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Production of Trichoderma harzianum K179 bioagent for maize diseases control: complete laboratory stage bioprocess development

Mitrović I, Vučurović D, Al-Ani LKT, Mitrović B, Bajić B, Dodić S, et al.

J Appl Microbiol, 2023 Jun 01;134(6).
PMID: 37279914 DOI: 10.1093/jambio/lxad115

AIMS: In order to be competitive on the market, the production of biopreparations needs to be optimized, modelled, and assessed in the early stages of its development. The aim of this paper was to optimize medium for the production of Trichoderma harzianum K179 biocontrol agent, to analyze its kinetics at enlarged laboratory scale and finally economic analysis of the production of this high-value product through simulation modelling.
METHODS AND RESULTS: The results showed that the bioprocess of T. harzianum K179 bioagent production in a laboratory bioreactor on the medium with optimal composition (dextrose 10 g l-1, soy flour 6.87 g l-1, K2HPO4 1.51 g l-1, KCl 0.5 g l-1, and MgSO4 × 7H2O 0.5 g l-1), at stirring speed of 1.75 × g and aeration intensity of 1.5 vvm, can be shortened from 96 to 36 h. The results of bioprocess economic analysis showed that with a 25-year project lifetime and an investment payback time of 7.58 years, this project represents an economically viable system.
CONCLUSIONS: Complete analysis of the bioprocess of T. harzianum K179 biocontrol agent production showed that the biologically produced preparation can be competitive on the market with synthetic preparations.

Matched MeSH terms: Hypocreales*
Pathogenicity of an Indigenous Strain of the Entomopathogenic Fungus Metarhizium anisopliae (Hypocreales: Clavicipitaceae) (MET-GRA4 Strain) as a Potential Biological Control Agent Against the Red Palm Weevil (Coleoptera: Dryophthoridae)

Ishak I, Ng LC, Haris-Hussain M, Jalinas J, Idris AB, Azlina Z, et al.

J Econ Entomol, 2020 02 08;113(1):43-49.
PMID: 31586213 DOI: 10.1093/jee/toz233

Metarhizium anisopliae Metchnikoff (Hypocreales: Clavicipitaceae) is a fungal pathogen that causes disease in various insect pests, and it can be exploited and developed as a biological control agent to combat the red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Dryophthoridae). The study on indigenous isolates is crucial especially for development of bioinsecticides in the future. The M. anisopliae strain called MET-GRA4 was tested for pathogenicity against adult red palm weevil and treated in vitro with different spore viabilities. The isolates exhibited pathogenicity with 100% mortality 21 d postinfection. The median lethal time (LT50) for 85% viable spores was 8.6 d, while 39% viable spores had an LT50 value of 21.37 d, with 92 and 16.6% mycosis, respectively. The species MET-GRA4 strain was molecularly characterized using ITS1 and ITS4 from pure culture (Isolate A), mass-produced spores (Isolate B), and infected red palm weevil cadavers (Isolate C). The DNA sequences obtained matched M. anisopliae sequences, with 99% similarity. This new isolate of M. anisopliae has potential as a targeted bioinsecticide for management of red palm weevil.

Matched MeSH terms: Hypocreales*
Fulltext Isolation of an obscure fungus, Parengyodontium album, from the blood of a severely neutropenic patient: The first reported case from Malaysia

Mohamed N, Ding CH, Wahab AA, Tzar MN, Hassan M

J Infect Dev Ctries, 2022 Oct 31;16(10):1668-1670.
PMID: 36332225 DOI: 10.3855/jidc.17016

Parengyodontium album is a very rarely encountered opportunistic fungal pathogen. A severely neutropenic 11-year-old boy with acute T-cell lymphoblastic leukemia/lymphoma was febrile and lethargic during his admission for elective chemotherapy. No cutaneous lesion or obvious source of infection was noted, and clinical examination was otherwise unremarkable. A blood specimen was sent for culture and fungal elements were visualized. Amphotericin B was administered empirically while awaiting fungal identification. Morphologically, a hyaline mould with thin septate hyphae plus smooth-walled conidiophores and conidiogenous cells arranged in whorls of up to four was cultured. Internal transcribed spacer region sequencing identified the fungus conclusively as P. album. Repeat blood culture was also positive for the same fungus. Following a two-week course of amphotericin B, fungemia clearance was attained.

Matched MeSH terms: Hypocreales*
Fulltext Inoculation and colonization of the entomopathogenic fungi, Isaria javanica and Purpureocillium lilacinum, in tomato plants, and their effect on seedling growth, mortality and adult emergence of Bemisia tabaci (Gennadius)

Sani I, Jamian S, Saad N, Abdullah S, Mohd Hata E, Jalinas J, et al.

PLoS One, 2023;18(5):e0285666.
PMID: 37216342 DOI: 10.1371/journal.pone.0285666

Entomopathogenic fungi (EPF) are natural enemies which affect insect population and have long been recognized as biological control agents against many insect pests. Some isolates have also been established as endophytes, benefiting their host plants without causing any symptoms or negative effects. Here we demonstrated two entomopathogenic fungal species, Isariajavanica (Frieder. & Bally) Samson & Hywel-jone 2005 and Purpureocillium lilacinum (Thom) Luangsa-ard, Hou-braken, Hywel-Jones & Samson (2011) as endophytes in tomato plants by using the seed inoculation method and examined their effect on plant growth, B. tabaci mortality, and adult emergence. Our study indicated that tomato seeds treated with a fungal suspension of I. javanica and P. lilacinum enabled their recovery from plant tissues (root, stem and leaf) up to 60 days after inoculation (DAI). Both endophytic isolates also caused significant mortality of adult B. tabaci on seedlings inoculated with, I. javanica (51.92±4.78%), and P. lilacinum (45.32±0.20%) compared to the control treatment (19.29±2.35). Adult emergence rates were significantly high in the control treatments (57.50±2.66%) compared to I. javanica (15.00±1.47%) and P. lilacinum (28.75±4.78%) treatments. This study provides evidence that endophytic isolates of I. javanica and P. lilacinum have a biocontrol potentials for used against whiteflies and could also explored as plant growth promoters.

Matched MeSH terms: Hypocreales*
Fulltext Morphological and phylogenetic analysis of Fusarium solani species complex in Malaysia

Chehri K, Salleh B, Zakaria L

Microb Ecol, 2015 Apr;69(3):457-71.
PMID: 25238930 DOI: 10.1007/s00248-014-0494-2

Members of Fusarium solani species complex (FSSC) have been known as plant, animal, and human pathogens. Nevertheless, the taxonomic status of such an important group of fungi is still very confusing and many new species as well as lineages have been elucidated recently. Unfortunately, most of the new taxa came from temperate and subtropical regions. Therefore, the objectives of the present study were to identify strains of FSSC recovered from different sources in Malaysia. In the present study, 55 strains belonging to the FSSC were examined and phylogenetically analyzed on the basis of internal transcribed spacer (ITS) regions and partial translation elongation factor-1 (TEF-1α) sequences. Based on morphological features, a total of 55 strains were selected for molecular studies. Based on morphological features, the strains were classified into four described Fusarium species, namely Fusarium keratoplasticum, Fusarium falciforme, FSSC 5, and Fusarium cf. ensiforme, and one unknown phylogenetic species was introduced. Although the data obtained from morphological and molecular studies sufficiently supported each other, the phylogenetic trees based on ITS and TEF-1α dataset clearly distinguished closely related species and distinctly separated all morphological taxa. All members of FSSC in this research were reported for the first time for Malaysian mycoflora.

Matched MeSH terms: Hypocreales/cytology*; Hypocreales/genetics*; Hypocreales/metabolism
Enhancing photosynthesis and root development for better fruit quality, aroma, and lessening of radioactive materials in key lime (Citrus aurantifolia) using Trichoderma harzianum and Bacillus thuringiensis

Salem A, Khandaker MM, Mahmud K, Alsufyani SJ, Majrashi AA, Rashid ZM, et al.

Plant Physiol Biochem, 2024 Jan;206:108295.
PMID: 38154296 DOI: 10.1016/j.plaphy.2023.108295

The present study was conducted to investigate the effects of Trichoderma harzianum and Bacillus thuringiensis alone or with gradual levels of NPK on photosynthesis, growth, fruit quality, aroma improvement and reduced radionuclides of key lime fruits. The lemon seedlings were treated with (T0) without fertilizers as control, (T1) 100g of NPK at 100%, (T2) 5 g of Trichoderma. harzianum at 50% + 50 g of NPK at 50%, (T3) 5 g of Bacillus thuringiensis at 50% + 50 g of NPK at 50 %, (T4) 7.5 g of Trichoderma harzianum at 75% + 25 g of NPK at 25 %, (T5) 7.5 g of Bacillus thuringiensis at 75% + 25 g of NPK at 25 %, (T6) 10 g of Trichoderma harzianum at 100 % and (T7)10 g of Bacillus thuringiensis at 100 %. The results showed that T2 increased net photosynthetic rate, stomatal conductance, transpiration rate, internal CO2 concentration, fresh and dry root biomass by 209%, 74%, 56%, 376%, 69.4% and 71.6%, while, T5 increased root volume, root length, and root tip number by 27.1%, 167%, and 67%, respectively over the control trees. The microbial treatments developed cortex, vascular cylinder and tracheal elements of the root. Fruit number, length, diameter, weight, pulp thickness, pulp/peel ratio, juice, total soluble solids (TSS), pigment contents and antioxidant activity increased significantly in the T2 treatment. Vitamin C, total phenols, total flavonoids, and total sugar content increased by 1.59-, 1.66-, 1.44- and 2.07- fold in T5 treated fruits compared to the control. The two microbes increased volatile compounds and decreased radionucleotides in the fruit, moreover, 27 identified and 2 (two) unmatched volatile compounds were identified by GCMS analysis. It is concluded that T. harzianum and B. thuringiensis with 25-50 g NPK treatments improved photosynthesis, root structure, fruit growth, fruit quality, aroma and lessened radionuclides in key lime fruits.

Matched MeSH terms: Hypocreales*
Fulltext Agro-ecological variations of sheath rot disease of rice caused by Sarocladium oryzae and DNA fingerprinting of the pathogen's population structure

Tajul Islam Chowdhury M, Salim Mian M, Taher Mia MA, Rafii MY, Latif MA

Genet. Mol. Res., 2015 Dec 28;14(4):18140-52.
PMID: 26782461 DOI: 10.4238/2015.December.23.1

To examine the impact of regional and seasonal variations on the incidence and severity of sheath rot, a major seed-borne disease of rice caused by Sarocladium oryzae, data on incidence and severity were collected from 27 selected fields in the Gazipur, Rangpur, Bogra, Chittagong, Comilla, Gopalgonj, Jessore, Manikgonj, and Bhola districts of Bangladesh in rain-fed and irrigated conditions. Cultural variability of 29 pathogen isolates obtained from 8 different locations was studied on potato dextrose agar (PDA) and genetic variability was determined by DNA fingerprinting using variable number tandem repeat-polymerase chain reaction markers. Overall, disease incidence and severity were higher in irrigated rice. Disease incidence and severity were highest in the Bhola district in rain-fed rice and lowest in irrigated rice. Mycelial growth of 29 representative isolates was found to vary on PDA and the isolates were divided into 6 groups. The range of the overall size of conidia of the selected isolates was 2.40-7.20 x 1.20-2.40 μm. Analysis of the DNA fingerprint types of the 29 isolates of S. oryzae, obtained from the amplification reactions, revealed 10 fingerprinting types (FPTs) that were 80% similar. FPT-1 was the largest group and included 13 isolates (44.8%), while FPT-2 was the third largest group and included 3 isolates. Each of FPT-3, 4, 5, and 6 included only 1 isolate. We observed no relationship between cultural and genetic groupings.

Matched MeSH terms: Hypocreales/genetics*; Hypocreales/pathogenicity
Molecular docking and molecular dynamics simulations studies on β-glucosidase and xylanase Trichoderma asperellum to predict degradation order of cellulosic components in oil palm leaves for nanocellulose preparation

Bahaman AH, Wahab RA, Abdul Hamid AA, Abd Halim KB, Kaya Y

J Biomol Struct Dyn, 2021 Apr;39(7):2628-2641.
PMID: 32248752 DOI: 10.1080/07391102.2020.1751713

Literature has shown that oil palm leaves (OPL) can be transformed into nanocellulose (NC) by fungal lignocellulosic enzymes, particularly those produced by the Trichoderma species. However, mechanism of β-glucosidase and xylanase selectivity to degrade lignin, hemicellulose and cellulose in OPL for NC production remains relatively vague. The study aimed to comprehend this aspect by an in silico approach of molecular docking, molecular dynamics (MD) simulation and Molecular-mechanics Poisson-Boltzmann surface area (MM-PBSA) analysis, to compare interactions between the β-glucosidase- and xylanase from Trichoderma asperellum UC1 in complex with each substrate. Molecular docking of the enzyme-substrate complex showed residues Glu165-Asp226-Glu423 and Arg155-Glu210-Ser160 being the likely catalytic residues of β-glucosidase and xylanase, respectively. The binding affinity of β-glucosidase for the substrates are as follows: cellulose (-8.1 kcal mol-1) > lignin (-7.9 kcal mol-1) > hemicellulose (-7.8 kcal mol-1), whereas, xylanase showed a corresponding preference for; hemicellulose (-6.7 kcal mol-1) > cellulose (-5.8 kcal mol-1) > lignin (-5.7 kcal mol-1). Selectivity of both enzymes was reiterated by MD simulations where interactions between β-glucosidase-cellulose and xylanase-hemicellulose were the strongest. Notably low free-binding energy (ΔGbind) of β-glucosidase and xylanase in complex with cellulose (-207.23 +/- 47.13 kJ/mol) and hemicellulose (-131.48 +/- 24.57 kJ/mol) were observed, respectively. The findings thus successfully identified the cellulose component selectivity of the polymer-acting β-glucosidase and xylanase of T. asperellum UC1.Communicated by Ramaswamy H. Sarma.

Matched MeSH terms: Hypocreales
A novel study based on adaptive metal tolerance behavior in fungi and SEM-EDX analysis

Chen SH, Ng SL, Cheow YL, Ting ASY

J Hazard Mater, 2017 Jul 15;334:132-141.
PMID: 28407540 DOI: 10.1016/j.jhazmat.2017.04.004

Four fungal isolates: Simplicillium chinense (iso 9, accession no. KX425621), Penicillium simplicissimum (iso 10, KP713758), Trichoderma asperellum (iso 11, KP792512), and Coriolopsis sp. (1c3, KM403574) were subjected to a series of induced-tolerance training under high metal concentrations to determine if greater tolerance could be achieved from constant exposure to such conditions. Adaptive tolerance assay (Tolerance Index, TI) and Field-Emission Scanning Electron Microscopy with Energy Dispersive X-ray (SEM-EDX) characterized their metal tolerance. "Untrained" S. chinense, P. simplicissimum and T. asperellum showed tolerance towards 4000-4500ppm Al(III) (TI: 0.64-0.71), 1000ppm Cr(III) (0.52-0.83) and Pb(II) (0.32-0.88). With tolerance training, tolerance towards 2000-6000ppm Al(III), 500-3000ppm Pb(II) and 2000-3000ppm Cr(III) were achieved (TI: 0.01-0.82) compared to untrained cultures (0.00-0.59). In contrast, tolerance training for Coriolopsis sp. and P. simplicissimum was less successful, with TI values similar or lower than untrained cultures. SEM-EDX analysis proposed biosorption and bioaccumulation as mechanisms for metal removal. The latter was demonstrated with the removal of Cr(III) and Pb(II) by S. chinense (12.37 and 11.52mgg-1, respectively) and T. asperellum (10.44 and 7.50mgg-1). Induced-tolerance training may render benefit in the long run, but this delicate approach is suggestively species and metal dependent.

Matched MeSH terms: Hypocreales/drug effects*; Hypocreales/physiology; Hypocreales/ultrastructure
Investigating process parameters to enhance (hemi)cellulolytic enzymes activity produced by Trichoderma reesei RUT-C30 using deoiled oil palm mesocarp fiber in solid-state fermentation

Al-Qassab AA, Zakaria MR, Yunus R, Salleh MAM, Mokhtar MN

Int J Biol Macromol, 2024 Sep;276(Pt 2):134030.
PMID: 39038578 DOI: 10.1016/j.ijbiomac.2024.134030

This study investigates the synthesis of (hemi)cellulolytic enzymes, including endoglucanase (CMCase), xylanase, and β-glucosidase, employing Trichoderma reesei RUT-C30 and deoiled oil palm mesocarp fiber (OPMF) through solid-state fermentation (SSF). The objective was to determine the optimal process conditions for achieving high enzyme activities through a one-factor-at-a-time approach. The study primarily focused on the impact of the solid-to-liquid ratio, incubation period, initial pH, and temperature on enzyme activity. The effects of OPMF pretreatment, particularly deoiling and fortification, were explored. This approach significantly improved enzyme activity levels compared to the initial conditions, with CMCase increasing by 111.6 %, xylanase by 665.2 %, and β-Glucosidase by 1678.1 %. Xylanase and β-glucosidase activities, peaking at 1346.75 and 9.89 IU per gram dry substrate (GDS), respectively, under optimized conditions (1:4 ratio, pH 7.5, 20 °C, 9-day incubation). With lower moisture levels, CMCase reached its maximum activity of 227.84 IU/GDS. The study highlights how important it is for agro-industrial byproducts to support environmentally sustainable practices in the palm oil industry. It also emphasizes how differently each enzyme reacts to changes in process parameters.

Matched MeSH terms: Hypocreales/enzymology
Classification of the guava wilt fungus Myxosporium psidii, the palm pathogen Gliocladium vermoesenii and the persimmon wilt fungus Acremonium diospyri in Nalanthamala

Schroers HJ, Geldenhuis MM, Wingfield MJ, Schoeman MH, Yen YF, Shen WC, et al.

Mycologia, 2005 Mar-Apr;97(2):375-95.
PMID: 16396346

Psidium guajava wilt is known from South Africa, Malaysia and Taiwan. The fungus causing this disease, Myxosporium psidii, forms dry chains of conidia on surfaces of pseudoparenchymatous sporodochia, which develop in blisters on bark. Similar sporodochia are characteristic of Nalanthamala madreeya, the type species of Nalanthamala. Nalanthamala, therefore, is the appropriate anamorph genus for Myxosporium psidii, while Myxosporium is a nomen nudum (based on M. croceum). For M. psidii the combination Nalanthamala psidii is proposed. Nalanthamala psidii, the palm pathogen Gliocladium (Penicillium) vermoesenii, another undescribed anamorphic species from palm, two species of Rubrinectria and the persimmon pathogen Acremonium diospyri are monophyletic and belong to the Nectriaceae (Hypocreales) based on partial nuclear large subunit ribosomal DNA (LSU rDNA) analyses. Rubrinectria, therefore, is the teleomorph of Nalanthamala, in which the anamorphs are classified as N. vermoesenii, N. diospyri or Nalanthamala sp. Nalanthamala squamicola, the only other Nalanthamala species, has affinities with the Bionectriaceae and is excluded from this group. Rubrinectria/Nalanthamala species form dimorphic conidiophores and conidia in culture. Fusiform, cylindrical, or allantoid conidia arise in colorless liquid heads on acremonium-like conidiophores; ovoidal conidia with somewhat truncated ends arise in long, persistent, dry chains on penicillate conidiophores. No penicillate but irregularly branched conidiophores were observed in N. diospyri. Conidia of N. psidii that are held in chains are shorter than those of N. madreeya, of which no living material is available. Nalanthamala psidii and N. diospyri are pathogenic specifically to their hosts. They form pale yellow to pale orange or brownish orange colonies, respectively, and more or less white conidial masses. Most strains of Rubrinectria sp., Nalanthamala sp. and N. vermoesenii originate from palm hosts, form mostly greenish or olive-brown colonies and white-to-salmon conidial masses. They form a monophyletic clade to which Nalanthamala psidii and N. diospyri are related based on analyses of the internal transcribed spacer regions and 5.8S rDNA (ITS rDNA), LSU rDNA, and partial beta-tubulin gene. Few polymorphic sites in the ITS rDNA and beta-tubulin gene indicate that Nalanthamala psidii comprises two lineages, one of which has been detected only in South Africa.

Matched MeSH terms: Hypocreales/classification*; Hypocreales/cytology; Hypocreales/genetics; Hypocreales/isolation & purification*
Paecilomyces lilacinus fungaemia in an AIDS patient: the importance of mycological diagnosis

Ding CH, Tzar MN, Rahman MM, Muttaqillah NA, Redzuan SR, Periyasamy P

Pak J Med Sci, 2014 Jul;30(4):914-6.
PMID: 25097544

Fungaemia due to Paecilomyces lilacinus is generally not considered in AIDS patients because this condition is not categorised as an AIDS-indicator illness. We report a case of a 25-year-old lady who presented to our hospital with Guillain-Barré Syndrome, with the subsequent development of refractory fungaemia, multi-organ failure and disseminated intravascular coagulopathy. Amphotericin B was given as empirical antifungal therapy. HIV screening was reactive and Paecilomyces lilacinus was isolated from her blood. The fungaemia did not resolve after one week of amphotericin B treatment. The addition of itraconazole was also unsuccessful in clearing the fungaemia. Accurate mycological diagnosis is important in the care of AIDS patients with fungaemia because of the risk of treatment failure with empirical therapy.

Matched MeSH terms: Hypocreales
Fulltext Evidence to support cultivated fruiting body of Ophiocordyceps sinensis (Ascomycota)'s role in relaxing airway smooth muscle

Booi HN, Pang LY, Lee MK, Fung SY, Ng CL, Ng ST, et al.

J Ethnopharmacol, 2025 Jan 10;336:118727.
PMID: 39182700 DOI: 10.1016/j.jep.2024.118727

ETHNOPHARMACOLOGICAL RELEVANCE: Ophiocordyceps sinensis (O. sinensis) is a genus of Ascomycete fungus that is endemic to the alpine meadows of the Tibetan Plateau and adjoining Himalayas. It has been used traditionally as a tonic to improve respiratory health in ancient China as well as to promote vitality and longevity. Bioactive components found in O. sinensis such as adenosine, cordycepin, 3-deoxyadenosine, L-arginine and polysaccharides have gained increasing interest in recent years due to their antioxidative and other properties, which include anti-asthmatic, antiviral, immunomodulation and improvement of general health.
AIM OF THE STUDY: This study's primary aim was to investigate the effect of a cultivated fruiting body of O. sinensis strain (OCS02®) on airways patency and the secondary focus was to investigate its effect on the lifespan of Caenorhabditis elegans.
MATERIALS AND METHODS: A cultivated strain, OCS02®, was employed and the metabolic profile of its cold-water extract (CWE) was analysed through liquid chromatography-mass spectrometry (LC-MS). Organ bath approach was used to investigate the pharmacological properties of OCS02® CWE when applied on airway tissues obtained from adult male Sprague-Dawley rats. The airway relaxation mechanisms of OCS02® CWE were explored using pharmacological tools, where the key regulators in airway relaxation and constriction were investigated. For the longevity study, age-synchronised, pos-1 RNAi-treated wild-type type Caenorhabditis elegans at the L4 stage were utilised for a lifespan assay.
RESULTS: Various glycopeptides and amino acids, particularly a high concentration of L-arginine, were identified from the LC-MS analysis. In airway tissues, OCS02® CWE induced a significantly greater concentration-dependent relaxation when compared to salbutamol. The relaxation response was significantly attenuated in the presence of NG-Nitro-L-arginine methyl ester (L-NAME), 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one (ODQ) and several K+ channel blockers. The longevity effect induced by OCS02® CWE (5 mg/mL and above) was observed in C. elegans by at least 17%.
CONCLUSIONS: These findings suggest that the airway relaxation mechanisms of OCS02® CWE involved cGMP-dependent and cGMP-independent nitric oxide signalling pathways. This study provides evidence that the cultivated strain of OCS02® exhibits airway relaxation effects which supports the traditional use of its wild O. sinensis in strengthening respiratory health.

Matched MeSH terms: Hypocreales
Medroxyprogesterone derivatives from microbial transformation as anti-proliferative agents and acetylcholineterase inhibitors (combined in vitro and in silico approaches)

Yusop SNW, Imran S, Adenan MI, Sultan S

Steroids, 2020 12;164:108735.
PMID: 32976918 DOI: 10.1016/j.steroids.2020.108735

The fungal transformations of medroxyrogesterone (1) were investigated for the first time using Cunninghamella elegans, Trichothecium roseum, and Mucor plumbeus. The metabolites obtained are as following: 6β, 20-dihydroxymedroxyprogesterone (2), 12β-hydroxymedroxyprogesterone (3), 6β, 11β-dihydroxymedroxyprogesterone (4), 16β-hydroxymedroxyprogesterone (5), 11α, 17-dihydroxy-6α-methylpregn-4-ene-3, 20-dione (6), 11-oxo-medroxyprogesterone (7), 6α-methyl-17α-hydroxypregn-1,4-diene-3,20-dione (8), and 6β-hydroxymedroxyprogesterone (9), 15β-hydroxymedroxyprogesterone (10), 6α-methyl-17α, 11β-dihydroxy-5α-pregnan-3, 20-dione (11), 11β-hydroxymedroxyprogesterone (12), and 11α, 20-dihydroxymedroxyprogesterone (13). Among all the microbial transformed products, the newly isolated biotransformed product 13 showed the most potent activity against proliferation of SH-SY5Y cells. Compounds 12, 5, 6, 9, 11, and 3 (in descending order of activity) also showed some extent of activity against SH-SY5Y tumour cell line. The never been reported biotransformed product, 2, showed the most potent inhibitory activity against acetylcholinesterase. Molecular modelling studies were carried out to understand the observed experimental activities, and also to obtain more information on the binding mode and the interactions between the biotransformed products, and enzyme.

Matched MeSH terms: Hypocreales/metabolism
Fulltext Multicomponent carbohydrase system from Trichoderma reesei: A toolbox to address complexity of cell walls of plant substrates in animal feed

Rangel Pedersen N, Tovborg M, Soleimani Farjam A, Della Pia EA

PLoS One, 2021;16(6):e0251556.
PMID: 34086701 DOI: 10.1371/journal.pone.0251556

A diverse range of monocot and dicot grains and their by-products are commonly used in the animal feed industry. They all come with complex and variable cell wall structures which in turn contribute significant fiber to the complete feed. The cell wall is a highly interconnected matrix of various polysaccharides, proteins and lignin and, as such, requires a collaborative effort of different enzymes for its degradation. In this regard, we investigated the potential of a commercial multicomponent carbohydrase product from a wild type fermentation of Trichoderma reesei (T. reesei) (RONOZYME® MultiGrain) in degrading cell wall components of wheat, barley, rye, de-oiled rice bran, sunflower, rapeseed and cassava. A total of thirty-one different enzyme proteins were identified in the T. Reesei carbohydrase product using liquid chromatography with tandem mass spectrometry LC-MS/MS including glycosyl hydrolases and carbohydrate esterases. As measured by in vitro incubations and non-starch polysaccharide component analysis, and visualization by immunocytochemistry and confocal microscopy imaging of immuno-labeled samples with confocal microscopy, the carbohydrase product effectively solubilized cellulolytic and hemicellulolytic polysaccharides present in the cell walls of all the feed ingredients evaluated. The T. reesei fermentation also decreased viscosity of arabinoxylan, xyloglucan, galactomannan and β-glucan substrates. Combination of several debranching enzymes including arabinofuranosidase, xylosidase, α-galactosidase, acetyl xylan esterase, and 4-O-methyl-glucuronoyl methylesterase with both GH10 and GH11 xylanases in the carbohydrase product resulted in effective hydrolyzation of heavily branched glucuronoarabinoxylans. The different β-glucanases (both endo-β-1,3(4)-glucanase and endo-β-1,3-glucanase), cellulases and a β-glucosidase in the T. reesei fermentation effectively reduced polymerization of both β-glucans and cellulose polysaccharides of viscous cereals grains (wheat, barley, rye and oat). Interestingly, the secretome of T. reesei contained significant amounts of an exceptional direct chain-cutting enzyme from the GH74 family (Cel74A, xyloglucan-specific β-1,4-endoglucanase), that strictly cleaves the xyloglucan backbone at the substituted regions. Here, we demonstrated that the balance of enzymes present in the T. reesei secretome is capable of degrading various cell wall components in both monocot and dicot plant raw material used as animal feed.

Matched MeSH terms: Hypocreales/metabolism*
Immunomodulatory Properties of Water-Soluble Polysaccharides Extracted from the Fruiting Body of Chinese Caterpillar Mushroom, Ophiocordyceps sinensis Cultivar OCS02® (Ascomycetes)

Yap ACS, Li X, Yap YHY, Razif MFM, Jamil AHA, Ng ST, et al.

Int J Med Mushrooms, 2020;22(10):967-977.
PMID: 33426826 DOI: 10.1615/IntJMedMushrooms.2020036351

Ophiocordyceps sinensis (=Cordyceps sinensis) has been known for its various medicinal properties, in particular immunomodulatory activities associated with its polysaccharides. In this study, the fruiting body of O. sinensis cultivar OCS02® was investigated for its chemical composition and monosaccharide profile. Cold water extract (CWE) obtained from this fruiting body was fractionated by molecular weight (MW) into high (HMW), medium (MMW), and low (LMW) fractions. Polysaccharides in the extract and fractions were identified as heteroglycans containing mostly glucose and mannose with small amounts of galactose, fucose, arabinose, and xylose. The immunomodulatory potential of these heteroglycans was evaluated by induction of cytokine/chemokine secretion using murine macrophage RAW 264.7. All treatments showed significant modulation of IL-6, IL-9, MIP-2, and TIMP-1, especially for CWE, HMW, and MMW, which might be due to their high ratios of glucose and the presence of protein. Further investigation on the structure-function relationship of these fruiting body polysaccharide fractions is needed to delineate the underlying mechanism of their immunomodulatory effect both in vitro and in vivo.

Matched MeSH terms: Hypocreales/chemistry*

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