• 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
References

Al-Shorgani, N.K.N., Kalil, M.S., Yusoff, W.M.W., Hamid, A.A., 2018. Impact of pH and Butyric Acid on Butanol Production during Batch Fermentation using a New Local Isolate of Clostridium Acetobutylicum YM1. Saudi Journal of Biological Sciences, Volume 25(2), pp. 339–348

Ambarsari, H., Sonomoto, K., 2012. Enhanced Acetone, Butanol, and Ethanol Fermentation by Clostridium Accharoperbutylacetonicum N1-4 (ATCC 13564) in a Chemically Defined Medium: Effect of Iron and Initial pH on ABE Ratio. Microbiology Indonesia, Volume 6(4), pp. 139–147

Ambarsari, H., Sonomoto, K., 2015. Acetone-Butanol-Ethanol Fermentation for Bioenergy using Various Substrates in Defined TYA Media. Jurnal Energi Dan Lingkungan (Enerlink), Volume 11(1), pp. 49–60

Darmayanti, R.F., Amini, H.W., Rizkiana, M.F., Setiawan, F.A., Palupi, B., Rahmawati, I., Susanti, A., Fachri, B.A., 2019. Lignocellulosic Material from Main Indonesian Plantation Commodity as the Feedstock for Fermentable Sugar in Biofuel Production. ARPN Journal of Engineering and Applied Sciences, Volume 14(20), pp. 3524–3534

Darmayanti, R.F., Tashiro, Y., Noguchi, T., Gao, M., Sakai, K., Sonomoto, K., 2018. Novel Biobutanol Fermentation at a Large Extractant Volume Ratio using Immobilized Clostridium Saccharoperbutylacetonicum N1-4. Journal of Bioscience and Bioengineering, Volume 126(6), pp. 750–757

Devaux, A., Kromann, P., Ortiz, O., 2014. Potatoes for Sustainable Global Food Security. Potato Research, Volume 57(3), pp. 185–199

Febrianti, F., Syamsu, K., Rahayuningsih, M., 2017. Bioethanol Production from Tofu Waste by Simultaneous Saccharification and Fermentation (SSF) using Microbial Consortium. International Journal of Technology, Volume 8(5), pp. 898–908

Ferchichi, M., Crabbe, E., Hintz, W., Gil, G.-H., Almadidy, A., 2005. Influence of Culture Parameters on Biological Hydrogen Production by Clostridium saccharoperbutylacetonicum ATCC 27021. World Journal of Microbiology and Biotechnology, Volume 21(6), p. 855–862

Firgianti, G., Sunyoto, M., 2018. Karakterisasi Fisik dan Kimia Ubi Jalar Ungu (Ipomoea Batatas L ) Varietas Biang untuk Mendukung Penyediaan Bahan Baku Tepung Ubi Jalar Ungu (Physical and Chemical Characterization of Purple Sweet Potato (Ipomoea batatas) Biang Variety to Support the Feedstock Supply for Purple Sweet Potato Flour). In: Seminar Nasional Dies Natalis UNS Ke-42 (National Seminar of 42nd UNS Anniversary), Volume 2(1), pp. 104–110

Gao, M., Tashiro, Y., Wang, Q., Sakai, K., Sonomoto, K., 2016. High Acetone–Butanol–Ethanol Production in pH-Stat Co-Feeding of Acetate and Glucose. Journal of Bioscience and Bioengineering, Volume 122(2), pp. 176–182

Hastuti, N., Darmayanti, R.F., Hardiningtyas, S.D., Kanomata, K., Sonomoto, K., Goto, M., Kitaoka, T., 2019. Nanocellulose from Oil Palm Biomass to Enhance Microbial Fermentation of Butanol for Bioenergy Applications. BioResources, Volume 14(3), pp. 6936–6957

Jusuf, M., Ginting, E., 2014. The Prospects and Challenges of Sweet Potato as Bio-Ethanol Source in Indonesia. Energy Procedia, Volume 47, pp. 173–179

Kim, S.Y., Ryu, C.H., 1995. Studies on the Nutritional Components of Purple Sweet Potato(Ipomoea Batatas). Korean Journal of Food Science and Technology, Volume 27(5), pp. 819–825

Lapuerta, M., Ballesteros, R., Barba, J., 2017. Strategies to Introduce n-Butanol in Gasoline Blends. Sustainability (Switzerland), Volume 9(4), pp. 589–598

Li, H-g., Luo, W., Wang, Q., Yu, X-b., 2014. Direct Fermentation of Gelatinized Cassava Starch to Acetone , Butanol , and Ethanol using Clostridium Acetobutylicum Mutant Obtained by Atmospheric and Room Temperature Plasma. Applied Biochemistry and Biotechnology, Volume 172(7), pp. 3330–3341

Li, S.-Y., Srivastava, R., Suib, S.L., Li, Y., Parnas, R.S., 2011. Performance of Batch, Fed-Batch, and Continuous A–B–E Fermentation with pH-Control. Bioresource Technology, Volume 102(5), pp. 4241–4250

Li, S., Huang, L., Ke, C., Pang, Z., Liu, L., 2020. Pathway Dissection, Regulation, Engineering and Application: Lessons Learned from Biobutanol Production by Solventogenic Clostridia. Biotechnology for Biofuels, Volume 13(39), pp. 1–25

Li, T., Yan, Y., He, J., 2015. Enhanced Direct Fermentation of Cassava to Butanol by Clostridium Species Strain BOH3 in Cofactor-Mediated Medium. Biotechnology for Biofuels, Volume 8(166), pp. 1–12

Liu, K., Zheng, J., Wang, X., Chen, F., 2019. Effects of Household Cooking Processes on Mineral, Vitamin B, and Phytic Acid Contents and Mineral Bioaccessibility in Rice. Food Chemistry, Volume 280, pp. 59–64

McGill, C.R., Kurilich, A.C., Davignon, J., 2013. The Role of Potatoes and Potato Components in Cardiometabolic Health: A Review. Annals of Medicine, Volume 45(7), pp. 467–473

Merola, S.S., Tornatore, C., Marchitto, L., Valentino, G., Corcione, F.E., 2012. Experimental Investigations of Butanol-Gasoline Blends Effects on the Combustion Process in a SI Engine. International Journal of Energy and Environmental Engineering, Volume 3(6), pp. 1–14

Mukherjee, M., Sarkar, P., Goswami, G., Das, D., 2019. Regulation of Butanol Biosynthesis in Clostridium Acetobutylicum ATCC 824 under the Influence of Zinc Supplementation and Magnesium Starvation. Enzyme and Microbial Technology, Volume 129, 109352. https://doi.org/10.1016/j.enzmictec.2019.05.009

Muthayya, S., Sugimoto, J.D., Montgomery, S., Maberly, G.F., 2014. An Overview of Global Rice Production, Supply, Trade, and Consumption. Annals of the New York Academy of Sciences, Volume 1324(1), pp. 7–14

Oshiro, M., Hanada, K., Tashiro, Y., Sonomoto, K., 2010. Efficient Conversion of Lactic Acid to Butanol with pH-Stat Continuous Lactic Acid and Glucose Feeding Method by Clostridium Saccharoperbutylacetonicum. Applied Microbiology and Biotechnology, Volume 87(3), pp. 1177–1185

Qureshi, N., Singh, V., Liu, S., Ezeji, T.C., Saha, B.C., Cotta, M.A. 2014. Process Integration for Simultaneous Saccharification, Fermentation, and Recovery (SSFR): Production of Butanol from Corn Stover using Clostridium Beijerinckii P260. Bioresource Technology, Volume 154, pp. 222–228

Sari, F.K., Nurhayati., Djumarti., 2013. The Extraction of Starch Resistant from Three Local Varieties Potatoes as Prebiotic Candidates. Berkala Ilmiah Pertanian, Volume 1, pp. 38–42

Supramono, D., Jonathan., Haqqyana., Setiadi., Nasikin, M., 2016. Improving Bio-oil Quality Through Co-pyrolysis of Corn Cobs and Polypropylene in a Stirred Tank Reactor, International Journal of Technology, Volume 7(8), pp. 1382–1392

Surtinah., 2008. Waktu Panen yang Tepat Menentukan Kandungan Gula Biji Jagung Manis (Zea Mays Saccharata) (The Precise Harvesting Time to Determine Sugar Content of Sweet Corn Kernel (Zea mays saccharata)). Jurnal Ilmiah Pertanian, Volume 4(2), pp. 5–6

Szulczyk, K.R., 2010. Which Is a Better Transportation Fuel–Butanol or Ethanol?. International Journal of Energy and Environment, Volume 1(3), pp. 501–512

Tashiro, Y., Takeda, K., Kobayashi, G., Sonomoto, K., Ishizaki, A., Yoshino, S., 2004. High Butanol Production by Clostridium Saccharoperbutylacetonicum N1-4 in Fed-Batch Culture with pH-Stat Continuous Butyric Acid and Glucose Feeding Method. Journal of Bioscience and Bioengineering, Volume 98(4), 263–268

Tashiro, Y., Yoshida, T., Noguchi, T., Sonomoto, K., 2013. Recent Advances and Future Prospects for Increased Butanol Production by Acetone-Butanol-Ethanol Fermentation. Engineering in Life Sciences, Volume 13(5), pp. 432–445

Thang, V.H., Kanda, K., Kobayashi, G., 2010. Production of Acetone–Butanol–Ethanol (ABE) in Direct Fermentation of Cassava by Clostridium Saccharoperbutylacetonicum N1-4. Applied Biochemistry and Biotechnology, Volume 161(1), pp. 157–170

Xue, C., Zhao, J., Lu, C., Yang, S.T., Bai, F., Tang, I.C., 2012. High-Titer n-Butanol Production by Clostridium Acetobutylicum JB200 in Fed-Batch Fermentation with Intermittent Gas Stripping. Biotechnology and Bioengineering, Volume 109(11), pp. 2746–2756

Yuliansyah, A.T., Putri, C.O., Clarasinta, B.D., Nonaka, M., 2019. TGA Investigation of CO2 Gasification of Hydrothermally Treated Biomass (Corn Cob - Coconut Shell Mixture). International Journal of Technology, Volume 10(6), pp. 1166–1173

Zhao, T., Tashiro, Y., Zheng, J., Sakai, K., Sonomoto, K., 2018. Semi-Hydrolysis with Low Enzyme Loading Leads to Highly Effective Butanol Fermentation. Bioresource Technology, Volume 264, pp. 335–342

Zhao, T., Yasuda, K., Tashiro, Y., Darmayanti, R.F., Sakai, K., Sonomoto, K., 2019. Semi-Hydrolysate of Paper Pulp without Pretreatment Enables a Consolidated Fermentation System with in Situ Product Recovery for the Production of Butanol. Bioresource Technology, Volume 278, pp. 57–65