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

Abstract Background Major 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. Results Hydrolysis 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. Conclusions The 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.

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Penicillium Janthinellum NCIM-1366 Shows Improved Biomass Hydrolysis Performance Over Trichoderma Reesei RUT-C30, and Secretome Analysis Reveals a Larger Number of CAZymes with Higher Induction Levels.

10.21203/rs.3.rs-53399/v1 ◽

2020 ◽

Author(s):

AthiraRaj Sreeja-R ◽

Meera Christopher ◽

Prajeesh Kooloth-Valappil ◽

Rajasree Kuni-Parambil ◽

Digambar Vittal Gokhale ◽

...

Keyword(s):

Trichoderma Reesei ◽

Feedback Inhibition ◽

Total Concentration ◽

Relative Proportion ◽

Biomass Hydrolysis ◽

Extracellular Medium ◽

Penicillium Janthinellum ◽

Enzyme Preparations ◽

Industrial Grade ◽

Rut C30

Abstract BackgroundMajor cost of bioethanol is attributed to enzymes employed in biomass hydrolysis. Lignocellulolytic 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 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 10-fold 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 more number of CAZymes and a higher relative abundance of cellulases upon cellulose induction in the fungus.ConclusionsThe results revealed the ability 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 could be the possible reasons for its performance advantage in biomass hydrolysis.

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

10.21203/rs.3.rs-53399/v2 ◽

2020 ◽

Author(s):

AthiraRaj Sreeja-R ◽

Meera Christopher ◽

Prajeesh Kooloth-Valappil ◽

Rajasree Kuni-Parambil ◽

Digambar Vittal Gokhale ◽

...

Keyword(s):

Trichoderma Reesei ◽

Feedback Inhibition ◽

Total Concentration ◽

Relative Proportion ◽

Biomass Hydrolysis ◽

Extracellular Medium ◽

Penicillium Janthinellum ◽

Enzyme Preparations ◽

Industrial Grade ◽

Rut C30

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 10-fold 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.

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Whole Genome Sequence and CAZyme distribution of the cellulase hyper producing filamentous fungus Penicillium janthinellum NCIM 1366

10.1101/2021.06.17.448855 ◽

2021 ◽

Author(s):

Meera K Christopher ◽

AthiraRaj Sreeja-Raju ◽

Prajeesh K Kooloth-Valappil ◽

Amith Abraham ◽

Digambar Vitthal Gokhale ◽

...

Keyword(s):

De Novo ◽

Cellulase Production ◽

Whole Genome Sequence ◽

Lignocellulose Degradation ◽

Carbohydrate Active Enzymes ◽

Biomass Hydrolysis ◽

Penicillium Janthinellum ◽

Trichoderma Reesei Rut C30 ◽

Hydrolysis Of ◽

Rut C30

Penicillium janthinellum NCIM 1366, capable of secreting cellulases that are highly efficient in the hydrolysis of lignocellulosic biomass, was sequenced to understand its cellulolytic machinery. De novo sequencing and assembly revealed a 37.6 Mb genome encoding 11,848 putative proteins, 93% of which had significant BLAST-P hits. The majority of the top hits (those with over 60% UniProt identity) belonged to P. brasilianum. Carbohydrate active enzymes (CAZymes) and other enzymes involved in lignocellulose degradation were also predicted from this strain and compared with those of the industrial workhorse of cellulase production- Trichoderma reesei RUT-C30. The comparison showed that the fungus encodes a far higher number of CAZYmes (422) as compared to T. reesei RUT-C30 (244), which gives a plausible explanation for its overall effectiveness in biomass hydrolysis. An analysis of the secreted CAZymes and annotated ligninases identified 216 predicted proteins which may be directly involved in the breakdown of lignocellulose

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On-site integrated production of cellulases and β-glucosidases by Trichoderma reesei Rut C30 using steam-pretreated sugarcane bagasse

10.1101/461012 ◽

2018 ◽

Cited By ~ 3

Author(s):

Marcella Fernandes de Souza ◽

Elba Pinto da Silva Bon ◽

Ayla Sant’ Ana da Silvab

Keyword(s):

Carbon Source ◽

Lignocellulosic Biomass ◽

Trichoderma Reesei ◽

Sugarcane Bagasse ◽

Enzyme Preparation ◽

Integrated Approach ◽

Aspergillus Awamori ◽

Pretreated Sugarcane Bagasse ◽

Hydrolysis Of ◽

Rut C30

AbstractThe high cost of commercial cellulases still hampers the economic competitiveness of the production of fuels and chemicals from lignocellulosic biomasses. This cost may be decreased by the on-site production of cellulases with the integrated use of the lignocellulosic biomass as carbon source. This integrated approach was evaluated in the present study whereby steam-pretreated sugarcane bagasse (SPSB) was used as carbon source for the production of cellulases by Trichoderma reesei Rut C30 and the produced enzymes were subsequently used for SPSB hydrolysis. An enzyme preparation with a high cellulase activity, of 1.93 FPU/mL, was obtained, and a significant β-glucosidase activity was achieved in buffered media, indicating the importance of pH control during enzyme production. The hydrolysis of SPSB with the laboratory-made mixture resulted in a glucose yield of 80%, which was equivalent to those observed for control experiments using commercial enzymes. Even though the supplementation of this mixture with external β-glucosidase from Aspergillus awamori was found to increase the initial hydrolysis rates, it had no impact on the final hydrolysis yield. It was shown that SPSB is a promising carbon source for the production of cellulases and β-glucosidases by T. reesei Rut C30 and that the enzyme preparation obtained is effective for the hydrolysis of SPSB, supporting the on-site integrated approach to decrease the cost of the enzymatic hydrolysis of lignocellulosic biomass.

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