Optimization of Thermotolerant Cellulase from Streptomyces sp. M127-1 with Response Surface Methodology using Solid-State Fermentation

Year 2024, , 217 - 227, 27.02.2024

Muzaffer Binek Elif Esin Hameş

https://doi.org/10.35414/akufemubid.1336051

Abstract

The study aims to optimise cellulase that can be used in composting from actinomycete isolate using the solid-state fermentation method with Response Surface Methodology and its partial characterisation. Five isolates (M127-1, M127-2B, 1M1, M45-1 and M6c-1) were previously determined to be candidate cellulase producers, were investigated for cellulase production by solid-state fermentation, and isolate M127-1 was selected for further studies. For solid substrate selection, solid-state fermentation was performed by isolate M127-1 in media-prepared combinations of different solid substrates (sawdust, soy flour, malt grass and wheat bran), and the highest enzyme activity was found to be wheat bran-malt grass (BM) medium (0.704 U/ml). Then, moisture, inoculum size, and incubation time were optimised with the central composite design for cellulase optimisation using BM medium. Optimum cellulase production conditions were determined as 79.93% moisture, 8.58 days incubation time and 8.38 (v/w) inoculum size, and it was observed that the enzyme activity increased by 1.8 times. The optimum pH of the enzyme was 6, and its temperature was 60C. The isolate was identified as Streptomyces sp. M127-1 by 16SrDNA sequence analysis. In conclusion, the thermotolerant cellulase production at high temperatures was optimised from Streptomyces sp. M127-1 through solid-state fermentation using cost-effective substrates.

Keywords

Cellulose, Streptomyces, Composting, Solid state fermentation, Response surface methodology

Project Number

FYL-2020-22356

Katı Hal Fermantasyonu Kullanılarak Streptomyces sp. M127-1’den Elde Edilen Termotolerant Selülazın Yanıt Yüzeyi Metodolojisi ile Optimizasyonu

Abstract

Çalışma, kompostlamada kullanılabilecek selülazın aktinomiset izolatından katı hal fermantasyon yöntemiyle Yanıt Yüzeyi Metodolojisi (RSM) kullanılarak optimizasyonunu ve enzimin kısmi karakterizasyonunu amaçlamaktadır. Selülaz üreticisi olduğu belirlenmiş 5 izolat (M127-1, M127-2B, 1M1, M45-1 ve M6c-1) katı hal fermantasyonu ile selülaz üretimi açısından değerlendirilmiş ve izolat M127-1 ileri çalışmalar için seçilmiştir. Katı substrat seçimi için farklı substratların (talaş, malt çimi, soya unu ve buğday kepeği) kombinasyonları ile hazırlanan ortamlarda izolat M127-1 ile katı hal fermantasyonu gerçekleştirilmiş ve en iyi enzim aktivitesinin buğday kepeği-malt çimi (BM) (0,704 U/ml) ortamında olduğu belirlenmiştir. Ardından BM ortamı kullanılarak selülaz üretim optimizasyonu için nem, aşı miktarı ve inkübasyon süresi merkezi kompozit tasarımı kullanılarak optimize edilmiştir. Optimum selülaz üretim koşulları % 79,93 nem, 8,58 gün inkübasyon süresi ve 8,38 (v/w) aşı miktarı olarak belirlenmiş ve enzim aktivitesinin 1,8 kat arttığı görülmüştür. Enzimin optimum pH’sı 6 ve optimum sıcaklığı 60C olarak belirlenmiştir. 16SrDNA dizi analizi ile izolat, Streptomyces sp. M127-1 olarak tanımlanmıştır. Sonuç olarak yüksek sıcaklıkta aktif selülazın uygun maliyetli substratlar kullanılarak katı hal fermantasyonu ile Streptomyces sp. M127-1’den istatistiksel üretim optimizasyonu gerçekleştirilmiştir.

Keywords

Selülaz, Streptomyces, Kompostlama, Katı hal fermentasyonu, Yanıt yüzey metodolojisi

Supporting Institution

Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

FYL-2020-22356

Thanks

Bu çalışma Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü FYL-2020-22356 nolu proje ile desteklenmiştir. Yazarlar çalışmaya olan desteklerinden dolayı Prof. Dr. Sait SARGIN’a teşekkür ederler.

References

• Abdul Manan, M., Webb, C., 2017. Modern microbial solid state fermentation technology for future biorefineries for the production of added-value products. Biofuel Research Journal, 16, 730-740. https://doi.org/10.18331/BRJ2017.4.4.5
• Antony, J., 2023. Design of experiment for engineers and scientist. Elsevier. Ayilara, M.S., Olanrewaju, O.S., Babalola, O.O., Odeyemi, O., 2020. Waste management through composting: Challenges and potentials. Sustainability, 12(11), 4456. https://doi.org/10.3390/su12114456
• Bezerra, M.A., Santelli, R.E., Oliveira, E.P., Villar, L.S., Escaleira, L.A. 2008. Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta, 76(5), 965-977. https://doi.org/10.1016/j.talanta.2008.05.019
• Bhatti, A.A., Haq, S., Bhat, R.A., 2017. Actinomycetes benefaction role in soil and plant health. Microbial pathogenesis, 111, 458-467. https://doi.org/10.1016/j.micpath.2017.09.036
• Bostubayeva, M., Baimbetova, E., Makenova, M., Shumenova, N., Sarmanova, R., Nauanova, A., 2023. Screening and evaluation of potential microbial bio-activators used in sewage sludge composting. Caspian Journal of Environmental Sciences, 1-9. https://doi.org/10.22124/CJES.2023.6936
• Box, G.E., Hunter, W.H., Hunter, S. 1978. Statistics for experimenters (Vol. 664). New York: John Wiley and Sons. Celaya-Herrera, S., Casados-Vázquez, L.E., Valdez-Vazquez, I., Barona-Gómez, F., Bideshi, D.K., Barboza-Corona, J.E., 2021. A cellulolytic Streptomyces sp. isolated from a highly oligotrophic niche shows potential for hydrolyzing agricultural wastes. BioEnergy Research, 14, 333-343. https://doi.org/10.1007/s12155-020-10174-z
• Chen, X., Cheng, W., Li, S., Tang, X., Wei, Z., 2021. The “quality” and “quantity” of microbial species drive the degradation of cellulose during composting. Bioresource Technology, 320, 124425. https://doi.org/10.1016/j.biortech.2020.124425
• Crawford, D. L., 1978. Lignocellulose decomposition by selected Streptomyces strains. Applied and Environmental Microbiology, 35(6), 1041-1045. https://doi.org/10.1128/aem.35.6.1041-1045.1978
• Crawford, D.L., Lynch, J.M., Whipps, J.M., Ousley, M. A., 1993. Isolation and characterization of actinomycete antagonists of a fungal root pathogen. Applied and Environmental Microbiology, 59(11), 3899-3905. https://doi.org/10.1128/aem.59.11.3899-3905.1993
• Cuesta, G., García-de-la-Fuente, R., Abad, M., Fornes, F., 2012. Isolation and identification of actinomycetes from a compost-amended soil with potential as biocontrol agents. Journal of Environmental Management, 95, S280-S284. https://doi.org/10.1016/j.jenvman.2010.11.023
• Danso, B., Ali, S.S., Xie, R., Sun, J., 2022. Valorisation of wheat straw and bioethanol production by a novel xylanase-and cellulase-producing Streptomyces strain isolated from the wood-feeding termite, Microcerotermes species. Fuel, 310, 122333. https://doi.org/10.1016/j.fuel.2021.122333
• Demir, T., Hameş, E.E., Öncel, S.S., Vardar-Sukan, F., 2015. An optimization approach to scale up keratinase production by Streptomyces sp. 2M21 by utilizing chicken feather. International Biodeterioration & Biodegradation, 103, 134-140. https://doi.org/10.1016/j.ibiod.2015.04.025
• Duangupama, T., Pittayakhajonwut, P., Intaraudom, C., Suriyachadkun, C., Sirirote, P., He, Y.W., Thawai, C., 2022. Streptomyces sennicomposti sp. nov., an actinomycete isolated from compost of Senna siamea (Lam.). International Journal of Systematic and Evolutionary Microbiology, 72(4), 005320. https://doi.org/10.1099/ijsem.0.005320
• Ghose, T. K., 1987. Measurement of cellulase activities. Pure and Applied Chemistry, 59(2), 257-268. https://doi.org/10.1351/pac198759020257
• Hobbs, G., Frazer, C.M., Gardner, D.C., Cullum, J.A., Oliver, S.G., 1989. Dispersed growth of Streptomyces in liquid culture. Applied Microbiology and Biotechnology, 31, 272-277. https://doi.org/10.1007/BF00258408
• Insam, H., and De Bertoldi, M., 2007. Microbiology of the composting process. In Waste Management Series Vol. 8,. Elsevier, pp. 25-48. https://doi.org/10.1016/S1478-7482(07)80006-6
• Kausar, H., Sariah, M., Mohd Saud, H., Zahangir Alam, M., Razi Ismail, M., 2011. Isolation and screening of potential actinobacteria for rapid composting of rice straw. Biodegradation, 22, 367-375. https://doi.org/10.1007/s10532-010-9407-3
• Khan, M.N., Luna, I.Z., Islam, M.M., Sharmeen, S., Salem, K.S., Rashid, T.U., Zaman, A., Haque, P., Rahman, M.M., 2016. Cellulase in waste management applications. In New and Future Developments in Microbial Biotechnology and Bioengineering, Elsevier, pp. 237-256. https://doi.org/10.1016/B978-0-444-63507-5.00021-6
• Kocak, F.O., Tanir, S.G.E., Cetin, A.K., Degirmenci, L., 2023. Simulatenous evaluation of composting experiments and metagenome analyses to illuminate the effect of Streptomyces spp. on organic matter degradation. World Journal of Microbiology and Biotechnology, 39(3), 70. https://doi.org/10.1007/s11274-023-03516-4
• Korsa, G., Konwarh, R., Masi, C., Ayele, A., Haile, S., 2023. Microbial cellulase production and its potential application for textile industries. Annals of Microbiology, 73(1), 13. https://doi.org/10.1186/s13213-023-01715-w
• Kumar, M., Kumar, P., Das, P., Solanki, R., Kapur, M.K., 2022. Proactive role of Streptomyces spp. in plant growth stimulation and management of chemical pesticides and fertilizers. International Journal of Environmental Science and Technology, 19(10), 10457-10476. https://doi.org/10.1007/s13762-021-03473-1
• Korn-Wendisch F, Kutzner HJ (1992) The family Streptomycetaceae. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes. Springer, New York, pp 921–995.
• Lee, Y.J., Kim, B.K., Lee, B.H., Jo, K.I., Lee, N.K., Chung, C.H., Lee, Y-C., Lee, J.W., 2008. Purification and characterization of cellulase produced by Bacillus amyoliquefaciens DL-3 utilizing rice hull. Bioresource Technology, 99(2), 378-386. https://doi.org/10.1016/j.biortech.2006.12.013
• Manan, M.A., and Webb, C., 2017. Design aspects of solid state fermentation as applied to microbial bioprocessing. Journal of Applied Biotechnology and Bioengineering 4(1), 91. https://doi.org/10.15406/jabb.2017.04.00094
• Mandenius, C.F., and Brundin, A., 2008. Bioprocess optimization using design‐of‐experiments methodology. Biotechnology Progress, 24(6), 1191-1203. https://doi.org/10.1002/btpr.67
• Mazumder, P., Akhil, P.M., Khwairakpam, M., Mishra, U., Kalamdhad, A.S., 2021. Enhancement of soil physico-chemical properties post compost application: Optimization using Response Surface Methodology comprehending Central Composite Design. Journal of Environmental Management, 289, 112461. https://doi.org/10.1016/j.jenvman.2021.112461
• Mendoza-Cal, A., Cuevas-Glory, L., Lizama-Uc, G., & Ortiz-Vázquez, E., 2010. Naringinase production from filamentous fungi using grapefruit rind in solid state fermentation. African Journal of Microbiology Research, 4(19), 1964-1969.
• Palaniveloo, K., Amran, M.A., Norhashim, N.A., Mohamad-Fauzi, N., Peng-Hui, F., Hui-Wen, L., Kai-Lin, Y., Jiale, L., Chian-Yee, M. G., Jing-Yi, L., Gunasekaran, B.,Razak, S.A., 2020. Food waste composting and microbial community structure profiling. Processes, 8(6), 723. https://doi.org/10.3390/pr8060723
• Patel, A.K., Singhania, R.R., Sim, S.J., & Pandey, A., 2019. Thermostable cellulases: current status and perspectives. Bioresource Technology, 279, 385-392. https://doi.org/10.1016/j.biortech.2019.01.049
• Raimbault, M. 1998. General and Microbiological Aspects of Solid Substrate Fermentation Electronic Journal of Biotechnology, 1(3), 26-27. http://dx.doi.org/10.4067/S0717-34581998000300007
• Ramachandra, M., Crawford, D.L., Hertel, G., 1988. Characterization of an extracellular lignin peroxidase of the lignocellulolytic actinomycete Streptomyces viridosporus. Applied and Environmental Microbiology, 54(12), 3057-3063. https://doi.org/10.1128/aem.54.12.3057-3063.1988
• Sánchez, Ó. J., Ospina, D. A., & Montoya, S. (2017). Compost supplementation with nutrients and microorganisms in composting process. Waste management, 69, 136-153. https://doi.org/10.1016/j.wasman.2017.08.012
• Samuel, M.S., Govarthan, M., Selvarajan, E., 2022. A comprehensive review on strategic study of cellulase producing marine actinobacteria for biofuel applications. Environmental Research, 214, 114018. https://doi.org/10.1016/j.envres.2022.114018
• Sargın, S, ve Göksungur, Y., 2007. Çeşitli tarımsal atık ve yan ürünlerin katı kültür fermantasyonu ile laktik asit üretiminde kullanılabilirliklerinin incelenmesi. Ege Üniversitesi Ziraat Fakültesi Dergisi, 44(3), 89-99.
• Sargın, S., ve Öngen, G., 2003. Kanatlı yemi katkısı olarak kullanılan ksilanaz enziminin katı kültür fermantasyon yöntemi ile üretiminde ölçek büyütme çalışmaları. Ege Üniversitesi Ziraat Fakültesi Dergisi, 40(3).
• Sun, C., Wei, Y., Kou, J., Han, Z., Shi, Q., Liu, L., Sun, Z., 2021. Improve spent mushroom substrate decomposition, bacterial community and mature compost quality by adding cellulase during composting. Journal of Cleaner Production, 299, 126928. https://doi.org/10.1016/j.jclepro.2021.126928
• Tamura, K., Stecher, G., Kumar, S., 2021. MEGA11: molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution, 38(7), 3022-3027. https://doi.org/10.1093/molbev/msab120
• Topatan, Z.Ş., and Katı H., 2022. Screening of actinomycetes from Cystoseira barbata (Stackhouse) C. Agardh compost for their enzyme and antibacterial activities. Trakya University Journal of Natural Sciences. 23(2): 113-124. https://doi.org/10.23902/trkjnat.1059974
• Tuomela, M., Vikman, M., Hatakka, A., Itävaara, M., 2000. Biodegradation of lignin in a compost environment: a review. Bioresource Technology, 72(2), 169-183. https://doi.org/10.1016/S0960-8524(99)00104-2
• Zainudin, M.H.M., Zulkarnain, A., Azmi, A.S., Muniandy, S., Sakai, K., Shirai, Y., Hassan, M.A., 2022. Enhancement of agro-industrial waste composting process via the microbial inoculation: a brief review. Agronomy, 12(1), 198. https://doi.org/10.3390/agronomy12010198