Penicillium janthinellum NCIM1366 shows improved biomass hydrolysis and a larger number of CAZymes with higher induction levels over Trichoderma reesei RUT-C30

Athiraraj Sreeja-Raju,Meera Christopher,Meena Sankar,Prajeesh Kooloth-Valappil,Ashok Pandey,Rajeev K Sukumaran,Rajasree Kuni-Parambil,Amith Abraham,Digambar Vittal Gokhale

doi:10.1186/s13068-020-01830-9

Abstract

BackgroundMajor cost of bioethanol is attributed to enzymes employed in biomass hydrolysis. Biomass hydrolyzing enzymes are predominantly produced from the hyper-cellulolytic mutant filamentous fungus Trichoderma reesei RUT-C30. Several decades of research have failed to provide an industrial grade organism other than T. reesei, capable of producing higher titers of an effective synergistic biomass hydrolyzing enzyme cocktail. Penicillium janthinellum NCIM1366 was reported as a cellulase hyper producer and a potential alternative to T. reesei, but a comparison of their hydrolytic performance was seldom attempted.ResultsHydrolysis of acid or alkali-pretreated rice straw using cellulase enzyme preparations from P. janthinellum and T. reesei indicated 37 and 43% higher glucose release, respectively, with P. janthinellum enzymes. A comparison of these fungi with respect to their secreted enzymes indicated that the crude enzyme preparation from P. janthinellum showed 28% higher overall cellulase activity. It also had an exceptional tenfold higher beta-glucosidase activity compared to that of T. reesei, leading to a lower cellobiose accumulation and thus alleviating the feedback inhibition. P. janthinellum secreted more number of proteins to the extracellular medium whose total concentration was 1.8-fold higher than T. reesei. Secretome analyses of the two fungi revealed higher number of CAZymes and a higher relative abundance of cellulases upon cellulose induction in the fungus.ConclusionsThe results revealed the ability of P. janthinellum for efficient biomass degradation through hyper cellulase production, and it outperformed the established industrial cellulase producer T. reesei in the hydrolysis experiments. A higher level of induction, larger number of secreted CAZymes and a high relative proportion of BGL to cellulases indicate the possible reasons for its performance advantage in biomass hydrolysis.

Highlights

Major cost of bioethanol is attributed to enzymes employed in biomass hydrolysis
Cellulases are mainly divided into three major groups based on their mode of action on cellulose: (1) endoglucanases, which randomly cleave internal β-1, 4 linkages in cellulose chain generating free ends; (2) cellobiohydrolases, which act in a processive manner on either reducing or non-reducing ends of the cellulose chain, releasing cellobiose as the major product and (3) beta-glucosidases that hydrolyze cellobiose into glucose [5]
P. janthinellum secretes higher amount of proteins compared to T. reesei Cellulose is a large polymer and utilization of it requires secretion of enzyme by microorganisms to process it outside the cell, so that the simple sugars derived from its breakdown can be taken inside

Summary

Introduction

Major cost of bioethanol is attributed to enzymes employed in biomass hydrolysis. Biomass hydrolyz‐ ing enzymes are predominantly produced from the hyper-cellulolytic mutant filamentous fungus Trichoderma reesei RUT-C30. Degradation of lignocelluloses is generally carried out by Sreeja‐Raju et al Biotechnol Biofuels (2020) 13:196 a set of enzymes including ligninases, hemicellulases, cellulases and other accessory enzymes. Their relative proportions and quantities can determine the efficiency of hydrolysis [2]. The major sources of lignocellulose-degrading enzymes are filamentous fungi, mainly the genera Trichoderma, Aspergillus and Penicillium [4]. With cellulose being the major component, cellulases play an important role in the degradation process. Cellulases are mainly divided into three major groups based on their mode of action on cellulose: (1) endoglucanases, which randomly cleave internal β-1, 4 linkages in cellulose chain generating free ends; (2) cellobiohydrolases, which act in a processive manner on either reducing or non-reducing ends of the cellulose chain, releasing cellobiose as the major product and (3) beta-glucosidases that hydrolyze cellobiose into glucose [5]

Objectives

Methods

Results

Discussion

Conclusion