The fate of acetic acid during glucose co-metabolism by the spoilage yeast Zygosaccharomyces bailii

Zygosaccharomyces bailii is one of the most widely represented spoilage yeast species, being able to metabolise acetic acid in the presence of glucose. To clarify whether simultaneous utilisation of the two substrates affects growth efficiency, we examined growth in single- and mixed-substrate cultures with glucose and acetic acid. Our findings indicate that the biomass yield in the first phase of growth is the result of the weighted sum of the respective biomass yields on single-substrate medium, supporting the conclusion that biomass yield on each substrate is not affected by the presence of the other at pH 3.0 and 5.0, at least for the substrate concentrations examined. In vivo(13)C-NMR spectroscopy studies showed that the gluconeogenic pathway is not operational and that [2-(13)C]acetate is metabolised via the Krebs cycle leading to the production of glutamate labelled on C(2), C(3) and C(4). The incorporation of [U-(14)C]acetate in the cellular constituents resulted mainly in the labelling of the protein and lipid pools 51.5% and 31.5%, respectively. Overall, our data establish that glucose is metabolised primarily through the glycolytic pathway, and acetic acid is used as an additional source of acetyl-CoA both for lipid synthesis and the Krebs cycle. This study provides useful clues for the design of new strategies aimed at overcoming yeast spoilage in acidic, sugar-containing food environments. Moreover, the elucidation of the molecular basis underlying the resistance phenotype of Z. bailii to acetic acid will have a potential impact on the improvement of the performance of S. cerevisiae industrial strains often exposed to acetic acid stress conditions, such as in wine and bioethanol production.This work was supported by Fundacao para a Ciencia e Tecnologia (FCT), Portugal Grant PTDC/AGR-ALI/102608/2008 and by project FCOMP-01-0124-FEDER- 007047 and by FEDER through POFC - COMPETE and national funds from FCT - project PEst-C/BIA/UI4050/2011. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Reducing Bacterial Contamination in Fuel Ethanol Fermentations by Ozone Treatment of Uncooked Corn Mash

Ozonation of uncooked corn mash from the POET BPX process was investigated as a potential disinfection method for reducing bacterial contamination prior to ethanol fermentation. Corn mash (200 g) was prepared from POET ground corn and POET corn slurry and was ozonated in 250 mL polypropylene bottles. Lactic and acetic acid levels were monitored daily during the fermentation of ozonated, aerated, and nontreated corn mash samples to evaluate bacterial activity. Glycerol and ethanol contents of fermentation samples were checked daily to assess yeast activity. No yeast supplementation, no addition of other antimicrobial agents (such as antibiotics), and spiking with a common lactic acid bacterium found in corn ethanol plants,Lactobacillus plantarum, amplified the treatment effects. The laboratory-scale ozone dosages ranged from 26–188 mg/L, with very low estimated costs of $0.0008–0.006/gal ($0.21–1.6/m3) of ethanol. Ozonation was found to decrease the initial pH of ground corn mash samples, which could reduce the sulfuric acid required to adjust the pH prior to ethanol fermentation. Lactic and acetic acid levels tended to be lower for samples subjected to increasing ozone dosages, indicating less bacterial activity. The lower ozone dosages in the range applied achieved higher ethanol yields. Preliminary experiments on ozonating POET corn slurry at low ozone dosages were not as effective as using POET ground corn, possibly because corn slurry samples contained recycled antimicrobials from the backset. The data suggest additional dissolved and suspended organic materials from the backset consumed the ozone or shielded the bacteria.Reprinted with permission from J. Agric. Food Chem., 2015, 63 (21), pp 5239–5248. doi: 10.1021/acs.jafc.5b00563. Copyright 2015 American Chemical Society.</p

Assessment of a microalgae pond for post-treatment of the effluent from an anaerobic fixed bed reactor treating distillery wastewater

An evaluation of the performance of a laboratory-scale microalgae pond treating effluent from an anaerobic fixed bed reactor digesting distillery wastewater was carried out. The microalgae pond operated with an effluent recycling (R) of 10:1 with respect to the influent and at surface organic loading rates of 418 kg COD ha-1 day-1 and 92 kg BOD5 ha-1 day-1. During the experiment total chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total solids (TS), total suspended solids (TSS), volatile suspended solids (VSS), total nitrogen, ammonia, total phosphorus, orthophosphate, total chlorophyll (CT) and chlorophyll a (Ca) concentrations were monitored. Overall COD and BOD5 removal efficiencies of 98.2 % and 98.8 % were obtained. The global solids removal efficiencies were of 93.2%, 92.6 % and 97.6 % for TS, TSS and VSS, respectively. The removal efficiencies obtained for organic nitrogen and ammonia were 90.2% and 84.1%, respectively. Finally, the removals for total phosphorus and orthophosphate were 85.5% and 87.3%, respectively. It was demonstrated that microalgae grew in this waste by determination of the total chlorophyll and chlorophyll a in the effluent