Mutational enhancement of Aspergillus niger Tiegh. for higher cellulase production comparable to Trichoderma species in solid-state fermentation

Harjeet Singh; Komal Janiyani; Ajit Gangawane

doi:10.7324/JABB.2025.200551

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Abstract

The present study aimed to obtain a fungal isolate and enhance its cellulolytic activity through various mutational techniques, focusing on achieving results comparable to those obtained from extensively researched isolates of Trichoderma sp. An Aspergillus niger Tiegh. strain was obtained from environmental samples collected in Vadodara, Gujarat, and its cellulolytic-enzyme production was assessed and found to be akin to Trichoderma viride and Trichoderma reesei. Utilizing rice straw in solid-state fermentation (SSF), this research evaluated optimal conditions for enzyme activity at diverse pH and temperatures for both wild-type and mutant strains modified employing NTG (Methylnitronitrosoguanidine), EMS (Ethyl Methane Sulphonate), and UV radiation treatments. NTG-induced mutations significantly enhanced enzyme yields, particularly at pH 5 and 27°C. The NTG-treated A. niger mutants exhibited remarkable increases in endoglucanase activity, achieving 30.36 Units/mL, which corresponds to an increase of 22.91 Units/mL (4.12-fold) compared to the untreated strain. Similarly, T. viride and T. reesei showed significant increases to 29.18 and 25.58 Units/mL, respectively. Also, A. niger NTG-treated mutants showed avicelase activity of 245.73 ± 14.9 Units/mLat pH 5 and 27°C, compared to untreated strains with 85.62 Units/mL, representing a 2.43-fold increase. Moreover, the addition of cellulose significantly boosted enzymatic activity, increasing endoglucanase activity in A. niger from 4.76 Units/mL to 6.29 Units/mL.The findings of this study highlight the capacity of NTG-mutagenesis to enhance the production of cellulase. This opens up encouraging prospects for optimizing the utilization of enzymes in industrial settings and promoting sustainable waste management via bioconversion technologies.

Keyword: Cellulase Mutagenesis Solid-state fermentation Sustainable waste management

Citation:

Singh H, Janiyani K, Gangawane A. Mutational enhancement of Aspergillus niger Tiegh. for higher cellulase production comparable to Trichoderma species in solid-state fermentation. J App Biol Biotech. 2025. Online First. http://doi.org/10.7324/JABB.2025.200551

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1. Kumar A, Agrawal A. Recent trends in solid waste management status, challenges, and potential for the future Indian cities - A review. Curr Res Environ Sustain. 2020;2. https://doi.org/10.1016/j.crsust.2020.100011

2. Badhan AK, Chadha BS, Kaur J, Saini HS, Bhat MK. Production of multiple xylanolytic and cellulolytic enzymes by thermophilic fungus Myceliophthora sp. IMI 387099. Bioresour Technol. 2007;98:504-10. https://doi.org/10.1016/j.biortech.2006.02.009

3. Sohail M, Siddiqi R, Ahmad A, Khan SA. Cellulase production from Aspergillus niger MS82: effect of temperature and pH. N Biotechnol. 2009;25:437-41. https://doi.org/10.1016/j.nbt.2009.02.002

4. Pérez J, Muñoz-Dorado J, De La Rubia T, Martínez J. Biodegradation and biological treatments of cellulose, hemicellulose and lignin: an overviewPérez. Int Microbiol. 2002;5:53-63. https://doi.org/10.1007/s10123-002-0062-3

5. Grover D, Chaudhry S. Ambient air quality changes after stubble burning in rice-wheat system in an agricultural state of India. Environ Sci Pollut Res. 2019;26:20550-9. https://doi.org/10.1007/s11356-019-05395-5

6. Gadde B, Bonnet S, Menke C, Garivait S. Air pollutant emissions from rice straw open field burning in India, Thailand and the Philippines. Environ Pollut. 2009;157:1554-8. https://doi.org/10.1016/j.envpol.2009.01.004

7. He Q, Zhao X, Lu J, Zhou G, Yang H, Gao W, et al. Impacts of biomass-burning on aerosol properties of a severe haze event over Shanghai. Particuology. 2015;20:52-60. https://doi.org/10.1016/j.partic.2014.11.004

8. Venkataraman C, Habib G, Kadamba D, Shrivastava M, Leon J.-F, Crouzille B, et al. Emissions from open biomass burning in India: Integrating the inventory approach with high-resolution Moderate Resolution Imaging Spectroradiometer (MODIS) active-fire and land cover data. Glob Biogeochem Cycles. 2006;20. https://doi.org/10.1029/2005GB002547

9. Deswal D, Khasa YP, Kuhad RC. Optimization of cellulase production by a brown rot fungus Fomitopsis sp. RCK2010 under solid state fermentation. Bioresour Technol. 2011;102:6065-72. https://doi.org/10.1016/j.biortech.2011.03.032

10. Gao J, Weng H, Zhu D, Yuan M, Guan F, Xi Y. Production and characterization of cellulolytic enzymes from the thermoacidophilic fungal Aspergillus terreus M11 under solid-state cultivation of corn stover. Bioresour Technol. 2008;99:7623-9. https://doi.org/10.1016/j.biortech.2008.02.005

11. Singhania RR, Sukumaran RK, Patel AK, Larroche C, Pandey A. Advancement and comparative profiles in the production technologies using solid-state and submerged fermentation for microbial cellulases. Enzyme Microb Technol. 2010;46:541-9. https://doi.org/10.1016/j.enzmictec.2010.03.010

12. Sari SLA, Setyaningsih R, Wibowo NFA. Isolation and screening of cellulolytic fungi from Salacca edulis leaf litter. Biodiversitas. 2017;18:1282-8. https://doi.org/10.13057/biodiv/d180355

13. Wen Z, Liao W, Chen S. Production of cellulase/β-glucosidase by the mixed fungi culture Trichoderma reesei and Aspergillus phoenicis on dairy manure. Process Biochem. 2005;40:3087-94. https://doi.org/10.1016/j.procbio.2005.03.044

14. Angela S, Oziniel R, Ignatious N, Thembekile N. Diversity of cellulase- and xylanase-producing filamentous fungi from termite mounds. J Yeast Fungal Res. 2019;10:15-29. https://doi.org/10.5897/JYFR2019.0189

15. Sakpetch P, H-Kittikun A, Chandumpai A. Isolation and screening of potential lignocellulolytic microorganisms from rubber bark and other agricultural residues. Walailak J Sci Technol. 2017;14:953-67.

16. Muthezhilan R, Ashok R, Jayalakshmi S. Production and optimization of thermostable alkaline xylanase by Penicillium oxalicum in solid state fermentation. J Microbiol. 2007:20-8.

17. Friedel JK, Munch JC, Fischer WR. Soil microbial properties and the assessment of available soil organic matter in a haplic luvisol after several years of different cultivation and crop rotation. Soil Biol Biochem. 1996;28:479-88. https://doi.org/10.1016/0038-0717(95)00188-3

18. Emmerling C, Udelhoven T, Schröder D. Response of soil microbial biomass and activity to agricultural de-intensification over a 10 year period. Soil Biol Biochem. 2001;33:2105-14. https://doi.org/10.1016/S0038-0717(01)00143-2

19. Sunitha VH, Devi DN, Srinivas C. Extracellular Enzymatic activity of endophytic fungal strains isolated from medicinal plants. World J Agric Sci. 2013;9:1-9.

20. Yan C, Yan SS, Jia TY, Dong SK, Ma CM, Gong ZP. Decomposition characteristics of rice straw returned to the soil in northeast China. Nutr Cycl Agroecosyst. 2019;114:211-24. https://doi.org/10.1007/s10705-019-09999-8

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