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Developing and evaluating lignocellulolytic hyper producing deregulated strains of Mycothermus thermophilus for hydrolysis of lignocellulosics

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Abstract

An indigenous thermophilic fungal strain of Mycothermus thermophilus (Syn. Scytalidium thermophilum) designated as CM 4 T was subjected to a combination of cyclic mutagenesis and intra-specific protoplast fusion to induce heterokaryosis and diploidization for developing deregulated cellulase hyper-producer strain. The developed strain (23 U) produced 14.60, 45.50, 82.80, 3.28, and 264 U/ml of CBH, β-glucosidase, CMCase, Fpase, and xylanase at fermenter level, which were 8.6, 2.54, 6.58, 2.37, and 6.58 folds improved, respectively, in comparison to wild-type strain CM 4 T. Furthermore, the developed mutants produced cellulases constitutively during fed-batch (flask mode) using glucose feed at 0.375 g carbon/l/h. The creA gene and CreA protein structures of mutant 23 U showed truncated zinc finger motif linking it to the catabolite repression-resistant phenotype. The hydrolytic potential of cellulase produced of developed strain 23 U was evaluated using acid- and alkali-treated rice straw and bagasse at 10 % substrate loading rate. Furthermore, studies showed that hydrolytic efficiency of the cellulases produced by mutant 23 U can be further enhanced by custom designing of the cocktails that comprised of lignocellulolytic enzymes of mutant 23 U and different thermophilic/thermotolerant fungal strains.

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Acknowledgement

The financial support from the Department of Biotechnology, India, for carrying out research project “Developing fungal strains for enzyme production through mutagenesis, protoplast fusion and over expression using A. niger citrate synthase promoter” (BT/PR15271/PBD/26/509/2015) and from the Department of Science and Technology, India, for carrying out research project “Manipulation of transcriptional factors for developing hypercellulase producing strains for bioconversion of lignocellulosic biomass” (SR/WOS-A/LS-260/2018) is duly acknowledged.

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  1. Neha Basotra and Baljit Kaur contributed equally to this work.

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  1. Department of Microbiology, Guru Nanak Dev University, Amritsar, Punjab, 143005, India

    Neha Basotra, Baljit Kaur, Yashika Raheja, Dhruv Agrawal, Gaurav Sharma & Bhupinder Singh Chadha

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  1. Neha Basotra
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Basotra, N., Kaur, B., Raheja, Y. et al. Developing and evaluating lignocellulolytic hyper producing deregulated strains of Mycothermus thermophilus for hydrolysis of lignocellulosics. Biomass Conv. Bioref. 13, 5059–5071 (2023). https://doi.org/10.1007/s13399-021-01539-1

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