Protein secretion and associated stress in industrially employed filamentous fungi.

Application of filamentous fungi for the production of commercial enzymes such as amylase, cellulase, or xylanase is on the rise due to the increasing demand to degrade several complex carbohydrates as raw material for biotechnological processes. Also, protein production by fungi for food and feed gains importance. In any case, the protein production involves both cellular synthesis and secretion outside of the cell. Unfortunately, the secretion of proteins or enzymes can be hampered due to accumulation of unfolded or misfolded proteins in the endoplasmic reticulum (ER) as a result of too high synthesis of enzymes or (heterologous) protein expression. To cope with this ER stress, the cell generates a response known as unfolded protein response (UPR). Even though this mechanism should re-establish the protein homeostasis equivalent to a cell under non-stress conditions, the enzyme expression might still suffer from repression under secretory stress (RESS). Among eukaryotes, Saccharomyces cerevisiae is the only fungus, which is studied quite extensively to unravel the UPR pathway. Several homologs of the proteins involved in this signal transduction cascade are also found in filamentous fungi. Since RESS seems to be absent in S. cerevisiae and was only reported in Trichoderma reesei in the presence of folding and glycosylation inhibitors such as dithiothreitol and tunicamycin, more in-depth study about this mechanism, specifically in filamentous fungi, is the need of the hour. Hence, this review article gives an overview on both, protein secretion and associated stress responses in fungi. KEY POINTS: • Enzymes produced by filamentous fungi are crucial in industrial processes • UPR mechanism is conserved among many fungi, but mediated by different proteins • RESS is not fully understood or studied in industrially relevant filamentous fungi.

Body mass index and 12-year mortality among older Mexican Americans aged 75 years and older.

BACKGROUND: The role of obesity in mortality in the very old and old-oldest Hispanic population has not been studied. The objective of this study was to examine the effect of body mass index (BMI) on 12-year mortality among older Mexican Americans aged 75 years and older. METHODS: Twelve year prospective cohort study consisting of a population-based sample of 1415 non-institutionalized Mexican American men and women aged 75 and older from 5 southwestern states: Arizona, California, Colorado, New Mexico, and Texas. Data was from Wave 5 of the Hispanic Established Population for the Epidemiologic Study of the Elderly (2004/2005-2016). Socio-demographics, body mass index (BMI), self-reported medical conditions, disability, depressive symptoms, falls, Mini-Mental-State-Examination (MMSE), and Short Physical Performance Battery (SPPB) were assessed at baseline during 2004-2005. BMI (Kg/m2) was classified as underweight (< 18.5), normal weight (18.5 to < 25), overweight (25 to < 30), obesity category I (30 to < 35), and obesity category II/morbid obesity (≥ 35). For assessment of mortality, deaths were ascertained through the National Death Index and report from relatives. Cox proportional hazards regression analysis was performed to estimate the hazard ratio (HR) of 12-year mortality as a function of BMI categories at baseline. RESULTS: The mean BMI was 27.5 ± 1.7 with participants classified as 1.8% underweight, 30.8% normal weight, 39.2% overweight, 20.7% obesity category I, and 7.6% obesity category II/morbid obesity. Mexican Americans aged ≥75 years with overweight or obesity category I had a reduced HR of death (0.82, 95% CI = 0.70-0.96 and 0.75, 95% CI = 0.62-0.91, respectively) over 12-years of follow-up. The HR of death for underweight and obesity category II/morbid obesity participants was 1.59 (95% CI = 1.03-2.45) and 1.12 (95% CI = 0.85-1.46), respectively. Female participants and those with high scores in the MMSE and SPPB had decreased risk of death. CONCLUSIONS: This study showed the protective effect of overweight and obesity on mortality in Mexican Americans above 75 years of age, which might have implications when treating older adults with overweight and obesity.

Nectar Yeast Community of Tropical Flowering Plants and Assessment of Their Osmotolerance and Xylitol-Producing Potential.

Floral nectar is colonised by microbes, especially yeasts which alter the scent, temperature, and chemical composition of nectar, thereby playing an essential role in pollination. The yeast communities inhabiting the nectar of tropical flowers of India are not well explored. We isolated 48 yeast strains from seven different tropical flowering plants. Post MSP-PCR-based screening, 23 yeast isolates and two yeast-like fungi were identified, which belonged to 16 species of 12 genera viz. Candida (2 species), Aureobasidium (2 species), Metschnikowia (2 species), Meyerozyma (1 species), Saitozyma (1 species), Wickerhamomyces (1 species), Kodamaea (2 species), Pseudozyma (1 species), Starmerella (1 species), Hanseniaspora (1 species), Rhodosporidiobolus (1 species), Moesziomyces (1 species), and two putative novel species. All yeast strains were assessed for their osmotolerance abilities in high salt and sugar concentration. Among all the isolates, C. nivariensis (SRA2.2, SRA1.1 and SRA2.1), M. caribbica (SRA4.8 and SRA4.6), S. flava SRA4.2, and M. reukaufii SRA3.2 showed significant growth in high concentrations of sugar (40-50% glucose), as well as salt (12-15% NaCl). All 25 strains were also screened for their ability to utilise xylose to produce xylitol. Meyerozyma caribbica was the most efficient xylitol producer, wherein three strains of this species (SRA4.6, SRA4.1, and SRA4.8) generated 18.61 to 21.56 g l-1 of xylitol, with 0.465-0.539 g g-1 yields. Through this study, we draw attention towards the tropical floral nectar as a potential niche for the isolation of diverse, osmotolerant, and xylitol-producing yeasts. Such osmotolerant yeasts have potential applications in food industries and biofuel production.

A Rare Stinkhorn Fungus Itajahya rosea Attract Drosophila by Producing Chemical Attractants.

Itajahya rosea was found growing in association with Leucaena leucocephala plants at Savitribai Phule Pune University campus in India. The species identity was confirmed by phylogenetic analysis based on ITS and LSU regions of rDNA, wherein, our fugus was placed along with I. rosea in phylogenetic tree. It represents first record of I. rosea from India. Frequent visitation by Drosophila species on I. rosea fruiting body particularly on gleba was observed. The Drosophila got attracted to the detached gleba under the laboratory conditions and even sometimes, they prefer to sit over the gleba as compare to their food banana. It suggested that I. rosea gleba or pseudostipe produces some compounds for attraction and feeding behavior of Drosophila species. Therefore, we characterized the volatile attractants produced by gleba and pseudostipe of I. rosea by GC-MS analysis. Nineteen compounds were identified from gleba while nine compounds were recovered from the pseudostipe. Out of them, blends of three abundant odor producing volatile compounds were reported namely, Hexadecane, Pentadecane and Nonadecane, which are responsible for attraction of Drosophila toward the gleba. Three fatty acids namely 9,12-octadecadienoic acid (Z,Z), hexadecanoic acid and benzoic acid ethyl ester produced are served as an appetitive signal through olfactory response of Drosophila, so the flies were feed on the gleba. Two pheromones' compounds, heneicosane and (+)-(5S,9S)-5,9-dimethylpentadecane, were also reported in pseudostipe and gleba, respectively, which play a role in Drosophila for breeding. Our study highlights an intriguing chemical ecology of fungus-Drosophila interaction.

In Vitro Assessment of Probiotic Potential of Saccharomyces Cerevisiae DABRP5 Isolated from Bollo Batter, a Traditional Goan Fermented Food.

Bollo is a traditional Goan fermented food in which coarse wheat/wheat and finger millet is leavened with toddy. We here isolated 42 yeast strains from Bollo batter. Initial screening of the isolates with probiotic properties yielded four yeast isolates (DABRP1, DABRP2, DABRP5 and DABRP12). These isolates exhibited tolerance to high bile salt concentration and acidic pH and resistance to various antibiotics, which indicated their probiotic nature. All these yeast isolates were identified as Saccharomyces cerevisiae through D1D2-LSU-rDNA sequencing. These yeast isolates also showed higher percent hydrophobicity towards chloroform followed by n-hexadecane and o-xylene indicating their mucosal surface-adhesive property. To evaluate the safety of the isolates for them to be called as generally recognized as safe, the pathogenic behavior of the isolates determined through the in vitro hemolysis assay and evaluation of DNase and gelatinase activities. None of the isolates exhibited hemolysis or produced DNase or gelatinase and thus were considered potentially safe. In terms of beneficial effects, the most potent isolate S. cerevisiae DABRP5 showed antibacterial activity against the test pathogens. It also showed excellent antioxidant activity with DPPH free radical scavenging activity of 68.85 ± 0.69%, anti-inflammatory activity with 60.39 ± 0.34% inhibition of protein denaturation, and antidiabetic activity with 71.75 ± 0.45% inhibition of α-amylase activity. The isolate produced α-amylase, lipase, and ß-galactosidase. The probiotic potential of the isolate S. cerevisiae DABRP5 was similar to that of the reference strain (Saccharomyces boulardii CNCM I-745) used in this study. The results thus indicate that yeast isolates from Bollo batter have probiotic potential.

Suhomyces drosophilae sp. nov., isolated from Drosophila flies feeding on a stinkhorn mushroom.

The majority of Suhomyces species have been isolated from fungus-feeding insects and particularly from the gut of beetles. In the present study, seven yeast strains were isolated from the gut of Drosophila species feeding on gleba, the spore-bearing inner mass, of a stinkhorn mushroom belonging to the family Phallaceae. Based on phenotypic, biochemical characterization and sequence analysis of the D1/D2 region of the large subunit rRNA gene and the internal transcribed spacer (ITS) region, two of these yeast strains, DGY3 and DGY4, represented a novel species of the genus Suhomyces. The novel species is closely related to an undescribed species of Candida ST-370 (DQ404513) and with Suhomyces canberraensis, wherein, the novel species differs from S. canberraensis by 40 nucleotide substitutions and three gaps (7.7 % sequence variation) in the D1/D2 region and 50 nucleotide substitutions and seven gaps (13.7 % sequence variation) in the ITS region. Several morphological and physiological differences were also observed between S. canberraensis and the strains obtained during this study. These data support the proposal of Suhomyces drosophilae as a novel species, with DGY3T as the holotype and CBS 16329T and MCC 1871T as ex-type strains.