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Article

Biohydrogen Gas/Acetone-Butanol-Ethanol Production from Agave Guishe Juice as a Low-Cost Growing Medium

by
Alejandra G. Oliva-Rodríguez
1,
Vianey de J. Cervantes-Güicho
1,
Thelma K. Morales-Martínez
1,*,
José A. Rodríguez-De la Garza
1,
Miguel A. Medina-Morales
1,
Silvia Y. Martínez-Amador
2,
Ana G. Reyes
3 and
Leopoldo J. Ríos-González
1,*
1
Departamento de Biotecnología, Facultad de Ciencias Químicas, Universidad Autónoma de Coahuila, Boulevard Venustiano Carranza República Oriente, Saltillo 25280, Mexico
2
Departamento de Botánica, Universidad Autónoma Agraria Antonio Narro, Calzada Antonio Narro 1923, Buenavista, Saltillo 25315, Mexico
3
Centro de Investigaciones Biológicas del Noroeste, La Paz 23096, Mexico
*
Authors to whom correspondence should be addressed.
Fermentation 2023, 9(9), 811; https://doi.org/10.3390/fermentation9090811
Submission received: 28 July 2023 / Revised: 8 August 2023 / Accepted: 11 August 2023 / Published: 5 September 2023
(This article belongs to the Section Fermentation Process Design)
Browse Figures
Review Reports Versions Notes

Abstract

:
Different strategies have been assessed for the revalorization of guishe to obtain biomolecules. The juice obtained after the mechanical extraction of guishe is rich in phytochemicals and sugars which can be converted to other products. The objective of the present study was to evaluate the production of hydrogen and butanol at different guishe juice concentrations (and therefore, different sugar concentrations) via fermentation in batch mode using Clostridium acetobutylicum ATCC 824. Fermentation assays were performed in triplicate under anaerobic conditions at 35 °C for 142 h. Guishe juice was supplemented with all components of synthetic medium (salts, vitamins and reducing agents), except glucose, and diluted at different concentrations: 20%, 40%, 60%, 80% and 100%. For comparison purposes, a control was carried out in a synthetic medium using glucose as carbon source. Results showed a maximum butanol concentration of 5.39 g/L using 80% guishe juice, corresponding to a productivity and yield of 0.04 g/L h−1 and 0.24 g/g, respectively. Meanwhile, the highest productivity (1.16 L H2/L d−1; 1.99 mmol H2/L h−1) and yield (18.4 L/kg) of hydrogen were obtained with 40% guishe juice. This study demonstrates the potential of guishe juice to be used as a low-cost substrate for hydrogen and butanol production.

1. Introduction

The growing energy demand and environmental effects due to society’s dependence on fossil fuels have encouraged the search for renewable energy supplies. Among many alternatives, biofuels produced from renewable feedstocks have gained attention from researchers worldwide [1,2].
Out of the various biomass-based fuels, hydrogen is an ideal alternative, mainly because of two significant advantages: zero carbon emissions, since the only by-product is water vapor, and higher energy content (122 kJ·g−1) compared to other fuels, as it produces almost three times more energy than gasoline [3,4]. Butanol is also a promising biofuel due to its many similarities to gasoline and its superior characteristics over other biofuels, such as lower volatility and corrosivity, higher viscosity and greater energy content. These distinctive characteristics allow butanol to be blended in any proportion with gasoline and diesel or used as a complete replacement for these fossil fuels without modifying the current engine systems or fuel distribution infrastructure [2,5,6].
Clostridium species are Gram-positive and strict anaerobic bacteria and can produce butanol and hydrogen via acetone–butanol–ethanol (ABE) fermentation. ABE fermentation is a well-known bioprocess for butanol production from fermentable sugars. It has been used since the beginning of the 20th century and has gained scientific attention due to increased interest in renewable fuels [7,8]. In recent years, the genus Clostridium has been extensively studied for fermentative hydrogen production, as it has important advantages such as high hydrogen yield, a wide range of substrates and resistance to adverse conditions [9].
However, ABE fermentation has several challenges, with cost feasibility being one of the most relevant since the cost of raw materials to produce first-generation butanol and hydrogen represents 70% of the production costs [10]; therefore, selecting a suitable substrate is crucial. In this context, considering agricultural wastes with high content of fermentable sugars as low-cost feedstock offers important benefits, not only in terms of production costs, but also in terms of waste management.
In recent studies, Clostridium acetobutylicum was able to produce butanol from Agave lechuguilla biomass (cogollo) hydrolysates [11] and to co-produce hydrogen using a mixed culture [12]. Although producing these fuels from cellulose is an alternative in conceptualizing guishe biorefinery, there are still technical–economic aspects to overcome.
Mexico is one of the main producers of agroindustrial waste worldwide, which could be used to obtain high-value products [13]. Particularly, rural communities in arid zones of northeastern Mexico generate at least 150,000 tons of waste every year due to fiber extraction from Agave lechuguilla, which is their main economic activity [14]. This fiber, known as ixtle, is used to fabricate products such as brushes, ropes, carpets and handbags, among many other items [15]. The amount of waste from this activity is so high because ixtle represents only 15% of the plant; therefore, around 850 kg of guishe (term used to describe bagasse exclusively from Agave lechuguilla) are generated per ton of processed (carved) leaves of Agave lechuguilla [16]. Currently, producers burn or accumulate discard opencast this waste, causing environmental damage to soil, water and air, and even causing death to farm animals when ingested due to its high fiber content [15,17].
Different strategies have been studied for the revalorization of guishe to obtain high-value compounds, mainly for extracting flavonoids and saponins [18,19]. Recently, Díaz-Jiménez et al. [15] proposed the first concept of a guishe biorefinery through mass balance analysis, considering the production of ethanol, biogas, syngas and biochar from the solid fraction of filtered guishe. Currently, only the production of syngas and biochar has been evaluated from guishe [15], while ethanol, butanol and hydrogen have been extensively studied using the biomass of the soft central leaves known as “cogollo” without carving [11,12,20,21,22,23,24].
Guishe can be separated into two fractions by mechanical pressing. The liquid fraction, a dark green extract (that will be further referred to as guishe juice or Agave lechuguilla juice) [4], has not yet been studied for the production of biofuels. In addition to the phytochemical content, guishe juice is rich in sugars, which can be converted to high-value products such as hydrogen, and other biofuels like ethanol or butanol. The antimicrobial activity of polyphenols and saponins is well known; however, the effect varies from one microorganism to another. Consequently, regardless of the product of interest, ensuring that the microorganisms in the process grow optimally in the presence of these phytochemicals is one of the main challenges to overcome [25,26].
Considering the amount of guishe generated every year that can be used as a low-cost substrate for the ABE fermentation process without being submitted to any previous process, the aim of the present work was to evaluate hydrogen and ABE (acetone, butanol and ethanol) production from the liquid fraction of guishe using Clostridium acetobutylicum ATCC 824 as inoculum under anaerobic conditions in batch reactors.

2. Materials and Methods

2.1. Raw Material Collection

Agave lechuguilla cogollos were harvested and processed by peasants from Ejido Cosme, in the municipality of Ramos Arizpe, Coahuila, Mexico (GPS: 25°52′3.6″ N; 101°19′51.1″ W), according to Mexican regulations for its exploitation [16]. The guishe generated during this process was collected and subjected to mechanical pressing to obtain two fractions. The solid fraction was not used in this study. The liquid fraction or guishe juice was subsequently bottled in plastic containers and stored in the dark at −20 °C until further use for experimental assays.

2.2. Guishe Juice Characterization

Sugar content of guishe juice was determined by high performance liquid chromatography (HPLC) [12]. Total polyphenolic content (TPC) and total flavonoid content (TFC) were quantified as described by Morreeuw et al. [14]. The biochemical oxygen demand (BOD) and chemical oxygen demand (COD) were determined according to Pérez-Rodríguez et al. [27]. Determination of ashes, expressed as the percentage of residue remaining after dry oxidation at 550 to 600 °C, was carried out using the analytical method NREL/TP-510-42622 [28]. For elemental analysis, guishe juice was first dried in a dehydrator (Koleff KL-10, México), then 1 g of dried sample (after drying, the samples turned into a fine powder) was analyzed by X-ray fluorescence spectroscopy using an Epsilon 1 XRF Spectrometer from Malvern Panalytical (Enigma Business Park, UK) and equipped with Omnian software.

2.3. Microorganisms

Clostridium acetobutylicum ATCC 824 was propagated under anaerobic conditions in 120 mL glass bottles containing 70 mL of a synthetic medium at pH 6.7 and 35 °C as described by Oliva-Rodríguez et al. [11] for 72 h. The synthetic medium was formulated as described by Medina-Morales et al. [29] using glucose as carbon source at a concentration of 60 g/L.

2.4. Experimental Setup

The effect of different guishe juice concentrations (and therefore, different sugar content) on hydrogen and ABE production was evaluated: 20%, 40%, 60%, 80% and 100%. Guishe juice was supplemented with all components of synthetic medium (salts and vitamins), except glucose, adjusting the pH to 6.7. A control was carried out in a synthetic medium with glucose as carbon source (0% guishe juice) at a concentration of 60 g/L.
After adding the medium, the reactors were sealed with rubber stoppers and aluminum caps. Once sealed, the reactors were purged with nitrogen to remove oxygen in the gaseous phase and generate anaerobic conditions. All reactors were sterilized and then inoculated with 6.5 mL f C. acetobutylicum cells.
Fermentation assays were performed under anaerobic conditions at 35 °C for 142 h in 120 mL glass bottles with 65 mL of working volume, and liquid and gas samples were taken every 24 h. Gas production was measured by displacement using a 5% NaOH solution, and the presence of hydrogen in gaseous samples was determined by gas chromatography-TCD. Liquid samples were analyzed to quantify sugar content by high performance liquid chromatography (HPLC) and butanol by gas chromatography-FID.

2.5. Analytical Methods

ABE production was determined by GC-FID (Agilent 7890, Santa Clara, CA, USA) equipped with BR-Swax column (30 m × 0.32 mm ID, film thickness 0.25 μm), using nitrogen gas as a carrier, with an initial column temperature of 59 °C and a final temperature of 190 °C for a total run time of 15 min; detector temperature of 300 °C and an injector temperature of 250 °C. To confirm hydrogen production, a gaseous sample of 25 µL from the reactors was injected into a GC TCD (Varian 3400, Santa Clara, CA, USA) equipped with a CP-Molsieve 5A column (30 m × 0.53 mm ID, film thickness 0.15 μm) using argon gas as carrier, with a column temperature of 50° C, and detector and injector temperature of 200 °C, with a total run time of 4 min.
Sugar content (glucose and xylose) was analyzed by HPLC in an Agilent 1260 Infinity liquid chromatograph (CA, USA) equipped with a refraction index detector at 45 °C and an Agilent Hi-Plex H column at 35 °C using 5 mM H2SO4 solution as mobile phase (0.5 mL/min) [12].

2.6. Data Analyses

All experiments were carried out in triplicate, and results were analyzed in Microsoft Excel for means, standard deviations, and graphics. Analysis of variance and comparison of means by Tukey’s test (at 0.05) using Minitab® 21.1 statistical software was used for butanol and hydrogen experiment results.

3. Results

3.1. Guishe Juice Characterization

Guishe juice characterization results are shown in Table 1. BOD corresponds to the amount of potentially fermentable material (>90% of organic matter compared to the COD). The sum of glucose and xylose in guishe juice resulted in a concentration of 35 g/L. For a total polyphenolic content (TPC) that was equivalent to 11.4 gallic acid milligrams (GAE) and total flavonoid content (TFC) that was equivalent to 7.6 quercetin milligrams (QE) per gram of guishe juice, respectively. Finally, ash content that corresponded to 12%, was mainly composed of Ca, K and Mg; these minerals could represent at least part of the soil composition where the Agave lechuguilla plant grew [12].

3.2. Butanol and Hydrogen Production

Figure 1 shows the butanol production from agave juice at different sugar concentrations compared to the control prepared with commercial glucose. The maximum butanol production (5.4 g/L at 142 h) was obtained with 80% agave juice, which is 32% less than the amount obtained in synthetic medium (control).
Butanol production performance can be divided into two sections: (1) at low concentrations of guishe juice (20%, 40% and 60%), butanol production was faster and more than 70% was produced in the first 70 h; (2) at high concentrations (80% and 100%), the start was slow, but it increased significantly from 47 h to surpass the amount obtained in experiments at low concentrations. Fermentation with guishe juice undiluted (100%) produced slightly less butanol (4.9 g/L) than the experiment at 80%.
Table 2 shows the profile of ABE solvents produced, along with butanol productivity and yield, under the conditions studied. Productivity was calculated as the maximum butanol concentration, divided by the time necessary to reach that concentration. In all cases, the production of ethanol and acetone was low compared to the theoretical value (3:6:1—acetone:butanol:ethanol, respectively). The sum of solvents also resulted in a maximum value using 80% guishe juice (7.09 g/L) compared with experiments at 20% (4.57 g/L). However, it can be observed that diluting the guishe juice to 40% resulted in higher productivity (52.1 mg/L h−1), and diluting it more, up to 20%, improved production yield with respect to sugar consumption (0.43 g/g sugars consumed).
As expected, the slow butanol production at 80% and 100% of guishe juice concentrations negatively affected the productivity. In comparison, the difference between the production of butanol and the low consumption of sugars at 20% of guishe juice could promote a high yield.
Overall, butanol concentration was favorable at 80% guishe juice; maximum productivity was obtained at 40% guishe juice, and the highest yield was obtained at 20% guishe juice. The advantages of high butanol concentrations during the separation process are discussed below.
Hydrogen accumulation during fermentation performed similarly to butanol production (Figure 2). Although hydrogen production in the control was constant during the 142 h of fermentation, the maximum production was obtained using 80% guishe juice (351 mL). Experiments at high concentrations of guishe juice (80% and 100%) also showed a delay in hydrogen production during the first 47 h.
Comparing both processes shows that hydrogen production stopped; meanwhile, butanol was continuously produced until the end of fermentation. Although at 20% guishe juice, production was lower (149 mL), this was obtained in only 47 h, followed by concentrations of 40% (226 mL at 70 h) and 60% (276 mL at 94 h). This rapidity translated into a high productivity (Table 3), reaching a maximum at 40% guishe juice (1.1 L H2/L d−1 and 1.99 mmol/L h−1).
Conversely, productivity was negatively affected in slow experiments (80%, 100% and control). Also, hydrogen yield with respect to sugar consumption was highest at 40% guishe juice (18.4 mL/g sugars consumed); therefore, all fermentation parameters shown in Table 3 were higher at this concentration.
Figure 3 shows the sugar consumption at different guishe juice concentrations compared to the control. Comparing Figure 1, Figure 2 and Figure 3, it can be observed that the production of butanol and hydrogen is directly related to the consumption of sugars. The performance in the consumption of sugars in the lowest concentrations of guishe juice (20%, 40% and 60%) was similar to the control during the first 70 h. Meanwhile, at 80% and 100% guishe juice, sugar consumption started after 47 h, reaching a final concentration of 6.6 g/L and 14.3 g/L, respectively.
In all experiments, sugar consumption yield was higher using guishe juice compared to the control (Table 4). Sugar consumption increased from 9.4 g/L to 22.6 g/L at guishe juice concentrations from 20% to 80%, respectively, then decreased at 100% (20.6 g/L, corresponding to a yield of 59.1%).
When calculating the sugar consumption yield, an inverse effect is observed due to the difference with respect to the initial concentration. Therefore, the maximum sugar consumption yield was obtained at 20% of guishe juice (81.1%), followed by the experiment at 40% (79.2%).

4. Discussion

4.1. Guishe Juice Characterization

According to previous results, the ash content in guishe oscillates between 4 and 7% [15]; therefore, the increase in the ash content can be attributed to a higher amount of inorganic compounds that are concentrated during the mechanical pressing of the guishe. The TPC and TFC content detected in the guishe juice are similar to those reported by Morreeuw et al. [14] (10.46 mg GAE/g and 4.53 mg QE/g, respectively) in guishe from Agave lechuguilla collected in the same location. These slight differences are mainly due to the temporality of the collected samples and the pretreatment process employed. Recently, Sánchez et al. [30] reported a total sugar concentration for guishe juice of 23.3 g/L, less than the result obtained in the present work (35 g/L).
Although high sugar content is desirable for ABE fermentation, it has been reported that solventogenic microorganisms such as C. acetobutylicum do not consume the total substrate present, consuming around 40–50% in synthetic medium and 30–35% in hydrolysates from different sources [11,31,32]. Therefore, it was expected that sugar content in the guishe juice used in this study would be enough for adequate production of hydrogen and butanol by fermentation.

4.2. Butanol and Hydrogen Production

Ideal guishe juice concentration for butanol production will depend on the final butanol concentration in the fermentation broth, productivity and yield. However, butanol concentration influences the energy requirements and the amount of wastewater generated during the distillation–separation process. In other words, the viability of the process depends largely on the butanol titers obtained in the fermentation process [33].
It has been estimated that the cost of butanol separation obtained from an ABE fermentation process represents about 14% of the cost of production [34]. According to Chen et al. [35], ~79 MJ/kg would be required to separate butanol from a binary butanol–water mixture at 0.5% (w/v) by a traditional distillation system (maximum butanol concentration obtained at 80% guishe juice). This amount of energy required exceeds the amount obtained from butanol (36 MJ/kg); however, if fermentation could result in a butanol concentration of 40 g/L (4% w/v), the energy required would significantly decrease.
ABE fermentation processes are still not economically viable due to low butanol titers (~20 g/L) [36]. Therefore, searching for cheap feedstocks and genetic improvement in strains is imperative to reduce production costs. Table 5 shows that despite this being an initial assessment of the use of guishe juice as a low-cost substrate, butanol concentration, yield and productivity are competitive compared to other works. Another favorable aspect of the results obtained in the present work is that in most of the reports that are compared, the substrates correspond to lignocellulosic and algal biomass, which normally require several stages prior to fermentation.
Compared to previous work by our research group [11], the maximum butanol production in the experiment at 80% guishe juice is similar to that obtained from enzymatic hydrolysates of Agave lechuguilla central soft leaves (guishe + fiber) pretreated by autohydrolysis (6.1 g/L at 96 h of fermentation). However, for production from agave cellulose, biomass must be dried, milled, pretreated (at temperatures between 180 °C and 190 °C) and hydrolyzed using commercial enzymatic complexes before carrying out the ABE fermentation process.
Unlike butanol production, fermentation parameters obtained during hydrogen generation are competitive compared to other reports. Hydrogen productivity obtained at 40% guishe juice (1.99 mmol H2/L h−1) is higher than the productivity reported by Li et al. [43] obtained from sugarcane juice by Ethanoligenens harbinense in a single batch mode (1.73 mmol/L h−1).
Numerous reports can be found using agave biomasses (mainly from Agave tequilana bagasse) as feedstock for hydrogen production in continuous mode. The hydrogen productivity in these reports varies from 0.1 to 13 L H2/L d−1, but with the difference that in most of the reports a pretreatment was required in which the biomass obtained after the pretreatment needs acid or enzymatic hydrolysis, and in some cases, this hydrolysate required further processing for detoxification [44,45,46,47,48,49,50,51,52,53]. Comparing the maximum productivity obtained (1.5 L H2/L d−1) with the only report in batch mode (to our knowledge) that used A. tequilana baggase to produce hydrogen, by Tapia-Rodríguez et al. [53] (0.85 L H2/L d−1), the potential of guishe juice as substrate can be appreciated.
The increased assimilation of sugars in experiments with guishe juice compared to the control can be attributed to the presence of saponins. Glycoside saponins interact with membrane hopanoids favoring the transportation activity by forming reversible pores [54]. The surfactant properties of saponins in aqueous solutions can be attributed to their amphiphilic structure, a combination of lipophilic non-polar aglycone and hydrophilic polar glycone moieties [55]. This structural feature of a saponin molecule resembles that of a synthetic surfactant molecule [56]. Xin et al. [57] reported that the increase in the concentration of tween 80 during butanol production by C. acetobutylicum promoted greater assimilation of sugars.
However, a high concentration of saponins can damage cell membrane integrity. Alcázar-Valle et al. [58] mentioned that the phospholipids of the cell membrane of Kluyveromyces marxianus and Saccharomyces cerevisiae were affected by the reduction and unsaturation of acyl chains when both yeasts interacted with agave saponin extracts. This phenomenon could occur in the case of the concentration of 100% guishe juice, as the production of butanol and hydrogen decreased significantly compared to experiments with diluted guishe juice.

5. Conclusions

The results showed that C. acetobutylicum ATCC 824 could grow and produce butanol and hydrogen via ABE fermentation from the guishe juice of A. lechuguilla as low-cost feedstock. However, the production of both compounds was better when guishe juice was diluted from 40% to 80%. In all cases, the presence of surfactant molecules, such as saponins in guishe juice, could favor the assimilation of sugars, but at 100%, the consumption of sugars and, therefore, the production of butanol and hydrogen were negatively affected.
The selection of guishe juice concentration will depend on the product of interest. In the case of butanol, maximum production was achieved using 80%, while at 40% the best kinetic parameters of hydrogen production were obtained. The results obtained so far suggest that yields can be significantly increased once the process is optimized and assessed in continuous mode in future research. Finally, these early findings can allow further advances in biorefinery conceptualization from the guishe of Agave lechuguilla.

Author Contributions

Conceptualization, L.J.R.-G. and T.K.M.-M.; methodology, A.G.O.-R., V.d.J.C.-G., S.Y.M.-A. and A.G.R.; software, T.K.M.-M.; validation, A.G.O.-R.; formal analysis, T.K.M.-M.; investigation, L.J.R.-G. and A.G.O.-R.; resources, L.J.R.-G.; data curation, A.G.O.-R.; writing—original draft preparation, A.G.O.-R.; writing—review and editing, L.J.R.-G., J.A.R.-D.l.G., T.K.M.-M. and M.A.M.-M.; visualization, J.A.R.-D.l.G.; supervision, L.J.R.-G.; project administration, L.J.R.-G.; funding acquisition, L.J.R.-G. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Consejo Estatal de Ciencia y Tecnología (COECYT) through project number COAH-2022-C19-C071.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Acknowledgments

To Consejo Nacional de Humanidades, Ciencias y Tecnologías (CONAHCyT) by granting scholarships to students.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Butanol production by C. acetobutylicum ATCC 824 at different guishe juice concentrations.
Figure 1. Butanol production by C. acetobutylicum ATCC 824 at different guishe juice concentrations.
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Figure 2. Hydrogen production by C. acetobutylicum ATCC 824 at different guishe juice concentrations compared to the control using commercial glucose.
Figure 2. Hydrogen production by C. acetobutylicum ATCC 824 at different guishe juice concentrations compared to the control using commercial glucose.
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Figure 3. Sugar consumption by C. acetobutylicum ATCC 824 at different guishe juice concentrations compared to the control using commercial glucose.
Figure 3. Sugar consumption by C. acetobutylicum ATCC 824 at different guishe juice concentrations compared to the control using commercial glucose.
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Table 1. Guishe juice characterization.
Table 1. Guishe juice characterization.
ParameterValue
Chemical analysisCOD (g/L)300 ± 7.1
BOD (g/L)276 ± 8.3
* TPC (mg GAE/g)11.4 ± 0.7
* TFC (mg QE/g)7.6 ± 0.4
** Sugar content (g/L)35.0 ± 2.8
Ash (% w/w)12.0 ± 0.9
Elemental analysisCa (% w/w)5.4 ± 0.2
K (% w/w)4.1 ± 0.3
Mg (% w/w)1.9 ± 0.1
* GAE: Gallic acid equivalent; QE: Quercetin equivalent; ** Sum of glucose and xylose. BOD: Biochemical oxygen demand, COD: Chemical oxygen demand, TPC: Total polyphenolic content; TFC: Total flavonoid content
Table 2. ABE production and butanol productivity by C. acetobutylicum ATCC 824 at different guishe juice concentrations.
Table 2. ABE production and butanol productivity by C. acetobutylicum ATCC 824 at different guishe juice concentrations.
Juice Concentration (%)Solvent Concentration (g/L)Total ABE Concentration (g/L)Butanol Productivity (mg/L h−1)Butanol Yield
(g/g Sugars Consumed)
AcetoneButanolEthanol
Control2.83 ± 0.127.97 ± 0.300.17 ± 0.0110.97 ± 0.4390.3 ± 3.70.30 ± 0.01
200.52 ± 0.01 E4.01 ± 0.08 C0.04 ± 0.01 C4.57 ± 0.10 E25.3 ± 1.4 E0.43 ± 0.02 A
400.97 ± 0.08 D3.98 ± 0.26 C0.08 ± 0.01 B5.03 ± 0.35 D52.1 ± 3.2 A0.30 ± 0.01 B
601.26 ± 0.06 C4.66 ± 0.13 B0.12 ± 0.01 A6.04 ± 0.20 C51.2 ± 3.6 B0.30 ± 0.02 B
801.55 ± 0.06 A5.39 ± 0.19 A0.14 ± 0.00 A7.09 ± 0.25 A38.9 ± 1.3 C0.24 ± 0.01 C
1001.42 ± 0.13 B4.96 ± 0.18 B0.14 ± 0.01 A6.52 ± 0.32 B36 ± 2.2 D0.24 ± 0.01 C
Means difference is significant at a level of 0.05. Significantly different means do not share a letter.
Table 3. Hydrogen productivity and yield at different guishe juice concentrations by C. acetobutylicum ATCC 824.
Table 3. Hydrogen productivity and yield at different guishe juice concentrations by C. acetobutylicum ATCC 824.
Juice Concentration (%)Productivity (L H2/L d−1)Productivity (mmol H2/L h−1)Hydrogen Yield (mL/g Sugars Consumed)
Control0.75 ± 0.001.28 ± 0.0110.9 ± 0.13
201.10 ± 0.01 A1.88 ± 0.03 B17.4 ± 0.30 B
401.16 ± 0.02 A1.99 ± 0.04 A18.4 ± 0.39 A
601.08 ± 0.04 A1.86 ± 0.07 B18.6 ± 0.74 A
800.91 ± 0.01 B1.56 ± 0.01 C15.5 ± 0.17 D
1000.86 ± 0.03 B1.48 ± 0.05 D16.1 ± 0.59 C
Means difference is significant at the 0.05 level. Means that do not share a letter are significantly different.
Table 4. Sugar consumption by C. acetobutylicum ATCC824 at different agave juice concentrations.
Table 4. Sugar consumption by C. acetobutylicum ATCC824 at different agave juice concentrations.
Juice Concentration (%)Sugar Consumption (g/L)Sugar Consumption Yield (%)
Control26.2 ± 0.5541.6 ± 2.1
209.4 ± 0.10 E81.1 ± 1.1 A
4013.4 ± 0.33 D79.2 ± 2.5 B
6015.8 ± 0.26 C72.1 ± 1.7 D
8022.6 ± 0.29 A77.3 ± 1.3 C
10020.6 ± 0.37 B59.1 ± 1.8 E
Means difference is significant at the 0.05 level. Means that do not share a letter are significantly different.
Table 5. Comparison of butanol production performance from different substrates.
Table 5. Comparison of butanol production performance from different substrates.
SubstrateMicroorganismButanol (g/L)Butanol Yield
(g/g Sugars Consumed)		Butanol Productivity (g/L h−1)Ref.
Rice straw hydrolysateC. acetobutylicum ATCC 8249.100.170.79[31]
Sugar cane bagasse hydrolysateC. acetobutylicum ATCC 8248.400.160.80[31]
Domestic organic wasteC. acetobutylicum ATCC 8247.80-----0.065[37]
Wood waste (Pinus rigida)C. beijerinckii NCIMB 80526.910.25-----[38]
Rice straw hydrolysateC. saccharoperbutylacetonicum ATCC 135646.680.090.28[39]
Sweet potato vineC. acetobutylicum ATCC 8246.400.180.09[40]
A. lechuguilla hydrolysateC. acetobutylicum ATCC 8246.100.270.073[11]
A. lechuguilla juiceC. acetobutylicum ATCC 8245.390.240.04This study
Pineapple waste hydrolysateC. acetobutylicum B 5175.230.150.05[41]
Microalgae biomassC. acetobutylicum ATCC 8244.36----------[31]
Oleaginous fungal biomass (Cunninghamella echinulate)C. acetobutylicum ATCC 8242.190.450.072[42]
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Oliva-Rodríguez, A.G.; Cervantes-Güicho, V.d.J.; Morales-Martínez, T.K.; Rodríguez-De la Garza, J.A.; Medina-Morales, M.A.; Martínez-Amador, S.Y.; Reyes, A.G.; Ríos-González, L.J. Biohydrogen Gas/Acetone-Butanol-Ethanol Production from Agave Guishe Juice as a Low-Cost Growing Medium. Fermentation 2023, 9, 811. https://doi.org/10.3390/fermentation9090811

AMA Style

Oliva-Rodríguez AG, Cervantes-Güicho VdJ, Morales-Martínez TK, Rodríguez-De la Garza JA, Medina-Morales MA, Martínez-Amador SY, Reyes AG, Ríos-González LJ. Biohydrogen Gas/Acetone-Butanol-Ethanol Production from Agave Guishe Juice as a Low-Cost Growing Medium. Fermentation. 2023; 9(9):811. https://doi.org/10.3390/fermentation9090811

Chicago/Turabian Style

Oliva-Rodríguez, Alejandra G., Vianey de J. Cervantes-Güicho, Thelma K. Morales-Martínez, José A. Rodríguez-De la Garza, Miguel A. Medina-Morales, Silvia Y. Martínez-Amador, Ana G. Reyes, and Leopoldo J. Ríos-González. 2023. "Biohydrogen Gas/Acetone-Butanol-Ethanol Production from Agave Guishe Juice as a Low-Cost Growing Medium" Fermentation 9, no. 9: 811. https://doi.org/10.3390/fermentation9090811

APA Style

Oliva-Rodríguez, A. G., Cervantes-Güicho, V. d. J., Morales-Martínez, T. K., Rodríguez-De la Garza, J. A., Medina-Morales, M. A., Martínez-Amador, S. Y., Reyes, A. G., & Ríos-González, L. J. (2023). Biohydrogen Gas/Acetone-Butanol-Ethanol Production from Agave Guishe Juice as a Low-Cost Growing Medium. Fermentation, 9(9), 811. https://doi.org/10.3390/fermentation9090811

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