Optimizing the tray dryer temperature and time of white corn flour culture

This research aimed to optimize the tray dryer temperature and time of white corn flour culture by Response Surface Methodology (RSM). There were two cultures used in this research, namely Amylolytic Culture (AC) and Complete Culture (CC). AC consisted of Penicillium citrinum, Aspergillus niger, Acremonium strictum, and Candida famata, while CC consisted of Penicillium chrysogenum, Penicillium citrinum, Aspergillus niger, Rhizopus stolonifer, Rhizopus oryzae, Fusarium oxysporum, Acremonium strictum, Candida famata, Kodamaea ohmeri and Candida krusei/incospicua. The independent variables in this study were drying temperature and time, where the quality indicators used were total viability of mold and yeast, water content, water activity, and pH. This research used a factor response surface methodology. Data were analyzed by ANOVA with an α level of 95%. The result of this research showed that the optimum drying process for AC starter was 40°C for 10 hrs, with characteristic response viability 8.8×107 CFU/g, water activity 0.43, water content 8.90%, and pH 4.05. CC starter showed an optimum drying process at 49°C for 4.5 hrs, with characteristic response viability 4.9×107 CFU/g, water activity 0.49, water content 7.02%, and pH 3.95. The optimum tray dryer temperatures and times were achieved for AC and CC starters.

Preferential Elimination of Ba2+ through Irreversible Biogenic Manganese Oxide Sequestration

Biogenic manganese oxides (BMOs) formed in a culture of the Mn(II)-oxidizing fungus Acremonium strictum strain KR21-2 are known to retain enzymatic Mn(II) oxidation activity. Consequently, these are increasingly attracting attention as a substrate for eliminating toxic elements from contaminated wastewaters. In this study, we examined the Ba2+ sequestration potential of enzymatically active BMOs with and without exogenous Mn2+. The BMOs readily oxidized exogenous Mn2+ to produce another BMO phase, and subsequently sequestered Ba2+ at a pH of 7.0, with irreversible Ba2+ sequestration as the dominant pathway. Extended X-ray absorption fine structure spectroscopy and X-ray diffraction analyses demonstrated alteration from turbostratic to tightly stacked birnessite through possible Ba2+ incorporation into the interlayer. The irreversible sequestration of Sr2+, Ca2+, and Mg2+ was insignificant, and the turbostratic birnessite structure was preserved. Results from competitive sequestration experiments revealed that the BMOs favored Ba2+ over Sr2+, Ca2+, and Mg2+. These results explain the preferential accumulation of Ba2+ in natural Mn oxide phases produced by microbes under circumneutral environmental conditions. These findings highlight the potential for applying enzymatically active BMOs for eliminating Ba2+ from contaminated wastewaters.

Stereochemical Characterisation of the Non-Canonical α-Humulene Synthase from Acremonium strictum

The non-canonical fungal α-humulene synthase was investigated through isotopic labelling experiments for its stereochemical course regarding inversion or retention at C-1, the face selectivity at C-11, and the stereoselectivity of...

Bacterial wetwood of silver birch (Betula pendula roth): symptomology, etiology and pathogenesis

The article is focused on microbiological and silvicultural properties of bacterial wetwood of silver birch (Betula pendula), also known as European white birch. During the active phase of the disease, bacterial wetwood (i.e. bacterial dropsy, vascular parenchymatous bacteriosis or flux slime) is characterised by crust and periderm bloating, necrotic wet stains and abundance of exudate. The disease is more likely to occur in older (r = 0.56, p < 0.01) and less-dense (r = −0.29, p < 0.01) stands.The statistical model showed that the chance of bacterial wetwood increases with birch age by 0.36% per year. The stands with birch proportion of over 70% demonstrated 15.3% lower infection rate compared to the stands with lower birch presence. The stands with lower stocking demonstrated a higher proportion of infected tree distribution by 7.5% compared to the stands with higher birch representation. The most vulnerable were larger, older B. pendula trees with longitudinally fissured bark that grow on poorer soils and experience frequent water stress.Birch associations with Pteridium aquilinum and Vaccinium myrtillus were more susceptible to infection (31.6% and 44.3%, respectively), whereas associations with Brachypodium sylvaticum, Sphagnum palustre and Calluna vulgaris were at lower risk. Strong ecological and trophic association of bacterial wetwood was present between silver birch and Tremex spp., particularly Tremex fuscicornis. Mycobiota was represented by Rhizopus microsporus, Mucor mucedo, Penicillium aurantiogriseum, Penicillium purpurogenum and Acremonium strictum.Enterobacter, Xanthomonas, Pantoea and Bacillus spp. associated with bacterial wetwood of silver birch were isolated. Enterobacter nimipressuralis was found to be the primary causative agent through means of artificial infection, while other bacteria were found to be either weak pathogens or concomitant. E. nimipressuralis formed the largest number of colony-forming units (CFU) for bark and cambium (164 and 127 CFU, respectively) and was also found in a small amount as a vital obligate in the automicrobiota in healthy birch trees.

ANTAGONISTIC ACTIVITY OF A NEW STRAIN OF TRICHODERMA VIRIDE AND ITS EFFECT ON MICROMYCETES POPULATIONS IN THE ROOT ZONE OF CORN PLANTS

Goal. To investigate the antagonistic activity of a new strain of Trichoderma viride F-100076and its effect on the formation of micromycetes populations in the root zone of corn plants underfield conditions. Methods. The antagonistic activity of T. viride F-100076 was studied by the method of mixed (counter) cultures on wort agar using phytopathogenic fungi, which were isolated andidentified in the Laboratory of Plant-Microbial Interactions. The appearance and type of relationship were registered using a scale modified by Symonian and Mamikonian. The number of micromycetes was determined by the method of soil dilutions. Isolation, accounting and cultivation of fungiwas carried out according to conventional methods. Micromycetes were identified according to thedeterminants appropriate for a specific systematic group of micromycetes. Results. It was foundthat T. viride IMB F-100076 is characterized by high antagonistic activity against a wide range ofphytopathogenic fungi, showing hyperparasitism as early as on the eighth day. The highest antagonistic activity of the strain was found against: Alternaria radicina, Acremonium strictum, Acremonium сucurbitacearum, Fusarium oxysporum var. orthoceras, Fusarium moniliforme var. lactis, Torula expansa (5 points on the corresponding Symonian and Mamikonian scale). Data from the mycological analysis of the sod-podzolic soil of the corn rhizosphere showed that the mycocenosis ofthe sod-podzolic soil of the corn rhizosphere was formed by micromycetes belonging to the generaAcremonium Link, Cladosporium Corda, Fusarium Link:Fr, Gliocladium Corda, Mucor Mich, Penicillium Link:Fr, Rhizopus Ehrenb, Trichoderma Hers, among which the most represented were micromycetes of the genus Penicillium (59 %). The total number of fungi in the control variant was291.00 ± 79.67 thousand CFU/g of soil. The introduction of straw affected both the total number ofmicromycetes and the genus composition of fungi. The total number of fungi in the variant withstraw increased 2.6 times and amounted to 744.00 ± 114.67 thousand CFU/g of soil. The number ofrepresentatives of all studied genera of micromycetes also increased. In addition, the introductionof straw provoked the development of fungi of Bipolaris and Fusarium genera, which can be considered a negative outcome since representatives of these species are commonly recognised as pathogens of root diseases. Application of the fungus antagonist T. viride IMB F-100076 to the soilalong with straw did not significantly affect the total number of micromycetes. At the same time, a displacement of fungi of the genus Bipolaris and Fusarium from the rhizosphere of corn was registered.The number of fusaria decreased from 96.00 ± 5.44 to 23.00 ± 2.32 thousand CFU/g of soil or almost4 times and reached the level of the control variant. Fungi of the genus Bipolaris in the variant withthe introduction of trichoderma were not detected. Conclusion. The antagonist fungus T. virideF-100076, introduced into the soil along with straw, strikes root in the soil and exhibits antagonisticactivity against micromycetes of the genera Bipolaris and Fusarium, which are commonly represented by root rot pathogens of many crops. Thus, the new strain T. viride F-100076 allows increasing theantagonistic potential of the rhizosphere soil of corn and protecting plants from pathogens.

Molecular Cloning and Heterologous Expression of Manganese(II)-Oxidizing Enzyme from Acremonium strictum Strain KR21-2

Diverse ascomycete fungi oxidize manganese(II) [Mn(II)] and produce Mn(III, IV) oxides in terrestrial and freshwater environments. Although multicopper oxidase (MCO) is considered to be a key catalyst in mediating Mn(II) oxidation in ascomycetes, the responsible gene and its product have not been identified. In this study, a gene, named mco1, encoding Mn(II)-oxidizing MCO from Acremonium strictum strain KR21-2 was cloned and heterologously expressed in the methylotrophic yeast Pichia pastoris. Based on the phylogenetic relationship, similarity of putative copper-binding motifs, and homology modeling, the gene product Mco1 was assigned to a bilirubin oxidase. Mature Mco1 was predicted to be composed of 565 amino acids with a molecular mass of 64.0 kDa. The recombinant enzyme oxidized Mn(II) to yield spherical Mn oxides, several micrometers in diameter. Zinc(II) ions added to the reaction mixture were incorporated by the Mn oxides at a Zn/Mn molar ratio of 0.36. The results suggested that Mco1 facilitates the growth of the micrometer-sized Mn oxides and affects metal sequestration through Mn(II) oxidation. This is the first report on heterologous expression and identification of the Mn(II) oxidase enzyme in Mn(II)-oxidizing ascomycetes. The cell-free, homogenous catalytic system with recombinant Mco1 could be useful for understanding Mn biomineralization by ascomycetes and the sequestration of metal ions in the environment

Sequestration and Oxidation of Cr(III) by Fungal Mn Oxides with Mn(II) Oxidizing Activity

Biogenic manganese oxides (BMOs) have gained increasing attention for environmental application because of their sequestration and oxidizing abilities for various elements. Oxidation and sequestration of Cr(III) by BMOs, however, still remain unknown. We prepared BMOs in liquid cultures of Acremonium strictum strain KR21-2, and subsequently conducted single or repeated treatment experiments in Cr(NO3)3 at pH 6.0. Under aerobic conditions, newly formed BMOs exhibited a rapid production of Cr(VI) without a significant release of Mn(II), demonstrating that newly formed BMO mediates a catalytic oxidation of Cr(III) with a self-regeneration step of reduced Mn. In anaerobic solution, newly formed BMOs showed a cessation of Cr(III) oxidation in the early stage of the reaction, and subsequently had a much smaller Cr(VI) production with significant release of reduced Mn(II). Extraordinary sequestration of Cr(III) was observed during the repeated treatments under anaerobic conditions. Anaerobically sequestered Cr(III) was readily converted to Cr(VI) when the conditions became aerobic, which suggests that the surface passivation is responsible for the anaerobic cessation of Cr(III) oxidation. The results presented herein increase our understanding of the roles of BMO in Cr(III) oxidation and sequestration processes in potential application of BMOs towards the remediation of Cr(III)/Cr(VI) in contaminated sites.

Optimization of temperature and drying time of indigenous cocktail yeast mold culture using response surface methodology (RSM)

The study was conducted to obtain an optimal combination of time and temperature of the drying process of indigenous cocktail yeast mold culture using RSM. The cocktail yeast mold culture was dried using an oven. The cocktail cultures contain Penicillium citrinum, Aspergillus niger, Acremonium strictum, and Candida famata, namely AC (Amylolytic Culture). The Response Surface Methods (RSM) with Design-Expert® 7.00 software, namely Mixture design with D-optimal was performed. The drying time was between 24- 48 hrs and the drying temperature was between 40-50oC. The total of 16 formulas of the combination of drying time and temperature for processing the dried cultures were produced by RSM. The response chosen was total viability of mold and yeast, water content, water activity, and pH. The result of optimization and verification was obtained by the model: pH (AC) = -0.058A - 1.56 x 10-003B + 7.13, where A = drying temperature ( oC), B = drying time (hr). The AC optimization was achieved at a combination of drying temperatures and time of 50oC for 48 hrs. Desirability values were 0.729. The optimum formula for AC has viability of total yeast mold of 7.39 x 106 CFU/g, moisture content of 5.62%, aw 0.303, and pH 4.18.