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.

Dietary carbon sources in headwater stream food webs are divided into allochthonous and autochthonous organic matters. We hypothesized that: 1) the dietary allochthonous contribution for fish in headwater stream food webs positively relate with canopy cover; and 2) essential fatty acids originate from autochthonous organic matter regardless of canopy covers, because essential fatty acids, such as 20:5ω3 and 22:6ω3, are normally absent in allochthonous organic matters. We investigated predatory fish Salvelinus leucomaenis stomach contents in four headwater stream systems, which are located in subarctic region in northern Japan. In addition, stable carbon and nitrogen isotope ratios, fatty acid profile, and stable carbon isotope ratios of essential fatty acids were analyzed. Bulk stable carbon analysis showed the major contribution of autochthonous sources to assimilated carbon in S. leucomaenis. Surface baits in the stomach had intermediate stable carbon isotope ratios between autochthonous and allochthonous organic matter, indicating aquatic carbon was partly assimilated by surface baits. Stable carbon isotope ratios of essential fatty acids showed a positive relationship between autochthonous sources and S. leucomaenis across four study sites. This study demonstrated that the main supplier of dietary carbon and essential fatty acids was autochthonous organic matter even in headwater stream ecosystems under high canopy cover.

Mn-rich filamentous particles present in stratified water environments are considered bacteriogenic; however, little is known about their causative agents. This study investigated the production of these particles by an alphaproteobacterium, Bosea sp. strain BIWAKO-01. Particle formation was promoted in static cultures with slightly viscous medium at pH 6.0−6.3 under low-O₂ conditions. The Mn(II) oxidation in cultures was slower in higher O₂ concentration. These results suggested that pH and O₂ concentration are important factors affecting filamentous Mn particle formation in the Mn(II) oxidizer. Lectin staining followed by fluorescence microscopy revealed the presence of specific carbohydrates in the filamentous structures. In addition, transmission electron microscopy, high angle annular dark field scanning transmission electron microscopy, and electron energy loss spectroscopy revealed the structural and spatial associations of Mn with O and C on a nanometer scale in filaments. The results suggested the occurrence of sheet-type Mn oxide likely due to the catalytic activity in exopolymeric substances including acidic polysaccharides.

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