andSaccharomyces Cerevisiae Research Articles

Chemical Composition and Biological Activities of Essential Oil of Elsholtzia ciliata obtained at Central Highlands of Vietnam’s Two Seasons

From the essential oil of the whole plant from Elsholtzia ciliata, obtained by hydrodistillation and then using gas chromatography/mass spectrometry to analyze and identify 42 constituents, carveol (26.89%) and z-citral (29.35%) main contents (collected in the dry season time); and 46 constituents, cis-ocimene (17.95%), carveol (14.84%), Z-citral (17.01%), isocaryophyllene (13.19%) and trans-caryophyllene (9.76%) as main contents (collected in the rainy season time). The major essential oils in dry season exhibited good inhibition activity on the Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Candida albicans with MIC value of 200 μg/mL; exhibited weak cytotoxic activity on the PC-3 line (IC50 66.34 μg/mL) and A549 line (IC50 93.11 μg/mL). The essential oils in rainy season exhibited good inhibition activity on the Escherichia coli, Bacillus subtillis and Saccharomyces cerevisiae with MIC value of 200 μg/mL and Candida albicans with MIC value of 100 μg/mL; displayed good cytotoxic activity on the PC-3 line (IC50 30.62 μg/mL) and Hep-G2 line (IC50 22.78 μg/mL), however have weak cytotoxic activity on the A549 line (IC50 85.35 μg/mL).

옻나무 추출물의 효모-젖산균 혼합발효를 통한 고농도 γ-aminobutyric acid 생산

Toxicodendron vernicifluum extract (TVE) was co-fermented by Saccharomyces cerevisiae and Lactobacillus plantarum EJ2014 to produce higher content of gamma-aminobutyric acid (GABA). The TVE was cultured at 30°C for 2 days under static conditions with sucrose 5%, yeast extract 1%, and Saccharomyces cerevisiae starter 1%. The second lactic acid fermentation after the primary first yeast fermentation was performed at 30°C for 7 days after adding monosodium L-glutamate (MSG) 2.5%, lactose 2.5% and lactic acid bacteria (LAB) starter 1%. The first fermented TVE showed pH 4.6, 0.1% acidity and viable cell counts of S. cerevisiae 7.63 log CFU/mL. After the second fermentation, co-fermented TVE showed pH 4.64, 0.68% acidity, and viable cell count of L. plantarum EJ2014 7.68 log CFU/mL. During the co-fermentation with S. cerevisiae and L. plantarum EJ2014, the MSG was completely converted, yielding 2.5% GABA. Conclusively, co-fermentation of TVE was successfully performed with S. cerevisiae and L. plantarum EJ2014, resulting in a higher production of GABA and probiotics. The final fermented TVE could be utilized as a multi-functional ingredient in food and bio-industries.

창포식초 생산을 위한 누룩, 알코올 및 초산발효 조건 최적화

In this study, we investigated the quality characteristics of medicinal vinegar prepared from the roots of Acorus gramineus. We used barley alcohol beverage along with both Rhizopus oryzae 82-7 and Saccharomyces cerevisiae YM33 and vinegar culture with Acetobacter pasteurianus A11-2 under the optimal conditions for acetic acid fermentation. We manufactured the barley alcohol beverage with 11% alcohol content for fermentation with ‘Nuruk’ and Saccharomyces cerevisiae YM33 at 25°C. When fermented vinegar from Acorus gramineus roots for 10 days at 30°C with a starter, the pH and total acidity were 3.48 and 4.44%, respectively, and the alcohol in vinegar was totally consumed. Besides, the vinegar showed total polyphenol content of 870.13 mg TAE/mL at the 15th day, along with significant antioxidant activity. GC-MS analysis revealed that the major volatiles in vinegar were methyl salicylate and 2-ethylhexanol, which have odors like those of mint and rose, respectively. In contrast, the ethanol, 3-methyl-1-butanol, and 2-methyl-1-propanol contents, which were high in the barley alcohol beverage, decreased in the vinegar that was fermented by Acorus gramineus. Consequently, the vinegar fermented by Acorus gramineus showed unique aromas and high antioxidant activity.

Incredible edible selenium nanoparticles produced by food-grade microorganisms

Background: Microorganisms commonly employed in food industry, such as Lactobacillus plantarum and Saccharomyces cerevisiae, are also excellent natural nanotechnologists. They reduce selenite (SeO3 2- ) to form nanoparticles of red selenium (Se0 ) of exceptional quality and with interesting physical and (bio-)chemical properties. Objectives: The production of these nanoparticles has been studied in several relevant microorganisms to gain a better picture of the overall properties and quality of these particles, possible differences between producers, ease of production and, in particular, biological activity. Methods: Several common microorganisms, namely L. plantarum, S. cerevisiae and Escherichia coli have been cultured under standard conditions and 1mM concentrations of SeO3 2- have been converted to red particles of elemental selenium. These particles have been characterized extensively with respect to uniformity, size, shape, consistency and, in particular, biological activity against infectious microbes. Results: Highly uniform amorphous spherical particles of 100 nm to 200 nm in diameter could be produced by several microorganisms, including Lactobacillus. Although originating in bacteria and yeast, these particles exhibit antimicrobial activity when employed at concentrations of around 100 µM. This activity may in part be due to the inherent chemistry of selenium and /or of the protein coating of the particles. Interestingly, yeast also forms larger rod-like structures. These micro-needles with around 85 nm in diameter and up to 3 µm in length exhibit considerable antibacterial activity, possibly resulting from additional, physical interactions with cellular structures. Conclusion: Common microorganisms traditionally employed in the preparation of food produce nanoparticles of selenium which may be harvested and explored as natural antimicrobial agents or antioxidants. These particles provide a fine example of and lead for natural nanotechnology with biological activity and applications in food and food supplementation, medicine, agriculture and cosmetics.

Celiac disease-specific and inflammatory bowel disease-related antibodies in patients with recurrent aphthous stomatitis

The etiology of recurrent aphthous stomatitis (RAS) remains unknown. RAS can be presented as primary, idiopathic condition and as a secondary RAS, which is associated with a systemic disease. The aim of our study was to evaluate the presence and concentrations of antibodies specific for celiac disease (CeD) and antibodies related to inflammatory bowel diseases (IBD) in patients with RAS without gastrointestinal symptoms. Antibodies against tissue transglutaminase (anti-tTG), deaminated gliadin peptides (DGP), deaminated gliadin-analogous fragments (anti-GAF-3X) and Saccharomyces cerevisiae (ASCA) were determined by ELISA and antineutrophil cytoplasmic antibodies (ANCA) by indirect immunoflurescence (IIF) in 57 patients with RAS and 60 control subjects. The prevalence of CeD specific antibodies did not differ between RAS patients and controls. However, the concentrations of IgA anti-tTG, IgA anti-GAF-3X antibodies in patients with RAS were significantly higher compared to controls (p = 0.002 and p = 0.04 respectively). Histological changes consistent with CeD were confirmed by duodenal biopsy in one RAS patient with highly positive IgA anti-tTG, anti-GAF-3X and anti-DGP antibodies. Higher prevalence along with higher concentrations of IgG ASCA were found in RAS patients compared to controls (p < 0.01). Patients with positive IgG ASCA in the absence of clinical symptoms decided not to pursue any further testing. Dysfunction of oral mucosa and the exposure to various antigens might be a reason for the loss of tolerance resulting in increased production of autoantibodies. It seems likely that antibodies are markers of aberrant immune response, rather than key effectors involved in the pathogenesis of the disease.

Pulmonary Deposition and Elimination of Liposomal Amikacin for Inhalation and Effect on Macrophage Function after Administration in Rats.

Pulmonary nontuberculous mycobacterial (PNTM) infections represent a treatment challenge. Liposomal amikacin for inhalation (LAI) is a novel formulation currently in development for the treatment of PNTM infections. The pulmonary deposition and elimination of LAI and its effect on macrophage function were evaluated in a series of preclinical studies in healthy rats. The pulmonary deposition of LAI was evaluated in female rats (n = 76) treated with LAI by nebulizer at 10 mg/kg of body weight per day or 90 mg/kg per day for 27 days, followed by dosing of dually labeled LAI (LAI with a lipid label plus an amikacin label) on day 28 with subsequent lung histological and amikacin analyses. In a separate study for assessment of alveolar macrophage function, rats (n = 180) received daily treatment with LAI at 90 mg/kg per day or 1.5% saline over three 30-day treatment periods followed by 30-day recovery periods; phagocytic and Saccharomyces cerevisiae (yeast) killing capabilities and inflammatory mediator release were assessed at the end of each period. LAI demonstrated equal dose-dependent deposition across all lung lobes and regions. Lipid and amikacin labels showed diffuse extracellular colocalization, followed by macrophage uptake and gradual amikacin elimination. Macrophages demonstrated accumulation of amikacin during treatment periods and nearly complete elimination during recovery periods. No evidence of an inflammatory response was seen. No differences in microsphere uptake or yeast killing were seen between LAI-treated and control macrophages. Neither LAI-treated nor control macrophages demonstrated constitutive inflammatory mediator release; however, both showed normal mediator release on lipopolysaccharide stimulation. LAI is readily taken up by macrophages in healthy rats without compromising macrophage function.

Production of Palmitoleic and Linoleic Acid in Oleaginous and Nonoleaginous Yeast Biomass

We investigated the possibility of utilizing both oleaginous yeast species accumulating large amounts of lipids (Yarrowia lipolytica, Rhodotorula glutinis, Trichosporon cutaneum, and Candida sp.) and traditional biotechnological nonoleaginous ones (Kluyveromyces polysporus, Torulaspora delbrueckii, and Saccharomyces cerevisiae) as potential producers of dietetically important major fatty acids. The main objective was to examine the cultivation conditions that would induce a high ratio of dietary fatty acids and biomass. Though genus-dependent, the type of nitrogen source had a higher influence on biomass yield than the C/N ratio. The nitrogen source leading to the highest lipid accumulation was potassium nitrate, followed by ammonium sulfate, which is an ideal nitrogen source supporting, in both oleaginous and nonoleaginous species, sufficient biomass growth with concomitantly increased lipid accumulation. All yeast strains displayed high (70–90%) content of unsaturated fatty acids in total cell lipids. The content of dietary fatty acids of interest, namely, palmitoleic acid and linoleic acid, reached in Kluyveromyces and Trichosporon strains over 50% of total fatty acids and the highest yield, over 280 mg per g of dry cell weight of these fatty acids, was observed in Trichosporon with ammonium sulfate as nitrogen source at C/N ratio 70.

Quorum Sensing-Mediated, Cell Density-Dependent Regulation of Growth and Virulence in Cryptococcus neoformans

ABSTRACTQuorum sensing (QS) is a cell density-dependent mechanism of communication between microorganisms, characterized by the release of signaling molecules that affect microbial metabolism and gene expression in a synchronized way. In this study, we investigated cell density-dependent behaviors mediated by conditioned medium (CM) in the pathogenic encapsulated fungus Cryptococcus neoformans. CM produced dose-dependent increases in the growth of planktonic and biofilm cells, glucuronoxylomannan release, and melanin synthesis, important virulence attributes of this organism. Mass spectrometry revealed the presence of pantothenic acid (PA) in our samples, and commercial PA was able to increase growth and melanization, although not to the same extent as CM. Additionally, we found four mutants that were either unable to produce active CM or failed to respond with increased growth in the presence of wild-type CM, providing genetic evidence for the existence of intercellular communication in C. neoformans. C. neoformans CM also increased the growth of Cryptococcus albidus, Candida albicans, and Saccharomyces cerevisiae. Conversely, CM from Cryptococcus albidus, C. albicans, S. cerevisiae, and Sporothrix schenckii increased C. neoformans growth. In summary, we report the existence of a new QS system regulating the growth and virulence factor expression of C. neoformans in vitro and, possibly, also able to regulate growth in other fungi.

Microbiological and physico-chemical changes during fermentation of maize for masa production

Samples of traditionally fermenting maize meal popularly called masa were collected from local producers of masa in Ibadan, Oyo State, Nigeria and were analysed for pH, total titratable acidity (TTA), microbiological changes and proximate analysis. The sensory properties (colour, texture, taste, sourness, appearance and general acceptability) were also carried out on a nine point hedonic scale. The values obtained were compared with the starter culture fermenting maize meal prepared in the laboratory. The physico-chemical changes during the traditional and the laboratory prepared fermenting maize meal were observed at 6 h interval during the fermentation process. There was decrease in pH; the pH of the traditional fermenting maize meal ranged between 5.98 ± 0.27 to 3.95 ± 0.05 while the pH of the laboratory prepared fermenting maize meal ranged between 6.4 ± 0.36 to 4.3 ± 0.03. There was increase in TTA during the fermentation period; in the traditional fermenting maize meal the TTA ranged between 0.144 ± 0.02 to 0.792 ± 0.08 and it ranged between 0.054 ± 0.02 to 0.252 ± 0.00 in the laboratory fermenting maize meal. Microorganisms associated with the traditional fermentation of masa wereLactobacillus plantarum, Pediococcus acidilactici, Lactobacillus fermentum andSaccharomyces cerevisiae. In all the fermenting maize meal samples collected and the laboratory prepared maize meal samples inoculated with starter organisms, an increase in the microbial load counts (log10cfu/ml) was observed as the fermentation time progresses, 3.02 to 6.41 (on De Mann Rogosa-Sharpe agar), 2.99 to 3.62 (on malt extract agar), 4.18 to 7.00 (on nutrient agar) during the traditional fermentation, in the laboratory prepared maize meal samples inoculated with starter organisms, the microbial load counts (log10cfu/ml) ranged between 4.01 to 9.05. The laboratory prepared maize meal samples inoculated with starter organisms only showed minimal changes in their proximate composition as compared with the traditionally fermented samples collected. The statistical analysis of the sensory attributes revealed consumer’s acceptance of the product inoculated with the starter organisms. Key words: Proximate analysis, TTA, physico-chemical changes, masa, starter organisms.

Antibody response against gastrointestinal antigens in demyelinating diseases of the central nervous system

Antibodies against gastrointestinal antigens may indicate altered microbiota and immune responses in the gut. Recent experimental data suggest a connection between gastrointestinal immune responses and CNS autoimmunity. Antibodies against gliadin, tissue transglutaminase (tTG), intrinsic factor (IF), parietal cells (PC) and Saccharomyces cerevisiae (ASCA) were screened in the sera of 45 patients with AQP4-seropositive neuromyelitis optica (NMO) and NMO spectrum diseases (NMO/NMO-SD), 17 patients with AQP4-seronegative NMO, 85 patients with clinically definite multiple sclerosis (MS), and 48 healthy controls (HC). Thirty-seven percentages of patients with AQP4-seropositive NMO/NMO-SD and 28% of patients with MS had at least one particular antibody in contrast to 8% of HC (P<0.01, respectively). Antibodies were most common (46%) in AQP4-seropositive myelitis (P=0.01 versus HS, P=0.05 versus MS). Anti-gliadin and ASCA were more frequent in the AQP4-seropositive NMO-spectrum compared to controls (P=0.01 and P<0.05, respectively). Antibody responses against gastrointestinal antigens are common in MS and AQP4-seropositive NMO/NMO-SD, especially in longitudinally extensive myelitis.

The power of yeast to model diseases of the powerhouse of the cell.

Mitochondria participate in a variety of cellular functions. As such, mitochondrial diseases exhibit numerous clinical phenotypes. Because mitochondrial functions are highly conserved between humans andSaccharomyces cerevisiae, yeast are an excellent model to study mitochondrial disease, providing insight into both physiological and pathophysiological processes.

English

Twenty six (26) strains of&nbsp;Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei,&nbsp;Enterococcus faecium, Bacillus subtilis, Bacillus coagulans,&nbsp;andSaccharomyces cerevisiae&nbsp;from Korean Collection for Type Cultures (KCTC) were evaluated in this experiment for the development of green tea based probiotics production and their inhibition effect against pathogenic bacteria was examined. The strains were first tested for acid and bile tolerance, and the higher tolerance strains were subsequently analyzed for heat tolerance and susceptibility to green tea extracts. Among the tested strains,&nbsp;L. acidophilus&nbsp;KCTC 3111,&nbsp;L. plantarum&nbsp;KCTC 3104,&nbsp;B. subtilis&nbsp;KCTC 3239,&nbsp;B. coagulans&nbsp;KCTC 1015, and&nbsp;S. cerevisiae&nbsp;KCTC 7915 which displayed a high tolerance to acid, bile, and heat, and no susceptibility to green tea extract, were selected to produce fermented green tea probiotics. Associative effects of green tea probiotics andEscherichia coli&nbsp;S93 F5, S99 LT, S170 F41, and S171 F6 revealed that&nbsp;E. coli&nbsp;numbers had gradually decreased compared to the control. Stability results of&nbsp;Lactobacillus&nbsp;spp. and&nbsp;Bacillus&nbsp;spp. in green tea probiotics showed that they were stable at 108&nbsp;CFU/g during eight weeks of storage. Therefore, it is expected that green tea probiotics may provide a potential feed additive for livestock. &nbsp; Key words:&nbsp;Green tea, probiotics, tolerance,&nbsp;E. coli, feed additive, livestock.

Sulfurous Gases As Biological Messengers and Toxins: Comparative Genetics of Their Metabolism in Model Organisms

Gasotransmitters are biologically produced gaseous signalling molecules. As gases with potent biological activities, they are toxic as air pollutants, and the sulfurous compounds are used as fumigants. Most investigations focus on medical aspects of gasotransmitter biology rather than toxicity toward invertebrate pests of agriculture. In fact, the pathways for the metabolism of sulfur containing gases in lower organisms have not yet been described. To address this deficit, we use protein sequences from Homo sapiens to query Genbank for homologous proteins in Caenorhabditis elegans, Drosophila melanogaster, and Saccharomyces cerevisiae. In C. elegans, we find genes for all mammalian pathways for synthesis and catabolism of the three sulfur containing gasotransmitters, H2S, SO2 and COS. The genes for H2S synthesis have actually increased in number in C. elegans. Interestingly, D. melanogaster and Arthropoda in general, lack a gene for 3-mercaptopyruvate sulfurtransferase, an enzym for H2S synthesis under reducing conditions.

Yeasts colonizing the leaves of fruit trees

Yeasts were isolated from leaf surfaces of five species of fruit trees located in southwest Slovakia. One hundred and fifty five yeast strains belonging to 11 genera were isolated from 300 samples of leaves. Seventeen yeast species were identified, but only three occurred regularly:Aureobasidium pullulans, Cryptococcus laurentii, andMetschnikowia pulcherrima. Species such asHanseniaspora uvarum, Pichia anomala, Rhodotorula glutinis, andSaccharomyces cerevisiae, were isolated less frequently. We found only few differences in the yeast community isolated from leaves of different tree species although dominant species occurred regularly on the majority of leaves. Furthermore, yeast species varied throughout years. In spite of the fact that the yeast community occupying the leaves of the fruit trees was studied in the samples harvested in three localities, which are distanced from each other some kilometers, the qualitative representation of the most isolated yeasts was identical. The differences were only in the frequency of the incidence of individual species in the samples.

Determination of thein vitro activity of fluconazole against yeast strains using HPLC

The aim of this study was to evaluate thein vitro activity of fluconazole against two yeast strains by determining the fluconazole content via high-performance liquid chromatography (HPLC).Candida krusei ATCC 6258 andSaccharomyces cerevisiae were tested for fluconazole susceptibility by dilution tests. Fluconazole measurements by HPLC were performed in cultures of the strains at 6, 12, 24 and 48 h. InC. krusei ATCC 6258 cultures, 88.94% of the fluconazole was recovered at 48 h with respect to the level at time 0 h. The drug exhibited major activity at 12 and 24 h. An increase in cell number and elevated concentration of the drug were detected at 48 h. InS. cerevisiae cultures, the measurements revealed a fluconazole content of approximately 51.25% at 12, 24 and 48 h with respect to the level at time 0 h. However, the percentage of fluconazole detected and the minimal inhibitory concentration (MIC) value determined by agar dilution tests suggest a potential for fluconazole resistance in this susceptible strain. The decreases in fluconazole inC. krusei ATCC 6258 andS. cerevisiae cultures at each time point suggest a mechanism of resistance that is not correlated to efflux pumps.

Cascading failure and robustness in metabolic networks

We investigate the relationship between structure and robustness in the metabolic networks of Escherichia coli, Methanosarcina barkeri, Staphylococcus aureus, and Saccharomyces cerevisiae, using a cascading failure model based on a topological flux balance criterion. We find that, compared to appropriate null models, the metabolic networks are exceptionally robust. Furthermore, by decomposing each network into rigid clusters and branched metabolites, we demonstrate that the enhanced robustness is related to the organization of branched metabolites, as rigid cluster formations in the metabolic networks appear to be consistent with null model behavior. Finally, we show that cascading in the metabolic networks can be described as a percolation process.

Partial sequencing of the β-glucosidase-encoding gene of yeast strains isolated from musts and wines

The aim of the present work was the identification of the gene encoding for β-glucosidase and its partial sequencing in the strainsPichia anomala AL112,Hanseniaspora uvarum Y8 andSaccharomyces cerevisiae AL41. To this aim degenerated primers, designed on the basis of aminoacid similarities of four known yeast β-glucosidases, have been used in PCR amplifications. An expected fragment of about 200 bp was amplified from all the DNAs, cloned and sequenced. Sequence homology demonstrated for the first time the presence of a β-glucosidase encoding gene inHanseniaspora uvarum andSaccharomyces cerevisiae.

Evaluation of the New Micronaut- Candida System Compared to the API ID32C Method for Yeast Identification

A new system, Micronaut-Candida, was compared to API ID32C to identify 264 yeast (Candida albicans, C. parapsilosis, C. tropicalis, C. krusei, C. inconspicua, C. norvegensis, C. lusitaniae, C. guilliermondii, C. dubliniensis, C. pulcherrima, C. famata, C. rugosa, C. glabrata, C. kefyr, C. lipolytica, C. catenulata, C. neoformans, Geotrichum and Trichosporon species, Rhodotorula glutinis, and Saccharomyces cerevisiae) clinical isolates. Results were in concordance in 244 cases. Eighteen out of the 20 of discordant results were correctly identified by Micronaut-Candida but not by API ID32C, as confirmed by PCR ribotyping.

DNA Microarray Based on Arrayed-Primer Extension Technique for Identification of Pathogenic Fungi Responsible for Invasive and Superficial Mycoses

An oligonucleotide microarray based on the arrayed-primer extension (APEX) technique has been developed to simultaneously identify pathogenic fungi frequently isolated from invasive and superficial infections. Species-specific oligonucleotide probes complementary to the internal transcribed spacer 1 and 2 (ITS1 and ITS2) region were designed for 24 species belonging to 10 genera, including Candida species (Candida albicans, Candida dubliniensis, Candida famata, Candida glabrata, Candida tropicalis, Candida kefyr, Candida krusei, Candida guilliermondii, Candida lusitaniae, Candida metapsilosis, Candida orthopsilosis, Candida parapsilosis, and Candida pulcherrima), Cryptococcus neoformans, Aspergillus species (Aspergillus fumigatus and Aspergillus terreus), Trichophyton species (Trichophyton rubrum and Trichophyton tonsurans), Trichosporon cutaneum, Epidermophyton floccosum, Fusarium solani, Microsporum canis, Penicillium marneffei, and Saccharomyces cerevisiae. The microarray was tested for its specificity with a panel of reference and blinded clinical isolates. The APEX technique was proven to be highly discriminative, leading to unequivocal identification of each species, including the highly related ones C. parapsilosis, C. orthopsilosis, and C. metapsilosis. Because of the satisfactory basic performance traits obtained, such as reproducibility, specificity, and unambiguous interpretation of the results, this new system represents a reliable method of potential use in clinical laboratories for parallel one-shot detection and identification of the most common pathogenic fungi.

Development of a serial bioreactor system for direct ethanol production from starch usingAspergillus niger andSaccharomyces cerevisiae

Aspergillus niger hyphae were found to grow with unliquefied potato starch under aerobic conditions, but did not grow under anaerobic conditions. The raw culture ofA. niger catalyzed saccharification of potato starch to glucose, producing approximately 12 g glucose/L/day/ The extracellular enzyme activity was decreased in proportion to incubation time, and approximately 64% of initial activity was maintained after 3 days. At 50°C,A. niger hyphae growth stopped, while the extracellular enzyme activity peaked. On the basis of theA. niger growth property and enzyme activity, we designed a serial bioreactor system composed of four different reactors. Fungal hyphae were cultivated in reactor I at 30°C, uniquefied starch was saccharified to glycose by a fungal hyphae culture in reactors II and III at 50°C, and glucose was fermented to ethanol bySaccharomyces cerevisiae in reactor IV. The total glucose produced by fungal hyphae in reactor I and saccharification in reactor II was about 42 g/L/day. Ethanol production in reactor IV was approximately 22 g/L/day, which corresponds to about 79% of the theoretical maximum produced from 55 g starch/L/day.