Abstract
Lignocellulosic biomasses are extensively used by researchers to produce a variety of renewable bioproducts. This research described an environment-friendly technique of xylitol production by an adapted strain of Candida tropicalis from areca nut hemicellulosic hydrolysate, produced through enzymatic hydrolysis. To enhance the activity of xylanase enzymes, lime and acid pretreatment was conducted to make biomass more amenable for saccharification. To improve the efficiency of enzymatic hydrolysis, saccharification parameters like xylanase enzyme loading were varied. Results exposed that the highest yield (g/g) of reducing sugar, about 90%, 83%, and 15%, were achieved for acid-treated husk (ATH), lime-treated husk (LTH), and raw husk (RH) at an enzyme loading of 15.0 IU/g. Hydrolysis was conducted at a substrate loading of 2% (w/V) at 30 °C, 100 rpm agitation, for 12 h hydrolysis time at pH 4.5 to 5.0. Subsequently, fermentation of xylose-rich hemicellulose hydrolysate was conducted with pentose utilizing the yeast Candida tropicalis to produce xylitol. The optimum concentration of xylitol was obtained at about 2.47 g/L, 3.83 g/L, and 5.88 g/L, with yields of approximately 71.02%, 76.78%, and 79.68% for raw fermentative hydrolysate (RFH), acid-treated fermentative hydrolysate (ATFH), and lime-treated fermentative gydrolysate (LTFH), respectively. Purification and crystallization were also conducted to separate xylitol crystals, followed by characterization like X-ray diffraction (XRD) and scanning electron microscopy (SEM) analysis. Results obtained from crystallization were auspicious, and about 85% pure xylitol crystal was obtained.
Graphical Abstract
Similar content being viewed by others
Data Availability
The data that support the findings of this study are available from the corresponding authors, upon reasonable request.
Abbreviations
- ATEH:
-
Acid-treated enzymatic hydrolysate
- ATFH:
-
Acid-treated fermentative hydrolysate
- ATH:
-
Acid-treated husk
- CMC:
-
Carboxy methyl cellulose
- CrI:
-
Crystallinity index
- DNS:
-
Di-nitrosalicylic acid
- FESEM:
-
Field emission scanning electron microscopy
- FXC:
-
Fermented xylitol crystal
- HMF:
-
Hydroxymethylfurfural
- LTEH:
-
Lime-treated enzymatic hydrolysate
- LTFH:
-
Lime-treated fermentative hydrolysate
- LTH:
-
Lime-treated husk
- MB:
-
Methylene blue
- MBS:
-
Methylene blue staining
- MTCC:
-
Microbial Type Culture Collection
- OPEFB:
-
Oil palm empty fruit bunch
- REH:
-
Raw enzymatic hydrolysate
- RFH:
-
Raw fermentative hydrolysate
- RH:
-
Raw husk
- SSFH:
-
Synthetic solution fermentative hydrolysate
- XCCX:
-
Xylitol crystal from commercial xylitol
- XRD:
-
X-ray diffraction
- YPD :
-
Yeast extract, peptone, and dextrose
References
Ranjithkumar, M., Ravikumar, R., Sankar, M. K., Kumar, M. N., & Thanabal, V. (2017). An effective conversion of cotton waste biomass to ethanol: A critical review on pretreatment processes. Waste and Biomass Valorization, 8(1), 57–68. https://doi.org/10.1007/S12649-016-9563-8/TABLES/2
Keshav, P. K., Naseeruddin, S., & Rao, L. V. (2016). Improved enzymatic saccharification of steam exploded cotton stalk using alkaline extraction and fermentation of cellulosic sugars into ethanol. Bioresource Technology, 214, 363–370. https://doi.org/10.1016/J.BIORTECH.2016.04.108
Da Silva, A. S. A., Inoue, H., Endo, T., Yano, S., & Bon, E. P. S. (2010). Milling pretreatment of sugarcane bagasse and straw for enzymatic hydrolysis and ethanol fermentation. Bioresource Technology, 101(19), 7402–7409. https://doi.org/10.1016/J.BIORTECH.2010.05.008
de Araújo, C. K. C., de Oliveira Campos, A., de Araújo Padilha, C. E., de Sousa Júnior, F. C., do Nascimento, R. J. A., de Macedo, G. R., & dos Santos, E. S. (2017). Enhancing enzymatic hydrolysis of coconut husk through Pseudomonas aeruginosa AP 029/GLVIIA rhamnolipid preparation. Bioresource Technology, 237, 20–26. https://doi.org/10.1016/J.BIORTECH.2017.03.178
Vardhan, H., Mahato, R. B., Sasmal, S., & Mohanty, K. (2022). Production of xylose from pre-treated husk of areca nut. Journal of Natural Fibers, 19(1), 131–144. https://doi.org/10.1080/15440478.2020.1731905
Goli, J. K., & Hameeda, B. (2021). Production of xylitol and ethanol from acid and enzymatic hydrolysates of Typha latifolia by Candida tropicalis JFH5 and Saccharomyces cerevisiae VS3. Biomass Conversion and Biorefinery, 1–11. https://doi.org/10.1007/S13399-021-01868-1/FIGURES/6
Felipe Hernández-Pérez, A., de Arruda, P. V., Sene, L., da Silva, S. S., Kumar Chandel, A., de AFelipi Almeida, M., & Das, G. (2019). Xylitol bioproduction: State-of-the-art, industrial paradigm shift, and opportunities for integrated biorefineries. Critical Reviews in Biotechnology, 39(7), 924–943. https://doi.org/10.1080/07388551.2019.1640658
Unrean, P., & Ketsub, N. (2018). Integrated lignocellulosic bioprocess for co-production of ethanol and xylitol from sugarcane bagasse. Industrial Crops and Products, 123, 238–246. https://doi.org/10.1016/J.INDCROP.2018.06.071
Sluiter, A., Hames, B., Ruiz, R., Scarlata, C., Sluiter, J., Templeton, D., & Crocker, D. (2008). Determination of structural carbohydrates and lignin in biomass laboratory analytical procedure (LAP) issue date: 7/17/2005. Retrieved from www.nrel.gov
Meddeb-Mouelhi, F., Moisan, J. K., & Beauregard, M. (2014). A comparison of plate assay methods for detecting extracellular cellulase and xylanase activity. Enzyme and Microbial Technology, 66, 16–19. https://doi.org/10.1016/J.ENZMICTEC.2014.07.004
Martins, M., Henriques, M., Azeredo, J., Rocha, S. M., Coimbra, M. A., & Oliveira, R. (2007). Morphogenesis control in Candida albicans and Candida dubliniensis through signaling molecules produced by planktonic and biofilm cells. Eukaryotic Cell, 6(12), 2429–2436. https://doi.org/10.1128/EC.00252-07/ASSET/372A4F99-9812-40E2-9A8A-D1DB245418CE/ASSETS/GRAPHIC/ZEK0120730350003.JPEG
Martínez, E. A., Canettieri, E. V. C., Bispo, J. A., Giulietti, M., de Almeida Silva, J. B., Converti, A., & de Almeida, J. B. (2015). Strategies for xylitol purification and crystallization: A review. Separation Science and Technology, 50, 2087–2098. https://doi.org/10.1080/01496395.2015.1009115
Goli, J. K., & Hameeda, B. (2021). Production of xylitol and ethanol from acid and enzymatic hydrolysates of Typha latifolia by Candida tropicalis JFH5 and Saccharomyces cerevisiae VS3. Biomass Conversion and Biorefinery, 1, 1–11. https://doi.org/10.1007/S13399-021-01868-1/FIGURES/6
Martín, C., De Moraes Rocha, G. J., Dos Santos, J. R. A., De Albuquerque Wanderley, M. C., & Gouveia, E. R. (2012). Enzyme loading dependence of cellulose hydrolysis of sugarcane bagasse. Química Nova, 35(10), 1927–1930. https://doi.org/10.1590/S0100-40422012001000007
Vardhan, H., Sasamal, S., & Mohanty, K. (2022). Fermentation process optimisation based on ANN and RSM for xylitol production from areca nut husk followed by xylitol crystal characterisation. Process Biochemistry, 122(P2), 146–159. https://doi.org/10.1016/j.procbio.2022.10.005
Sun, X. F., Xu, F., Sun, R. C., Fowler, P., & Baird, M. S. (2005). Characteristics of degraded cellulose obtained from steam-exploded wheat straw. Carbohydrate Research, 340(1), 97–106. https://doi.org/10.1016/J.CARRES.2004.10.022
Julie Chandra, C. S., George, N., & Narayanankutty, S. K. (2016). Isolation and characterization of cellulose nanofibrils from arecanut husk fibre. Carbohydrate Polymers, 142, 158–166. https://doi.org/10.1016/J.CARBPOL.2016.01.015
Li, Z., Guo, X., Feng, X., & Li, C. (2015). An environment friendly and efficient process for xylitol bioconversion from enzymatic corncob hydrolysate by adapted Candida tropicalis. Chemical Engineering Journal, 263, 249–256. https://doi.org/10.1016/J.CEJ.2014.11.013
Chosdu, R., Hilmy, N., ErizalErlinda, T. B., & Abbas, B. (1993). Radiation and chemical pretreatment of cellulosic waste. Radiation Physics and Chemistry, 42(4–6), 695–698. https://doi.org/10.1016/0969-806X(93)90354-W
Bak, J. S., Ko, J. K., Han, Y. H., Lee, B. C., Choi, I. G., & Kim, K. H. (2009). Improved enzymatic hydrolysis yield of rice straw using electron beam irradiation pretreatment. Bioresource Technology, 100(3), 1285–1290. https://doi.org/10.1016/J.BIORTECH.2008.09.010
Zhao, X., Zhang, L., & Liu, D. (2012). Biomass recalcitrance. Part I: The chemical compositions and physical structures affecting the enzymatic hydrolysis of lignocellulose. Biofuels Bioproducts and Biorefining, 6(4), 465–482. https://doi.org/10.1002/BBB.1331
Cai, C., Bao, Y., Li, F., Pang, Y., Lou, H., Qian, Y., & Qiu, X. (2020). Using highly recyclable sodium caseinate to enhance lignocellulosic hydrolysis and cellulase recovery. Bioresource Technology, 304, 122974. https://doi.org/10.1016/J.BIORTECH.2020.122974
Rafał Łukajtis, Piotr Rybarczyk *, Karolina Kucharska, D. K.-Ł., & Edyta Słupek, K. W. and M. K. ´nski. (2018). Optimization of saccharification conditions of lignocellulosic biomass under alkaline. https://doi.org/10.3390/en11040886
Tavares, J. M., Duarte, L. C., Amaral-Collaço, M. T., & Gírio, F. M. (2000). The influence of hexoses addition on the fermentation of d-xylose in Debaryomyces hansenii under continuous cultivation. Enzyme and Microbial Technology, 26(9–10), 743–747. https://doi.org/10.1016/S0141-0229(00)00166-6
Rivas, B., Domínguez, J. M., Domínguez, H., & Parajó, J. C. (2002). Bioconversion of posthydrolysed autohydrolysis liquors: An alternative for xylitol production from corn cobs. Enzyme and Microbial Technology, 31(4), 431–438. https://doi.org/10.1016/S0141-0229(02)00098-4
Tada, K., Horiuchi, J. I., Kanno, T., & Kobayashi, M. (2004). Microbial xylitol production from corn cobs using Candida magnoliae. Journal of bioscience and bioengineering, 98(3), 228–230. https://doi.org/10.1016/S1389-1723(04)00273-7
de Albuquerque, T. L., Gomes, S. D. L., Marques, J. E., da Silva, I. J., & Rocha, M. V. P. (2015). Xylitol production from cashew apple bagasse by Kluyveromyces marxianus CCA510. Catalysis Today, 255, 33–40. https://doi.org/10.1016/J.CATTOD.2014.10.054
Li, M., Meng, X., Diao, E., & Du, F. (2012). Xylitol production by Candida tropicalis from corn cob hemicellulose hydrolysate in a two-stage fed-batch fermentation process. Journal of Chemical Technology & Biotechnology, 87(3), 387–392. https://doi.org/10.1002/JCTB.2732
Carvalheiro, F., Duarte, L. C., Medeiros, R., & Gírio, F. M. (2007). Xylitol production by Debaryomyces hansenii in brewery spent grain dilute-acid hydrolysate: Effect of supplementation. Biotechnology letters, 29(12), 1887–1891. https://doi.org/10.1007/S10529-007-9468-5
Baek, S. C., & Kwon, Y. J. (2007). Optimization of the pretreatment of rice straw hemicellulosic hydrolyzates for microbial production of xylitol. Biotechnology and Bioprocess Engineering, 12(4), 404–409. https://doi.org/10.1007/BF02931063
Mardawati, E., Maharani, N., Wira, D. W., Harahap, B. M., Yuliana, T., & Sukarminah, E. (2020). Xylitol production from oil palm empty fruit bunches (OPEFB) via simultaneous enzymatic hydrolysis and fermentation process. Journal of Industrial and Information Technology in Agriculture, 2(1), 29–36. https://doi.org/10.24198/JIITA.V2I1.25064
Harahap, B. M., & Kresnowati, M. T. A. P. (2018). Moderate pretreatment of oil palm empty fruit bunches for optimal production of xylitol via enzymatic hydrolysis and fermentation. Biomass Conversion and Biorefinery, 8(2), 255–263. https://doi.org/10.1007/S13399-017-0299-X/TABLES/3
Wang, L., Yang, M., Fan, X., Zhu, X., Xu, T., & Yuan, Q. (2011). An environmentally friendly and efficient method for xylitol bioconversion with high-temperature-steaming corncob hydrolysate by adapted Candida tropicalis. Process Biochemistry, 46(8), 1619–1626. https://doi.org/10.1016/J.PROCBIO.2011.05.004
Cao, N. J., Tang, R., Gong, C. S., & Chen, L. F. (1994). The effect of cell density on the production of xylitol fromd-xylose by yeast. Applied Biochemistry and Biotechnology, 45(1), 515–519. https://doi.org/10.1007/BF02941826
Vandeska, E., Amartey, S., Kuzmanova, S., & Jeffries, T. (1995). Effects of environmental conditions on production of xylitol by Candida boidinii. World Journal of Microbiology and Biotechnology, 11(2), 213–218. https://doi.org/10.1007/BF00704652
Parajó, J. C., Domínguez, H., & Domínguez, J. M. (1998). Biotechnological production of xylitol. Part 1: Interest of xylitol and fundamentals of its biosynthesis. Bioresource Technology, 65(3), 191–201. https://doi.org/10.1016/S0960-8524(98)00038-8
Martínez, E. A., de Almeida e Silva, J. B., Giulietti, M., & Solenzal, A. I. N. (2007). Downstream process for xylitol produced from fermented hydrolysate. Enzyme and Microbial Technology, 40(5), 1193–1198. https://doi.org/10.1016/J.ENZMICTEC.2006.09.003
Marques Júnior, J. E., & Rocha, M. V. P. (2021). Development of a purification process via crystallization of xylitol produced for bioprocess using a hemicellulosic hydrolysate from the cashew apple bagasse as feedstock. Bioprocess and Biosystems Engineering, 44(4), 713–725. https://doi.org/10.1007/S00449-020-02480-9/FIGURES/9
Kresnowati, M. T. A. P., Regina, D., Bella, C., Wardani, A. K., & Wenten, I. G. (2019). Combined ultrafiltration and electrodeionization techniques for microbial xylitol purification. Food and Bioproducts Processing, 114, 245–252. https://doi.org/10.1016/J.FBP.2019.01.005
Wei, J., Yuan, Q., Wang, T., & Wang, L. (2010). Purification and crystallization of xylitol from fermentation broth of corncob hydrolysates. Frontiers of Chemical Engineering in China, 4(1), 57–64. https://doi.org/10.1007/S11705-009-0295-1
Sampaio, F. C., Passos, F. M. L., Passos, F. J. V., De Faveri, D., Perego, P., & Converti, A. (2006). Xylitol crystallization from culture media fermented by yeasts. Chemical Engineering and Processing: Process Intensification, 45(12), 1041–1046. https://doi.org/10.1016/J.CEP.2006.03.012
Deng, L. H., Tang, Y., & Liu, Y. (2014). Detoxification of corncob acid hydrolysate with SAA pretreatment and xylitol production by immobilized Candida tropicalis. Scientific World Journal. https://doi.org/10.1155/2014/214632
Misra, S., Raghuwanshi, S., & Saxena, R. K. (2013). Evaluation of corncob hemicellulosic hydrolysate for xylitol production by adapted strain of Candida tropicalis. Carbohydrate Polymers, 92(2), 1596–1601. https://doi.org/10.1016/J.CARBPOL.2012.11.033
Funding
Present research work vides grant no. (BT/PR16747/NER/95/271/2015) was financially supported by the Department of Biotechnology (DBT), Government of India.
Author information
Authors and Affiliations
Contributions
All authors contributed to the study’s conception and design. Material preparation, data collection, and analysis were performed by Harsha Vardhan, Soumya Sasmal, and Kaustubha Mohanty. The first draft of the manuscript was written by Harsha Vardhan, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Ethics Approval
Not applicable.
Conflict of Interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Vardhan, H., Sasamal, S. & Mohanty, K. Xylitol Production by Candida tropicalis from Areca Nut Husk Enzymatic Hydrolysate and Crystallization. Appl Biochem Biotechnol 195, 7298–7321 (2023). https://doi.org/10.1007/s12010-023-04469-y
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-023-04469-y