• International Journal of Technology (IJTech)
  • Vol 12, No 2 (2021)

Exploring Starch Sources for the Refreshment Process of Acetone-Butanol-Ethanol Fermentation with Clostridium Saccharoperbutylacetonicum N1-4

Exploring Starch Sources for the Refreshment Process of Acetone-Butanol-Ethanol Fermentation with Clostridium Saccharoperbutylacetonicum N1-4

Title: Exploring Starch Sources for the Refreshment Process of Acetone-Butanol-Ethanol Fermentation with Clostridium Saccharoperbutylacetonicum N1-4
Rizki Fitria Darmayanti, Ari Susanti, Felix Arie Setiawan, Meta Fitri Rizkiana, Maktum Muharja, Bimo Bayu Aji, Mizanurafi' Ghifarhadi Prasiefa, Liony Trisinta Dewi, Zanuba Anggie Yanti

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Cite this article as:
Darmayanti, R.F., Susanti, A., Setiawan, F.A., Rizkiana, M.F., Muharja, M., Aji, B.B., Prasiefa, M.G., Dewi, L.T., Yanti, Z.A., 2020. Exploring Starch Sources for the Refreshment Process of Acetone-Butanol-Ethanol Fermentation with Clostridium Saccharoperbutylacetonicum N1-4. International Journal of Technology. Volume 12(2), pp. 309-319

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Rizki Fitria Darmayanti Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Ari Susanti Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Felix Arie Setiawan 1. Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia 2. Chemical and Biological Engineering Department, Un
Meta Fitri Rizkiana Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Maktum Muharja Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Bimo Bayu Aji Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Mizanurafi' Ghifarhadi Prasiefa Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Liony Trisinta Dewi Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Zanuba Anggie Yanti Department of Chemical Engineering, Faculty of Engineering, University of Jember, Jalan Kalimantan No. 37, Tegal Boto, Jember 68121, Indonesia
Email to Corresponding Author

Abstract
Exploring Starch Sources for the Refreshment Process of Acetone-Butanol-Ethanol Fermentation with Clostridium Saccharoperbutylacetonicum N1-4

Biobutanol is a renewable fuel that can be used as a gasoline substitute and a chemical feedstock. Its production using the Clostridial bacterial strain involves three steps: refreshment from a stock, a preculture for bacterial propagation, and primary fermentation for butanol production. Refreshment is an important process to activate the bacteria and multiply the stock. This process uses potato glucose media for C. saccharoperbutylacetonicum N1-4, while the use of starch from other sources has not been studied. This study aimed to understand various carbon sources’ effects on this refreshment process as part of ABE (acetone-butanol-ethanol) fermentation. Starch was substituted in refreshment media with several types of potato, rice, sweet corn, and sweet potato at 15% w/v. After 24 hours of refreshment at an ambient temperature, fermentation was run for 48 hours in TYA (tryptone-yeast-acetate) glucose media. All the starch sources could be used in the refreshment process, resulting in butanol and total solvent concentration ranging from 7.58 to 8.76 g/L and 12.5 to 14.6 g/L, respectively. Among the samples, sweet corn provided the highest fermentation performance, with butanol of 8.76 g/L, total solvents of 14.6 g/L, average butanol productivity of 0.182 g/L/h, and a butanol yield per substrate of 0.481 C-mol/C-mol. All the starchy materials used in this experiment offered potential for ABE fermentation, while sweet corn performed remarkably—producing the highest final butanol concentration, productivity, and yield.

Biobutanol; Potato; Rice; Refreshment; Sweet corn; Sweet potato

Introduction

The growing population’s increasing energy demand has led to an urgent quest for new energy sources (Yuliansyah et al., 2019). Transportation is one of the highest energy-consuming sectors, requiring specific fuel properties (Febrianti et al., 2017). Biobutanol is among the biofuels that can be used as a substitute for gasoline in premixed combustion engines (Szulczyk, 2010). It offers better properties than ethanol vis-à-vis higher energy value, lower vapor pressure, and an octane number more similar to gasoline. No modification is needed to combust butanol in current engines, and butanol’s application has improved engine performance (Merola et al., 2012; Lapuerta et al., 2017).

Butanol has been produced using biomass feedstock by converting carbohydrates. Sugar and other carbohydrates are digestible using Clostridial species via acidogenesis and solventogenesis phases (Tashiro et al., 2013). One species can produce a large amount of butanol at an ambient temperature: Clostridium saccharoperbutylacetonicum. This strain can directly ferment various types of sugar and starch substrates, with or without hydrolysis (Zhao et al., 2018; Darmayanti et al., 2019).

Starchy vegetables have been produced at large scales to fulfill food needs (Supramono et al., 2016). Rice, potato, corn, and sweet potato are grown widely around the world in increasing quantities and qualities (Devaux et al., 2014; Jusuf and Ginting, 2014; Muthayya et al., 2014). Starchy materials derived directly from fresh vegetables provide various nutrients, especially starch, sugars, protein, and such elements as nitrogen, potassium, magnesium, sulfur, and calcium (McGill et al., 2013). Vitamins are also present in these materials, such as vitamin B in rice (Liu et al., 2019), vitamins C and E in corn, and vitamins A and C in potato and sweet potato (McGill et al., 2013). These nutrients have helped the ABE fermentation strain grow and achieve better viability (Ambarsari and Sonomoto, 2012; Mukherjee et al., 2019).

Culturing C. saccharoperbutylacetonicum from stock generally involves three main steps: refreshment of the stock with heat-shocking, a preculture to grow more cells for larger-scale fermentation, and the main fermentation in a large container for butanol production. The refreshment step has commonly used potato glucose media, which is easily made majorly from potato (15%), glucose, ammonium sulfate, and calcium carbonate (Darmayanti et al., 2018; Hastuti et al., 2019; Zhao et al., 2019). This strain differs considerably from other Clostridial strains used as commercial media, such as reinforced clostridial media (RCM), cooked meat medium (CMM), or clostridial growth medium (CGM), which contain protein and amino acids as their main components (Li et al., 2011; Xue et al., 2012; Qureshi et al., 2014). Refreshment is a critical step to activate inactive bacteria in stock, and this step is also necessary for the stock multiplication process.

To our knowledge, the starchy materials used to refresh C. saccharoperbutylacetonicum have only included potatoes, especially the variety of May queen variety—large-sized potatoes that are grown mostly in Japan and Europe. Among Japan’s potato varieties, only in may-queen potato media allows this strain to grow; it has been unable to grow using other varieties (Gao et al., 2016). Other starchy materials offer potential for use as the refreshment process’s carbon source. While the starch content of potato, rice, corn, and sweet potato is over 30%, their sugar content differs considerably, and their other components might affect cell growth and fermentation (Ambarsari and Sonomoto, 2015). ABE fermentation using different carbon sources for refreshment has not been studied. While vitamins and minerals have been reported to enhance biobutanol production (Li et al., 2014), most are present in the starch sources with various compositions; therefore, ABE fermentation using these refreshment substrates required investigation.

Accordingly, this study aimed to understand the potency and effects of various carbon sources in the refreshment process of ABE fermentation using C. saccharoperbutylacetonicum N1-4. The starch in refreshment media was substituted with several types of potato, rice, sweet corn, and sweet potato. We expected this substitution to widen the variety of starch sources, consequently expanding supplies for these sources. Cell growth, substrate consumption, and solvent production were studied to observe fermentation performance. These experiments revealed sweet corn as a potential starchy vegetable for biobutanol-producing refreshment media. Sweet corn was observed to result in the highest butanol concentration as a refreshment substrate.

Conclusion

We studied starchy materials’ potency for the refreshment process of C. saccharoperbutylacetonicum N1-4 fermentation, using materials that were extensively produced around the world. Potato, rice, sweetcorn, and sweet potato resulted in remarkable butanol concentrations, ranging from 7.58 to 8.76 g/L. Compared with the previous literature, our study’s fermentation using sweet corn as a refreshment media yielded total solvents of 0.496 g/g. Our use of sweet corn as a refreshment substrate resulted in our study’s highest butanol concentration, productivity, and yield because of the supporting minerals present in sweet corn. A further study is required to apply this study’s method to larger-scale fermentation in order to understand sweet corn’s application as a refreshment substrate in larger-capacity and longer operations.

Acknowledgement

        The authors acknowledge the Ministry of Education and Culture, Indonesia. This work was supported by the Islamic Development Bank Supporting Program, University of Jember, Indonesia. We have no conflicts of interest to declare.

Supplementary Material
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R1-CE-4354-20201125005109.jpg Figure 1
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