接合によりグルコアミラーゼ遺伝子STA1が発現したビール酵母の育種

Standard brewing yeast cannot utilize larger oligomers or dextrins, which represent about 25% of wort sugars. A brewing yeast strain that could ferment these additional sugars to ethanol would be useful for producing low-carbohydrate diabetic or low-calorie beers. In this study, a brewing yeast strain that secretes glucoamylase was constructed by mating. The resulting Saccharomyces cerevisiae 278/113371 yeast was MATa/ diploid, but expressed the glucoamylase gene STA1. At the early phase of the fermentation test in malt extract medium, the fermentation rate of the diploid STA1 strain was slower than those of both the parent strain S. cerevisiae MAFF113371 and the reference strain bottom-fermenting yeast Weihenstephan 34/70. At the later phase of the fermentation test, however, the fermentation rate of the STA1 yeast strain was faster than those of the other strains. The concentration of ethanol in the culture supernatant of the STA1 yeast strain after the fermentation test was higher than those of the others. The concentration of all maltooligosaccharides in the culture supernatant of the STA1 yeast strain after the fermentation test was lower than those of the parent and reference strains, whereas the concentrations of flavor compounds in the culture supernatant were higher. These effects are due to the glucoamylase secreted by the constructed STA1 yeast strain. In summary, a glucoamylase-secreting diploid yeast has been constructed by mating that will be useful for producing novel types of beer owing to its different fermentation pattern and concentrations of ethanol and flavor compound

Feed intake pattern, behaviour, rumen characteristics and blood metabolites of finishing beef steers offered total mixed rations constituted at feeding or ensiling

peer-reviewedTwo experiments were undertaken. In Experiment 1, behaviour, intake pattern and blood metabolites, were recorded for steers offered total mixed rations (TMR) based on grass silage and concentrates, and constituted either at ensiling (E-TMR) or feedout (F-TMR). Fourteen continental crossbred steers (mean starting weight 505 (s.d. 41.5) kg) were assigned to each of the following eight treatments: grass silage offered ad libitum (SO), E-TMR diets constituted in approximate dry matter (DM) ratios of grass:concentrates of 75:25 (EL), 50:50 (EM) and 25:75 (EH), F-TMR diets constituted in approximate DM ratios of grass silage:concentrates of 75:25 (FL), 50:50 (FM) and 25:75 (FH), and finally concentrates ad libitum (AL). Total DM intake increased linearly (P < 0.001) and the time spent eating and ruminating decreased linearly (P < 0.001) with increasing concentrate proportion. Animals on the F-TMR diets had higher total DM intakes (P < 0.05) and plasma glucose (P < 0.05) and urea (P < 0.001) concentrations than animals on the corresponding E-TMR diets. No effect of method of feed preparation on intake pattern or behaviour was recorded. In Experiment 2, four ruminally cannulated Holstein-Friesian steers of mean initial live weight 630 (s.d. 23.2) kg were used to evaluate rumen characteristics for four of the above diets (FL, EL, FH and EH) in a 4 × 4 latin square design. Higher concentrate diets resulted in lower rumen pH (P < 0.05), higher lactic acid (P < 0.001) and ammonia (P < 0.05) concentrations and lower acetate:propionate (P < 0.05). F-TMR was associated with a higher (P < 0.05) rumen volatile fatty acid concentration but no difference in other rumen fermentation characteristics compared to E-TMR. Concentrate proportion and method of feed preparation had no effect (P > 0.05) on rumen pool sizes but animals consuming the high concentrate diet had a faster (P < 0.05) rumen passage rate of NDF than animals on the low concentrate diet.B. Cummins was in receipt of a Teagasc Walsh Fellowship

Milk production from leguminous forage, roots and potatoes

The aim of the present work was to investigate the effects of replacing grain concentrates with roots and potatoes in dairy cow diets based upon large amounts of grass/alfalfa silage. The emphasis was on the possible improvement of microbial protein synthesis and nitrogen balance. Alfalfa dominated silage has a large excess of ruminally degradable protein that must be balanced with feed carbohydrates to avoid urinary nitrogen losses. The effects on ruminal fermentation pattern, intake and production were also studied. The thesis is based on two batch culture in vitro experiments and three animal experiments. The in vitro experiments compared fodder beets, barley/oats and raw, boiled or frozen potatoes as supplements to a silage diet incubated with rumen fluid from cows fed different diets. With respect to amounts fermented during 5 h incubation, supplements were ranked (P barley/oats > raw potatoes = frozen potatoes = unsupplemented silage. Substrates were numerically ranked in the same order with respect to microbial protein production, but due to larger variation they could only be divided into two groups, where fodder beets, boiled potatoes and barley/oats gave microbial yields not different from each other, but higher than for raw potatoes, frozen potatoes or unsupplemented silage. Butyrate proportion was little affected by incubation substrate but fodder beets fed to rumen fluid donor cows increased butyrate molar proportion in vitro from 10.7 to 13.0%. A change-over design experiment compared barley supplementation with fodder beet and potato supplementation of a silage diet for lactating cows. The fodder beet/potato diet lowered ad libitum silage intake by 0.9 kg DM/d and milk yield decreased correspondingly by 1.7 to 2.3 kg/d. Microbial protein production and nitrogen balance were not increased by the fodder beet supplementation, but a part of N excretion was redirected from urine to feces. Fodder beets tended to decrease the ratio lipogenic/glucogenic VFA, by increasing propionate and butyrate at the expense of acetate. In an intake experiment, most of the cows consumed the maximum allowance of fodder beets (4.6 kg DM/d) while there was a huge variation in the potato intake. A more synchronous feeding of degradable protein and readily available carbohydrates lowered the urinary nitrogen loss and increased allantoin excretion numerically but not significantly. A close correlation (R2 = 0.94) was found between total urinary N excretion and the ratio urea/creatinine in urine, which implies that spot sampling of urine may be a way to facilitate N balance measurements in lactating cows. In conclusion, a full replacement of grain by roots and potatoes can be done and the effects will be lowered urinary N losses but also a reduction in silage consumption and hence also milk production

Isolation and characterisation of Sri Lankan yeast germplasm and its evaluation for alcohol production

Use of inferior yeast cultures represents one of the reasons for low fermentation efficiencies in Sri Lankan alcohol distilleries that use sugarcane molasses. The present study isolated and characterised yeast strains found in natural environments in Sri Lanka and evaluated their performance under laboratory conditions in an effort to select superior strains for industrial fermentations. Yeasts were characterised based on morphological and physiological features such as sugar fermentation and nitrate assimilation. Ethanol production, alcohol tolerance and growth rate of the most promising strains were monitored following laboratory fermentations of molasses. Over a thousand yeast cultures were collected and screened for fermentative activity and a total of 83 yeast isolates were characterised as higher ethanol producers. Most of these belonged to the genus Saccharomyces. Certain strains produced over 10% (v/v) alcohol in molasses media during 72 h laboratory fermentations. Only two strains, SL-SRI-C-102 and 111, showed an appreciable fermentation efficiency of about 90%. The latter strain produced the highest level of ethanol, 11% (v/v) within a 48 h fermentation and exhibited improved alcohol tolerance when compared with the baker's yeast strains currently used in Sri Lankan alcohol distilleries. This study highlights the benefits of exploiting indigenous yeasts for industrial fermentation processes

Electromagnetic Actuated stirring in Microbioreactors enabling easier multiplexing & flexible device design

This paper was presented at the 4th Micro and Nano Flows Conference (MNF2014), which was held at University College, London, UK. The conference was organised by Brunel University and supported by the Italian Union of Thermofluiddynamics, IPEM, the Process Intensification Network, the Institution of Mechanical Engineers, the Heat Transfer Society, HEXAG - the Heat Exchange Action Group, and the Energy Institute, ASME Press, LCN London Centre for Nanotechnology, UCL University College London, UCL Engineering, the International NanoScience Community, www.nanopaprika.eu.The development of a novel electromagnetically (EM) actuated stirring method, for use in microbioreactors, is reported. Mixing in microbioreactors is critical to ensure even distribution of nutrients to microorganisms and cells. Magnetically driven stirrer bars or peristaltic mixing are the most commonly utilised mixing methods employed in completely liquid-filled microbioreactors. However the circular reactor shape required for mixing with a stirrer bar and frequently used for peristaltically mixed microbioreactors presents difficulties for bubble-free priming in a microfluidic bioreactor. Moreover the circular shape and the hardware required for both types of mixing reduces the potential packing density of multiplexed reactors. We present a new method of mixing, displaying design flexibility by demonstrating mixing in circular and diamond-shaped reactors and a duplex diamond reactor and fermentation of the gram-positive bacteria S. carnosus in a diamond-shaped microbioreactor system. The results of the optimisation of this mixing method for performing fermentations alongside both batch and continuous culture fermentations are presented

Effects of long-term soluble vs. insoluble dietary fiber intake on high-fat diet-induced obesity in C57BL/6J mice

Although most of the proposed beneficial effects of fiber consumption have been attributed to viscous and gel-forming properties of soluble fiber, it is mainly insoluble cereal fiber and whole grains that are strongly associated with reduced diabetes risk in prospective cohort studies, indicating that other unknown mechanisms are likely to be involved. We performed a long-term study investigating potential protective effects of adding soluble guar fiber (10% w/w) vs. insoluble cereal fiber (10% w/w) to an isoenergetic and macronutrient matched high-fat diet in obesity-prone C57BL/6J mice. After 45 weeks, mice fed soluble vs. insoluble fiber showed both significantly increased body weight (41.8±3.0 vs. 33.6±1.5 g, P=.03) and elevated markers of insulin resistance. In mice fed soluble fiber, energy loss via the feces was significantly lower and colonic fermentation with production of short chain fatty acids (SCFA) was markedly increased. Gene expression analysis in white adipose tissue showed significantly increased levels of the fatty acid target G-protein coupled receptor-40 in soluble fiber-fed mice. Liver gene expression in the insoluble fiber group showed a pattern consistent with increased fatty acid oxidation. The present results show that soluble vs insoluble dietary fiber added to a high-fat, Western-style diet differently affected body weight and estimates of insulin sensitivity in obesity-prone mice. Soluble fiber intake with increased SCFA production significantly contributed to digested energy, thereby potentially outweighing the well known short-term beneficial effects of soluble fiber consumption

Mathematical Model of \u3cem\u3eChlorella minutissima\u3c/em\u3e UTEX2341 Growth and Lipid Production Under Photoheterotrophic Fermentation Conditions

To reduce the cost of algal biomass production, mathematical model was developed for the first time to describe microalgae growth, lipid production and glycerin consumption under photoheterotrophic conditions based on logistic, Luedeking–Piret and Luedeking–Piret-like equations. All experiments were conducted in a 2 L batch reactor without considering CO2 effect on algae’s growth and lipid production. Biomass and lipid production increased with glycerin as carbon source and were well described by the logistic and Luedeking–Piret equations respectively. Model predictions were in satisfactory agreement with measured data and the mode of lipid production was growth-associated. Sensitivity analysis was applied to examine the effects of certain important parameters on model performance. Results showed that S0, the initial concentration of glycerin, was the most significant factor for algae growth and lipid production. This model is applicable for prediction of other single cell algal species but model testing is recommended before scaling up the fermentation of process

Cereal non-starch polysaccharides in pig diets

This thesis is based on three different studies comprising the weaning and the growing period, aiming at monitoring the influence of cereal non-starch polysaccharides (NSP) and dietary enzyme supplementation on gastro-intestinal processes in pigs. The diets were based on cereals and cereal by-products, and composed to contain different amounts of total as well as soluble NSP. Results from these studies have shown that with increased NSP content, the reduction in digestibility of organic matter (OM) was twice as high in the small intestine than in the total tract, both in newly weaned piglets and growing pigs. An increased proportion of insoluble NSP decreased the digestibility of OM and fibre components in the small intestine of the newly weaned piglets and in the total tract of growing pigs. The gut environment, as described by content and proportions of organic acids (OA) and pH, as well as total microbial populations and coliform diversity, was altered by NSP content and solubility, whereas enzyme supplementation influenced the distribution of OA in the small intestine. PVTC-cannulation did not influence the coliform flora, and results obtained from PVTC-cannulated pigs were concluded to reflect true intestinal conditions. In conclusion, these results indicate that the dietary content of total and soluble NSP influence gastro-intestinal processes such as digestion site, gut environment and microbial populations in different ways in newly weaned piglets and in growing pigs. Therefore, NSP constitute an important tool with possibilities to influence gut health in pigs, and may therefore offer prospects to optimise the feed for pigs of different age

The role of nitrogen uptake on the competition ability of three vineyard Saccharomyces cerevisiae strains

Three vineyard strains of Saccharomyces cerevisiae, P301.4, P304.4 and P254.12, were assayed in comparison with a commercial industrial strain, QA23. The aim was to understand if nitrogen availability could influence strain competition ability during must fermentation. Pairwise-strain fermentations and co-fermentations with the simultaneous presence of the four strains were performed in synthetic musts at two nitrogen levels: control nitrogen condition (CNC) that assured the suitable assimilable nitrogen amount required by the yeast strains to complete the fermentation and low nitrogen condition (LNC) where nitrogen is present at very low level. Results suggested a strong involvement of nitrogen availability, as the frequency in must of the vineyard strains, respect to QA23, in LNC was always higher than that found in CNC. Moreover, in CNC only strain P304.4 reached the same strain frequency as QA23. P304.4 competition ability increased during the fermentation, indicating better performance when nitrogen availability was dropping down. P301.4 was the only strain sensitive to QA23 killer toxin. In CNC, when it was co-inoculated with the industrial strain QA23, P301.4 was never detected. In LNC, P301.4 after 12 h accounted for 10% of the total population. This percentage increased after 48 h (20%). Single-strain fermentations were also run in both conditions and the nitrogen metabolism further analyzed. Fermentation kinetics, ammonium and amino-acid consumptions and the expression of genes under nitrogen catabolite repression evidenced that vineyard yeasts, and particularly strain P304.4, had higher nitrogen assimilation rate than the commercial control. In conclusion, the high nitrogen assimilation rate seems to be an additional strategy that allowed vineyard yeasts successful competition during the growth in grape musts