Penicillium : The next emerging champion for cellulase production

Vaishnav, Neha; Singh, Anusuiya; Adsul, Mukund; Dixit, Pooja; Sandhu, Simranjeet Kaur; Mathur, Anshu S.; Puri, S.K.; Singhania, Reeta Rani

doi:10.1016/j.biteb.2018.04.003

Cited by 98 publications

(44 citation statements)

References 96 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Cellulase mixtures of Penicillium spp. are known to be rich in β-glucosidase [2], they are found in GH1 and GH3 families in P. echinulatum 2HH. Those proteins belonging to GH1 are probably intracellular enzymes, whereas five of nine GH3 β-glucosidases contain a signal peptide and are probably secreted into the medium.…”

Section: P Echinulatum Cazyomementioning

confidence: 99%

“…Some Penicillium species have been highlighted due to its superiority over existing enzyme producers, especially due to the production of balanced cellulolytic cocktails rich in β-glucosidases. Consequently, these enzymatic mixtures result in improved enzymatic hydrolysis yields [2]. P. echinulatum is widely studied for biofuels production from lignocellulosic biomass, mainly agricultural residues, such as sugar-cane bagasse and elephant grass [3,4,5,6].…”

mentioning

confidence: 99%

See 1 more Smart Citation

CAZyome and Sugar Transportome Unveil Singular Aspects of the Lignocellulolytic Enzyme System of Penicillium echinulatum 2HH

Lenz

Balbinot

Oliveira

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: The production of bioethanol using lignocellulosic feedstocks has proved to be a cutting-edge technology. However, this technology faces limitations such as cost and yield of enzymatic systems, required to degrade efficiently the polysaccharides of plant cell walls. Penicillium echinulatum 2HH is a widely studied ascomycete, known by its efficient cellulolytic cocktails. One strategy to improve the saccharification yields for commercial exploitation is the design of hypersecreting strains. However, molecular knowledge about the lignocellulolytic system of this specific fungus is scarce. Understanding both lignocellulolytic and sugar uptake systems is essential to obtain industrial strains with adequate efficiency for bioethanol production. Results: We report a comprehensive in silico characterization of CAZymes and sugar transporters of P. echinulatum 2HH. The CAZyome reveals an outstanding repertoire of enzymes involved in plant biomass degradation. Among them, we highlight the cellulolytic enzyme system whose genes are predominantly orthologous to P. oxalicum 114-2, demonstrating the high similarity of these phylogenetically related enzyme producers. We also report a LPMO-type enzyme of the AA16 family described for the first time in these fungi. In addition to the well-known high activity of ß-glucosidases, we found that coding genes for the AA16, GH5-4 and GH45 families comprehend the main differences in the cellulolytic complexes of P. echinulatum 2HH and P. oxalicum 114-2, when both are compared to commercial producers. Our phylogenetic analysis of the sugar transportome suggests that P. echinulatum 2HH diversity and specificity of STs include eight major families with specificity to different groups of sugars. Finally, our phylogenetic analyses enabled the identification of several iBGLs and STs potentially involved in the accumulation of intracellular cellodextrins.Conclusions: Overall, both CAZyome and sugar transportome of P. echinulatum revealed new insights into the mechanisms underlying a flexible and highly functional metabolism to degrade plant biomass. Peculiarities found in our study help to highlight the cellulolytic complex of P. echinulatum 2HH, contributing to the commercial ascendance of Penicillium spp. as cellulolytic enzyme producers. Furthermore, the first phylogenetic classification of STs and iBGLs shed new light into the role of these genes regarding the preferred carbon source during fungal growth. Along these lines, these iBGLs and STs comprise valuable gene targets to understand the regulatory mechanisms underlying cellulolytic enzymes and to design hypersecreting strains with adequate efficiency for bioethanol production in large-scale.

show abstract

Section: P Echinulatum Cazyomementioning

confidence: 99%

mentioning

confidence: 99%

CAZyome and Sugar Transportome Unveil Singular Aspects of the Lignocellulolytic Enzyme System of Penicillium echinulatum 2HH

Lenz

Balbinot

Oliveira

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…This consequently brings about an attack on the pyranose ring of the glucose moieties at the C-1 or C-4 position, thereby destabilizing the adjacent glycosidic bond and breaking it by an elimination reaction (15, 16). These LPMOs targets the crystalline regions of the cellulose surface which are typically more recalcitrant to cellulase action and introduce nicks on the substrate surface thereby providing extra chain ends for glycoside hydrolases (GHs) to act (17). This action subsequently improves the reaction kinetics as well as enhance enzymatic synergy during saccharification (17, 18).…”

Section: Introductionmentioning

confidence: 99%

“…These LPMOs targets the crystalline regions of the cellulose surface which are typically more recalcitrant to cellulase action and introduce nicks on the substrate surface thereby providing extra chain ends for glycoside hydrolases (GHs) to act (17). This action subsequently improves the reaction kinetics as well as enhance enzymatic synergy during saccharification (17, 18). However, the critical drawback of not having a single strain which could secrete a balanced ratio of all the cellulase component (both hydrolytic and accessory) subsequently affects the overall enzyme cost required for biomass saccharification especially at high substrate concentration.…”

Section: Introductionmentioning

confidence: 99%

Insights into structure of Penicillium funiculosum LPMO and its synergistic saccharification performance with CBH1 on high substrate loading upon simultaneous overexpression

Ogunyewo

Randhawa

Gupta

et al. 2020

Preprint

View full text Add to dashboard Cite

1 synergistic saccharification performance with CBH1 on high substrate 2 loading upon simultaneous overexpression 3 4 5 Abstract 20 Lytic polysaccharide monooxygenases (LPMOs) are crucial industrial enzymes required in 21 the biorefinery industry as well as in natural carbon cycle. These enzymes known to possess 22 auxiliary activity are produced by numerous bacterial and fungal species to assist in the 23 degradation of cellulosic biomass. In this study, we annotated and performed structural 24 analysis of an uncharacterized thermostable LPMO from Penicillium funiculosum 25 (PfLPMO9) in an attempt to understand nature of this enzyme in biomass degradation. 26 PfLPMO9 exhibited 75% and 36% structural identity to Thermoascus aurantiacus 27 (TaLPMO9A) and Lentinus similis (LsLPMO9A), respectively. Analysis of the molecular 28 interactions during substrate binding revealed that PfLPMO9 demonstrated a higher binding 29 affinity with a ∆G free energy of -46 k kcal/mol when compared with that of TaLPMO9A (-30 31 kcal/mol). The enzyme was further found to be highly thermostable at elevated 31 temperature with a half-life of ~88 h at 50 o C. Furthermore, multiple fungal genetic 32 manipulation tools were employed to simultaneously overexpress this LPMO and 33 Cellobiohydrolase I (CBH1) in catabolite derepressed strain of Penicillium funiculosum, 34PfMig1 88 , in order to improve its saccharification performance towards acid pretreated wheat 35 straw (PWS) at 20% substrate loading. The resulting transformants showed ~200% and ~66% 36 increase in LPMO and Avicelase activities, respectively. While the secretomes of 37 individually overexpressed LPMO and CBH1-strains increased saccharification of PWS by 38 6% and 13%, respectively, over PfMig1 88 at same enzyme concentration, the simultaneous 39 overexpression of these two genes led to 20% increase in saccharification efficiency over 40 PfMig1 88 , which accounted for 82% saccharification of PWS at 20% substrate loading. 41 42 43 44 3 Importance 45Enzymatic hydrolysis of cellulosic biomass by cellulases continues to be a significant 46 bottleneck in the development of second-generation bio-based industries. While efforts are 47 being intensified at how best to obtain indigenous cellulase for biomass hydrolysis, the high 48 production cost of this enzyme remains a crucial challenge confronting its wide availability 49 for efficient utilization of cellulosic materials. This is because it is challenging to get an 50 enzymatic cocktail with balanced activity from a single host. This report provides for the first 51 time the annotation and structural analysis of an uncharacterized thermostable lytic 52 polysaccharide monooxygenase (LPMO) gene in Penicillium funiculosum and its impact in 53 biomass deconstruction upon overexpression in catabolite derepressed strain of P. 54 funiculosum. Cellobiohydrolase I (CBH1) which is the most important enzyme produced by 55 many cellulolytic fungi for saccharification of crystalline cellulose was further overexpressed 56 simultaneous...

show abstract

“…Although, desired enzymatic attributes vastly differ for different applications, high catalytic activity, stability at extreme environmental conditions and tolerance to end-product inhibition and high concentrations of salts and ions are generally among the most favorable characteristics of industrial cellulases [14][15][16].…”

mentioning

confidence: 99%

A novel high performance metagenome-derived alkali-thermostable endo-β-1,4-glucanase for lignocellulosic biomass hydrolysis in the harsh conditions

Ariaeenejad¹,

Sheykhabdolahzadeh²,

Maleki³

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Lignocellulosic biomass, is a great resource for the production of bio-energy and biobased material since it is largely abundant, inexpensive and renewable. The requirement of new energy sources has led to a wide search for novel effective enzymes to improve the exploitation of lignocellulose, among which the importance of thermostable and halotolerant cellulase enzymes with high pH performance is significant.Results: The primary aim of this study was to discover a novel alkali-thermostable endo-β-1,4glucanase from the sheep rumen metagenome. Using a multi-step in-silico analysis, primary candidates with desired properties were found and subjected to cloning, expression, and purification followed by functional and structural characterization. The enzymes' kinetic parameters, including V max , Km, and specific activity, were calculated. The PersiCel4 demonstrated its optimum activity at pH 8.5 and a temperature of 85°C and was able to retain more than 70% of its activity after 150 hours of storage at 85°C. Furthermore, this enzyme was able to maintain its catalytic activity in the presence of different concentrations of NaCl, MgCl 2 , CaCl 2 , and MnCl 2 . Our results showed that treatment with MnCl 2 could enhance the enzyme's activity by 89%. PersiCel4 was ultimately used for enzymatic hydrolysis of autoclave pretreated rice straw, the most abundant agricultural waste with rich cellulose content. In autoclave treated rice straw, enzymatic hydrolysis with the PersiCel4 increased the release of reducing sugar up to 260% after 72 hours in the harsh condition ( T= 85°C, pH = 8.5).Conclusion: Considering the urgent demand for stable cellulases that are operational on extreme temperature and pH conditions and due to several proposed distinctive characteristics of PersiCel4, it can be used in the harsh condition for bioconversion of lignocellulosic biomass. BackgroundLignocellulose is the principal constituent of the biomass and is the most abundant bio-renewable organic resource on earth and therefore is of significant interest to governments, researchers, and industries. Lignocellulosic biomass is composed of three biopolymers, hemicellulose, cellulose and lignin which, depending on the type of the biomass, are intermeshed and organized into complex

show abstract

Penicillium : The next emerging champion for cellulase production

Cited by 98 publications

References 96 publications

CAZyome and Sugar Transportome Unveil Singular Aspects of the Lignocellulolytic Enzyme System of Penicillium echinulatum 2HH

CAZyome and Sugar Transportome Unveil Singular Aspects of the Lignocellulolytic Enzyme System of Penicillium echinulatum 2HH

Insights into structure of Penicillium funiculosum LPMO and its synergistic saccharification performance with CBH1 on high substrate loading upon simultaneous overexpression

A novel high performance metagenome-derived alkali-thermostable endo-β-1,4-glucanase for lignocellulosic biomass hydrolysis in the harsh conditions

Contact Info

Product

Resources

About