Potential of Secondary Metabolites of Species Associated with Terrestrial and Marine Origins.

species produce versatile secondary metabolites (SMs), including terpenoids, fatty acids, polyketides, steroids, and alkaloids. These structurally diverse SMs exhibit a wide range of biological activities, including cytotoxic, antifungal, antibacterial, antiviral, antioxidant, anti-inflammatory, and phytotoxic activities, which could be exploited in the medical, agricultural, and other modern industries. This review comprehensively covers the production and biological potencies of isolated natural products from the genus associated with terrestrial and marine origins.

Heterologous Expression, Purification and Characterization of an Alkalic Thermophilic β-Mannanase CcMan5C from .

A endo-1,4-β-mannanase (CcMan5C) gene was cloned from and heterologously expressed in , and the recombinant enzyme was purified by Ni-affinity chromatography and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). CcMan5C hydrolyzed only locust bean gum galactomannan (LBG) but not α-mannan from or Avicel cellulose, oat spelt xylan, or laminarin from . CcMan5C exhibited distinctive catalytic features that were different from previously reported β-mannanases.

Molecular Mechanism by Which the GATA Transcription Factor CcNsdD2 Regulates the Developmental Fate of under Dark or Light Conditions.

Coprinopsis cinerea has seven homologs of the Aspergillus nidulans transcription factor NsdD. Of these, CcNsdD1 and CcNsdD2 from show the best identities of 62 and 50% to A. nidulans NsdD, respectively.

The extracellular β-glucosidase BGL2 has two variants with different molecular sizes and hydrolytic activities in the stipe or pilei of .

Two variants of extracellular β-glucosidase (BGL2) were purified from the stipe and pilei of . In the stipe, BGL2 was a monomeric protein with an apparent molecular mass of approximately 220 kDa, representing a mature full-length peptide of BGL2. However, in the pilei, the apparent molecular mass of BGL2 was only approximately 120 kDa, consisting of the 60 kDa N-terminal fragment and 55 kDa C-terminal fragment.

Accumulation and cross-linkage of β-1,3/1,6-glucan lead to loss of basal stipe cell wall extensibility in mushroom Coprinopsis cinerea.

The mature basal stipe of mushroom Coprinopsis cinerea loses wall extensibility. We found that an endo-β-1,3-glucanase ENG from C. cinerea could restore mature basal stipe wall extensibility via pretreatment such that the ENG-pretreated basal stipe walls could be induced to extend by chitinase ChiIII.

A novel endo-β-1,6-glucanase from the mushroom Coprinopsis cinerea and its application in studying of cross-linking of β-1,6-glucan and the wall extensibility in stipe cell walls.

We previously reported that chitinases reconstituted heat-inactivated stipe cell wall extension in a steady and continuous extension profile by cleaving chitins cross-linked to various polysaccahrides, whereas, endo-β-1,3-glucanases reconstituted heat-inactivated stipe wall extension in a profile of an initially fast extension and subsequent termination of extension due to its degradation of β-1,3-glucan but not other polysaccharides such as β-1,6-glucans cross-linked to chitins. Thus, a novel endo-β-1,6-glucanase, GH30A, from Coprinopsis cinerea was cloned and characterized to study cross-linking of β-1,6-glucan and wall extensibility in stipe walls. GH30A had higher activity and better thermophilicity than reported β-1,6-glucanases.

Comparative study of β-glucan-degrading enzymes from Coprinopsis cinerea for their capacities to induce stipe cell wall extension.

We previously reported endo-β-1,3-glucanase ENG in combination with β-glucosidase BGL2 at low concentration induced stipe cell wall extension. This study further explored ENG could be replaced by endo-β-1,3(4)-glucanase ENG16A in combination with BGL2 to induce stipe cell wall extension; similarly, BGL2 could be replaced by β-glucosidase BGL1 to cooperate with ENG to induce stipe cell wall extension. However, ENG could not be replaced by exo-β-1,3-glucanase EXG in combination with BGL2 to induce stipe cell wall extension, although EXG alone released higher level of soluble sugars from the stipe cell walls during the reconstituted wall extension than that released from the stipe cell walls by a combination of ENG16A or ENG and BGL2 or BGL1, which was different from chitinase-mediated stipe cell wall extension.

Heterologous expression and characterization of a novel chitin deacetylase, CDA3, from the mushroom Coprinopsis cinerea.

The chitin deacetylase CDA3 from C. cinerea deacetylated chitin-oligosaccharides with dp ≥ 2. Since CDA3 firstly removed the intermediate acetyl group of (GlcNAc), it was an endo-acting deacetylase.

β-Glucosidase BGL1 from Exhibits a Distinctive Hydrolysis and Transglycosylation Activity for Application in the Production of 3-O-β-d-Gentiobiosyl-d-laminarioligosaccharides.

We previously reported that β-glucosidase BGL1 at low concentration (15 μg mL) from exhibited hydrolytic activity only toward laminarioligosaccharides but not toward cellooligosaccharides and gentiobiose. This study shows that BGL1 at high concentration (200 μg mL) also hydrolyzed cellobiose and gentiobiose, which accounted for only 0.83 and 2.

Glucanase-Induced Stipe Wall Extension Shows Distinct Differences from Chitinase-Induced Stipe Wall Extension of Coprinopsis cinerea.

This study reports that a high concentration of the endo-β-1,3-glucanase ENG (200 μg ml) induced heat-inactivated stipe wall extension of , whereas a high concentration of the extracellular β-glucosidase BGL2 (1,000 μg ml) did not; however, in combination, low concentrations of ENG (25 μg ml) and BGL2 (260 μg ml) induced heat-inactivated stipe cell wall extension. In contrast to the previously reported chitinase-reconstituted stipe wall extension, β-1,3-glucanase-reconstituted heat-inactivated stipe cell wall extension initially exhibited a fast extension rate that quickly decreased to zero after approximately 60 min; the stipe cell wall extension induced by a high concentration of β-1,3-glucanase did not result in stipe breakage during measurement, and the inner surfaces of glucanase-reconstituted extended cell walls still remained as amorphous matrices that did not appear to have been damaged. These distinctive features of the β-1,3-glucanase-reconstituted wall extension may be because chitin chains are cross-linked not only to the nonreducing termini of the side chains and the backbones of β-1,6 branched β-1,3-glucans but also to other polysaccharides.

Chitinases Play a Key Role in Stipe Cell Wall Extension in the Mushroom .

The elongation growth of the mushroom stipe is a characteristic but not well-understood morphogenetic event of basidiomycetes. We found that extending native stipe cell walls of were associated with the release of -acetylglucosamine and chitinbiose and with chitinase activity. Two chitinases among all detected chitinases from , ChiE1 and ChiIII, reconstituted heat-inactivated stipe wall extension and released -acetylglucosamine and chitinbiose.

Characteristics, transcriptional patterns and possible physiological significance of glycoside hydrolase family 16 members in Coprinopsis cinerea.

The glycoside hydrolase (GH) 16 family of Coprinopsis cinerea includes 15 members distributed in four subgroups (A1, A2, B and D) by phylogenetic analysis. The expression patterns match well with the requirement of wall-softening in the germination of basidiospores, hyphal growth and branching, primordium formation, stipe elongation, pileus expansion and autolysis. Remarkably, expression levels of different GH16 members varied with different morphogenetic events.

HPAEC-PAD and Q-TOF-MS/MS analysis reveal a novel mode of action of endo-β-1,3(4)-d-glucanase Eng16A from coprinopsis cinerea on barley β-glucan.

We previously reported that an endo-β-1,3(4)-d-glucanase, Eng16A, from C. cinerea shows a higher degradation activity toward barley β-glucan than laminarin. HPAEC-PAD and Q-TOF-MS/MS analyses show that Eng16A-digestion products of barley β-glucan not only contain some oligosaccharides with (1 → 3)-β-linkage adjacent to the reducing end, which is consistent with β-1,3(4)-glucanase-digestion products, but also include some oligosaccharides containing (1 → 4)-β-linkage adjacent to the reducing end which is consistent with cellulase-digestion products.

A novel thermophilic exochitinase ChiEn3 from Coprinopsis cinerea exhibits a hyperhydrolytic activity toward 85% deacetylated chitosan and a significant application to preparation of chitooligosaccharides from the chitosan.

ChiEn3 from Coprinopsis cinerea was characterized as an exo-acting chitinase with a processivity. ChiEn3 hydrolyzed only soluble chitin and exhibited a hyperhydrolytic activity toward 85% deacetylated chitosan which was 33.6-fold higher than its hydrolytic activity toward glycol chitin.

ChiE1 from Coprinopsis cinerea is Characterized as a Processive Exochitinase and Revealed to Have a Significant Synergistic Action with Endochitinase ChiIII on Chitin Degradation.

Fruiting bodies that exhibit strong autolysis of Coprinopsis cinerea are a good resource for the chitinolytic system. In this study, a new Chitinase ChiE1 from C. cinerea was cloned, heterologously expressed, and characterized.

Gene cloning, heterologous expression and characterization of a Coprinopsis cinerea endo-β-1,3(4)-glucanase.

A gene coding endo-β-1,3(4)-glucanase (ENG16A) was cloned from Coprinopsis cinerea and heterologously expressed in Pichia pastoris. ENG16A only acts on substrates containing β-1,3 glycosidic bonds but not on substrates containing only β-1,4- or β-1,6-glycosidic bonds. Interestingly, compared to the activity of this enzyme towards carboxymethyl (CM)-pachyman containing only β-1,3-glycosidic bonds, its activity towards barley β-glucan containing both β-1,3-glycosidic and β-1,4-glycosidic bonds was increased by 64.

Purification, characterization and physiological significance of a chitinase from the pilei of Coprinopsis cinerea fruiting bodies.

We purified a chitinase from pilei extractions of Coprinopsis cinerea fruiting bodies by ammonium sulfate precipitation and CM Sepharose cation exchange chromatography. MALDI-TOF/TOF MS analysis characterized this purified chitinase as a putative class V chitinase, ChiB1. ChiB1 hydrolyzed colloidal chitin and chitosan, whereas it did not hydrolyze chitin powder.

Purification, characterization and function analysis of an extracellular β-glucosidase from elongating stipe cell walls in Coprinopsis cinerea.

A β-glycoside hydrolase was isolated from cell walls material in Coprinopsis cinerea elongating stipes. By analysis of SDS-PAGE, MALDI-TOF/TOF MS and substrate specificity, this enzyme was characterized as an extracellular β-glucosidase which is a trimer consisting of three homosubunits. β-Glucosidase did not degrade β-glucans with modified ends, whereas it hydrolyzed various β-glucans with free ends and related oligosaccharides with β-1,3-, β-1,4- or β-1,6-linkages.

Silica-supported sulfonic acid-functionalized ionic liquid coated with [bmim][PF6] as a scavenger for the synthesis of amides.

The methodology of using a silica gel-supported functionalized ionic liquid as a scavenger in the purification of parallel synthesis products was demonstrated. Silica-supported sulfonic acid-functional ionic liquid was synthesized by etherification, aminate, and quaternary aminate from activated silica gel and 3-chloropropyl trimethoxysilane, imidazole, and 1,4-butanesultone, which was followed by acidification using trifluoromethanesulfonic acid and anion exchange with potassium hexafluorophosphate. A conventional ionic liquid, 1-butyl-3-methylimidazolium hexafluorophosphate was then used to coat the surface of the silica gel.