicum/metabolism*
Effect of some environmental parameters on hydrogen production using C. acetobutylicum

Alshiyab H, Kalil MS, Hamid AA, Yusoff WM

Pak J Biol Sci, 2008 Sep 01;11(17):2073-82.
PMID: 19266920

The aim of this study was to investigate the influence of some environmental factors on bacterial metabolism. Fermentative hydrogen production by C. acetobutylicum, using glucose as the substrate. The effect of initial pH (4-8), inoculum size (1-20% (v/v)) and glucose concentration (1-30 g L(-1)) on hydrogen production were studied. The optimum cultivation temperature for hydrogen production was at 30 degrees C. The results show that substrate concentration and inoculum size resulted in hydrogen yield (Y(P/S)) of 391 mL g(-1) glucose utilized with maximum hydrogen productivity of 77.5 mL/L/h. Higher substrate concentration or inoculum size adversely affects hydrogen production, which decreases hydrogen yield by 15% to 334 mL g(-1) glucose utilized when 30% (v/v) inoculum size was used. The use of 30 g L(-1) substrate concentration resulted in a 75% decrease to 97 mL g(-1) glucose supplied. Concluded that proper Xo/So enhanced the hydrogen production.

Matched MeSH terms: Clostridium acetobutylicum/metabolism*
Removal of headspace CO2 increases biological hydrogen production by C. acetobutylicum

Alshiyab H, Kalil MS, Hamid AA, Yusoff WM

Pak J Biol Sci, 2008 Oct 01;11(19):2336-40.
PMID: 19137867

The effect of removal of resultant gas resulted in enhancement of the H2 yield. The technique of CO2 scavenging resulted in H2 yield being improved from 408 mL g(-1) to reach the maximum of 422 mL g'. The highest hydrogen productivity of 87.9 ml L(-1) h(-1) was obtained by CO2 scavenging. Biomass concentration was enhanced to 1.47 g L(-1), Y(P,X) of 287 ml g(-1) L(-1), Y(X/S) of 0.294 and Y(H2/s) of 0.0377 by the use of CO2 scavenging. The results suggested that the presence of the gaseous products in fermentation medium and headspace adversely effect biomass growth and hydrogen production.

Matched MeSH terms: Clostridium acetobutylicum/metabolism*
Production of Acetone-Butanol-Ethanol and Hydrogen of Palm Oil Mill Effluent Anaerobic Fermentation with Clostridium Acetobutylicum NCIMB 13357

Pang WK, Wan Mohtar Wan Yusoff, Mohd Sahaid Kalil, Osman Hassan

Sains Malaysiana, 2004;33.

Palm oil mill effluent (POME) can be utilised directly as the sole substrate in the anaerobic fermentation of acetone-butanol-ethanol (ABE) and hydrogen by Clostridium acetobutylicum NClMB13357 in a submerged batch system. Effects of sedimented POME concentration and the initial culture pH on the production of ABE/H were studied. Sedimented POME with 90% v/v (POME90) at pH 5.8 is capable of producing 4.01 g/L ABE with acetone concentration at 1.97 g/L; butanol 1.74 g/L and ethanol 0.3 g/L. The highest concentration of butanol (1.86 g/L) was produced from a culture with initial pH 6.0. The production of hydrogen gas was proportioned to the concentration of POME. The highest hydrogen gas production was at pH 5.5 (31 mL). More than 50% (v/v) of hydrogen gas was produced at different pH except pH 4.5, when only 16% (v/v) or 5 mL of hydrogen was produced.

Matched MeSH terms: Clostridium acetobutylicum
Effect of salts addition on hydrogen production by C. acetobutylicum

Alshiyab H, Kalil MS, Hamid AA, Wan Yusoff WM

Pak J Biol Sci, 2008 Sep 15;11(18):2193-200.
PMID: 19137827

The objective of this study is to investigate the effect of salts addition to fermentation medium on hydrogen production, under anaerobic batch culture system. In this study, batch experiments were conducted to investigate the inhibitory effect of both NaCl and sodium acetate on hydrogen production. The optimum pH and temperature for hydrogen production were at initial pH of 7.0 and 30 degrees C. Enhanced production of hydrogen, using glucose as substrate was achieved. In the absence of Sodium Chloride and Sodium Acetate enhanced hydrogen yield (Y(P/S)) from 350 mL g(-1) glucose utilized to 391 mL g(-1) glucose utilized with maximum hydrogen productivity of 77.5 ml/L/h. Results also show that sodium chloride and sodium acetate in the medium adversely affect growth. Hydrogen yield per biomass (Y(P/X)) of 254 ml/L/g, biomass per substrate utilized (Y(X/S)) of 0.268 and (Y(H2/S) of 0.0349. The results suggested that Sodium at any concentration resulted to inhibit the bacterial productivity of hydrogen.

Matched MeSH terms: Clostridium acetobutylicum/drug effects*; Clostridium acetobutylicum/growth & development; Clostridium acetobutylicum/metabolism*
Fulltext Optimisation of Simultaneous Saccharification and Fermentation (SSF) for Biobutanol Production Using Pretreated Oil Palm Empty Fruit Bunch

Md Razali NAA, Ibrahim MF, Kamal Bahrin E, Abd-Aziz S

Molecules, 2018 Aug 03;23(8).
PMID: 30081514 DOI: 10.3390/molecules23081944

This study was conducted in order to optimise simultaneous saccharification and fermentation (SSF) for biobutanol production from a pretreated oil palm empty fruit bunch (OPEFB) by Clostridium acetobutylicum ATCC 824. Temperature, initial pH, cellulase loading and substrate concentration were screened using one factor at a time (OFAT) and further statistically optimised by central composite design (CCD) using the response surface methodology (RSM) approach. Approximately 2.47 g/L of biobutanol concentration and 0.10 g/g of biobutanol yield were obtained after being screened through OFAT with 29.55% increment (1.42 fold). The optimised conditions for SSF after CCD were: temperature of 35 °C, initial pH of 5.5, cellulase loading of 15 FPU/g-substrate and substrate concentration of 5% (w/v). This optimisation study resulted in 55.95% increment (2.14 fold) of biobutanol concentration equivalent to 3.97 g/L and biobutanol yield of 0.16 g/g. The model and optimisation design obtained from this study are important for further improvement of biobutanol production, especially in consolidated bioprocessing technology.

Matched MeSH terms: Clostridium acetobutylicum
Fulltext Impact of pH and butyric acid on butanol production during batch fermentation using a new local isolate of Clostridium acetobutylicum YM1

Al-Shorgani NKN, Kalil MS, Yusoff WMW, Hamid AA

Saudi J Biol Sci, 2018 Feb;25(2):339-348.
PMID: 29472788 DOI: 10.1016/j.sjbs.2017.03.020

The effect of pH and butyric acid supplementation on the production of butanol by a new local isolate of Clostridium acetobutylicum YM1 during batch culture fermentation was investigated. The results showed that pH had a significant effect on bacterial growth and butanol yield and productivity. The optimal initial pH that maximized butanol production was pH 6.0 ± 0.2. Controlled pH was found to be unsuitable for butanol production in strain YM1, while the uncontrolled pH condition with an initial pH of 6.0 ± 0.2 was suitable for bacterial growth, butanol yield and productivity. The maximum butanol concentration of 13.5 ± 1.42 g/L was obtained from cultures grown under the uncontrolled pH condition, resulting in a butanol yield (YP/
S
) and productivity of 0.27 g/g and 0.188 g/L h, respectively. Supplementation of the pH-controlled cultures with 4.0 g/L butyric acid did not improve butanol production; however, supplementation of the uncontrolled pH cultures resulted in high butanol concentrations, yield and productivity (16.50 ± 0.8 g/L, 0.345 g/g and 0.163 g/L h, respectively). pH influenced the activity of NADH-dependent butanol dehydrogenase, with the highest activity obtained under the uncontrolled pH condition. This study revealed that pH is a very important factor in butanol fermentation by C. acetobutylicum YM1.

Matched MeSH terms: Clostridium acetobutylicum
Fulltext Continuous Butanol Fermentation of Dilute Acid-Pretreated De-oiled Rice Bran by Clostridium acetobutylicum YM1

Al-Shorgani NKN, Al-Tabib AI, Kadier A, Zanil MF, Lee KM, Kalil MS

Sci Rep, 2019 03 15;9(1):4622.
PMID: 30874578 DOI: 10.1038/s41598-019-40840-y

Continuous fermentation of dilute acid-pretreated de-oiled rice bran (DRB) to butanol by the Clostridium acetobutylicum YM1 strain was investigated. Pretreatment of DRB with dilute sulfuric acid (1%) resulted in the production of 42.12 g/L total sugars, including 25.57 g/L glucose, 15.1 g/L xylose and 1.46 g/L cellobiose. Pretreated-DRB (SADRB) was used as a fermentation medium at various dilution rates, and a dilution rate of 0.02 h-1 was optimal for solvent production, in which 11.18 g/L of total solvent was produced (acetone 4.37 g/L, butanol 5.89 g/L and ethanol 0.92 g/L). Detoxification of SADRB with activated charcoal resulted in the high removal of fermentation inhibitory compounds. Fermentation of detoxified-SADRB in continuous fermentation with a dilution rate of 0.02 h-1 achieved higher concentrations of solvent (12.42 g/L) and butanol (6.87 g/L), respectively, with a solvent productivity of 0.248 g/L.h. This study showed that the solvent concentration and productivity in continuous fermentation from SADRB was higher than that obtained from batch culture fermentation. This study also provides an economic assessment for butanol production in continuous fermentation process from DRB to validate the commercial viability of this process.

Matched MeSH terms: Clostridium acetobutylicum/metabolism*
Improvement of the butanol production selectivity and butanol to acetone ratio (B:A) by addition of electron carriers in the batch culture of a new local isolate of Clostridium acetobutylicum YM1

Nasser Al-Shorgani NK, Kalil MS, Wan Yusoff WM, Shukor H, Hamid AA

Anaerobe, 2015 Dec;36:65-72.
PMID: 26439644 DOI: 10.1016/j.anaerobe.2015.09.008

Improvement in the butanol production selectivity or enhanced butanol:acetone ratio (B:A) is desirable in acetone-butanol-ethanol (ABE) fermentation by Clostridium strains. In this study, artificial electron carriers were added to the fermentation medium of a new isolate of Clostridium acetobutylicum YM1 in order to improve the butanol yield and B:A ratio. The results revealed that medium supplementation with electron carriers changed the metabolism flux of electron and carbon in ABE fermentation by YM1. A decrease in acetone production, which subsequently improved the B:A ratio, was observed. Further improvement in the butanol production and B:A ratios were obtained when the fermentation medium was supplemented with butyric acid. The maximum butanol production (18.20 ± 1.38 g/L) was gained when a combination of methyl red and butyric acid was added. Although the addition of benzyl viologen (0.1 mM) and butyric acid resulted in high a B:A ratio of 16:1 (800% increment compared with the conventional 2:1 ratio), the addition of benzyl viologen to the culture after 4 h resulted in the production of 18.05 g/L butanol. Manipulating the metabolic flux to butanol through the addition of electron carriers could become an alternative strategy to achieve higher butanol productivity and improve the B:A ratio.

Matched MeSH terms: Clostridium acetobutylicum
Fulltext Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production

Rahnama N, Foo HL, Abdul Rahman NA, Ariff A, Md Shah UK

BMC Biotechnol, 2014;14:103.
PMID: 25496491 DOI: 10.1186/s12896-014-0103-y

BACKGROUND: Rice straw has shown to be a promising agricultural by-product in the bioconversion of biomass to value-added products. Hydrolysis of cellulose, a main constituent of lignocellulosic biomass, is a requirement for fermentable sugar production and its subsequent bioconversion to biofuels such as biobutanol. The high cost of commercial enzymes is a major impediment to the industrial application of cellulases. Therefore, the use of local microbial enzymes has been suggested. Trichoderma harzianum strains are potential CMCase and β-glucosidase producers. However, few researches have been reported on cellulase production by T. harzianum and the subsequent use of the crude cellulase for cellulose enzymatic hydrolysis. For cellulose hydrolysis to be efficiently performed, the presence of the whole set of cellulase components including exoglucanase, endoglucanase, and β-glucosidase at a considerable concentration is required. Biomass recalcitrance is also a bottleneck in the bioconversion of agricultural residues to value-added products. An effective pretreatment could be of central significance in the bioconversion of biomass to biofuels.
RESULTS: Rice straw pretreated using various concentrations of NaOH was subjected to enzymatic hydrolysis. The saccharification of rice straw pretreated with 2% (w/v) NaOH using crude cellulase from local T. harzianum SNRS3 resulted in the production of 29.87 g/L reducing sugar and a yield of 0.6 g/g substrate. The use of rice straw hydrolysate as carbon source for biobutanol fermentation by Clostridium acetobutylicum ATCC 824 resulted in an ABE yield, ABE productivity, and biobutanol yield of 0.27 g/g glucose, 0.04 g/L/h and 0.16 g/g glucose, respectively. As a potential β-glucosidase producer, T. harzianum SNRS3 used in this study was able to produce β-glucosidase at the activity of 173.71 U/g substrate. However, for cellulose hydrolysis to be efficient, Filter Paper Activity at a considerable concentration is also required to initiate the hydrolytic reaction. According to the results of our study, FPase is a major component of cellulose hydrolytic enzyme complex system and the reducing sugar rate-limiting enzyme.
CONCLUSION: Our study revealed that rice straw hydrolysate served as a potential substrate for biobutanol production and FPase is a rate-limiting enzyme in saccharification.

Matched MeSH terms: Clostridium acetobutylicum/metabolism*