Application of rotary microfiltration in debittering process of spent brewer's yeast

This study concerns the production of yeast extract from spent brewer's yeast using rotary microfiltration as a means to combine debittering and cell debris separation into a single step, without using a toxic alkali wash. The pH of yeast homogenate was found to affect protein yield and bitterness of the product. Rotary filtration of yeast homogenate at various pHs resulted in different percent protein transmissions. These were found to be 5.05%, 9.83%, and 30.83% for pH 5, 6, and 7.5, respectively. The bitterness concentration in the permeate was also found to be higher at higher pHs. Autolysis of the cell homogenate prior to filtration increased protein yield and decreased bitterness considerably. At pH 5.5, the protein transmission was increased to 60% and debittering efficiency was increased from 59% to 86%. The permeate flux and protein productivity could be further increased by increasing the rotational speed, but this resulted in a decrease in debittering efficiency. Thus, the rotational speed should be carefully selected to compromise between the yield and product quality. Furthermore, for the tested rotational speeds of 600 and 1000 rpm, the change in feed flow rate from 11 to 35 L h(-1) changes the flow behavior from turbulent vortex flow to laminar vortex flow, thus decreasing the flux and protein productivity.     

Application of a simple yeast extract from spent brewer's yeast for growth and sporulation of Bacillus thuringiensis subsp. kurstaki: a Physiological Study

The suitability of using a simple brewer's yeast extract (BYE), prepared by autolysis of complete beer slurry, for growth and sporulation of Bacillus thuringiensis kurstaki was studied in baffled shake flasks. In a standard buffered medium with 2.5% (w/v) glucose and 1% (w/v) brewer's yeast extract, growth of B. t. kurstaki resulted in a low biomass production with considerable byproduct formation, including organic acids and a concomitant low medium pH, incomplete glucose utilization and marginal sporulation, whereas growth in the same medium with a commercial laboratory-grade yeast extract (Difco) resulted in a high biomass concentration, complete glucose utilization, relatively low levels of byproducts and complete sporulation (2.6 × 10⁹ spores/ml). When glucose was left out of the medium, however, growth parameters and sporulation were comparable for BYE and commercial yeast extract, but absolute biomass levels and spore counts were low. Iron was subsequently identified as a limiting factor in BYE. After addition of 3 mg iron sulphate/l, biomass formation in BYE-medium more than doubled, low byproduct formation was observed, and complete sporulation occurred (2.8 × 10⁹spores/ml). These data were slightly lower than those obtained in media with commercial yeast extract (3.6 × 10⁹spores/ml), which also benefited, but to a smaller extent, from addition of iron.     

富铬酵母的理化性质和氨基酸分析

用２００～３００ｎｍ的波长范围对富铬酵母及普通酵母的溶液进行紫外扫描,发现在λ_(２６０ｎｍ)处有一特征紫外吸收峰,富铬酵母细胞的铬含量与其吸收峰的光密度呈线性关系;在不同的温度和ｐＨ值条件下,通过紫外吸收峰的测定,富铬酵母溶液在酸性条件下稳定,在碱性条件下不稳定。溶液的λ_(２６０ｎｍ)紫外吸收峰随着温度的上升而升高。从氨基酸含量分析结果看,富铬酵母的谷氨酸、甘氨酸、组氨酸、丙氨酸和赖氨酸含量高于普通酵母,但其他氨基酸含量比普通酵母低。     

Repeated use of yeast biomass in ethanol fermentation of juniper berry sugars

Summary The object of this study was to establish the possibility of using the yeast biomass separated from the fermentation broth at the end of ethanol fermentation of juniper berry sugars as an inoculum in successive batch fermentation processes. A part of the fermentation broth (10% v/v) and a suspension of yeast biomass (separated from the same broth) into the water extract of juniper berries (2 g of wet yeast biomass per liter of water extract) were used as inocula. It was shown that the suspension of yeast biomass could be used as inoculum in successive batch processes without negative effects on the kinetics and ethanol yield, but with positive effects on the capacity and economy of the bioprocess. The addition of ammonium salts at optimum levels did not affect the final ethanol concentrations (4.3–4.4% v/v), but enhanced the specific rate of ethanol production, thus reducing the process duration by about five times.     

Effect of oils and fatty acids on molasses fermentation by distillers' yeast

Some vegetable oils, and the mixed fatty acids derived from them, stimulate sugar utilization and both the rate of alcohol production and yield of alcohol during molasses fermentation by yeast. The effect is particularly prominent during fermentation at a higher temperature of 40°C.     

Wort sugar uptake by brewer's yeast

Summary A culture of brewer's yeast,Saccharomyces cerevisiae (NCYC 240), maintained on Wickerham's MYGP medium, utilized the principal wort sugars sequentially in the order glucose-maltose-maltotriose, when inoculated into brewer's wort. A culture of the same strain maintained on brewer's wort utilized these three sugars simultaneously. Simultaneous utilization could be induced in MYGP-maintained cultures by successive sub-culture in brewer's wort, and appears to be the general rule of sugar uptake during wort fermentation under brewery conditions.     

海洋红酵母Y2发酵产类胡萝卜素条件的研究

目的 对海洋红酵母Y2高产类胡萝卜素的发酵条件进行优化.方法 在摇瓶条件下,研究培养基成分和培养条件对海洋红酵母Y2生长和类胡萝卜素合成的影响,同时进行海洋红酵母Y2发酵过程的动态分析.结果 海洋红酵母Y2优化培养基组合为葡萄糖45 g/L,蔗糖15 g/L,酵母粉5 g/L,蛋白胨2.5 g/L,磷酸二氢钾1 g/L,磷酸二氢钠3 g/L,硫酸镁7.5 g/L,氯化钾3 g/L,氯化钠5 g/L.最适培养参数为:温度20℃,培养基初始pH为5,接种量为10％,250 mL摇瓶装液量为10～50 mL.类胡萝卜素的合成主要集中在对数生长期和稳定期.海洋红酵母Y2最适收获时间为72 h.种龄以36 h为宜.结论 利用优化培养基,在最适条件下培养海洋红酵母Y2,类胡萝卜素产量达到4.97 mg/L,比基础培养基提高了60.32％.     

Ion exchange purification of G6PDH from unclarified yeast cell homogenates using expanded bed adsorption

The use of expanded beds of STREAMLINE ion exchange adsorbents for the direct extraction of an intracellular enzyme glucose-6-phosphate dehydrogenase (G6PDH) from unclarified yeast cell homogenates has been investigated. It has been demonstrated that such crude feedstocks can be applied to the bed without prior clarification steps. The purification of G6PDH from an unclarified yeast homogenate was chosen as a model system containing the typical features of a direct extraction technique. Optimal conditions for the purification were determined in small scale, packed bed experiments conducted with clarified homogenates. Results from these experiments were used to develop a preparative scale separation of G6PDH in a STREAMLINE 50 EBA apparatus. The use of an on-line rotameter for measuring and controlling the height of the expanded bed when operated in highly turbid feedstocks was demonstrated. STREAMLINE DEAE has been shown to be successful in achieving isolation of G6PDH from an unclarified homogenate with a purification factor of 12 and yield of 98% in a single step process. This ion exchange adsorbent is readily cleaned using simple cleaning-in-place procedures without affecting either adsorption or the bed expansion properties of the adsorbent after many cycles of operation. The ability of combining clarification, capture, and purification in a single step will greatly simplify downstream processing flowsheets and reduce the costs of protein purification. (c) 1996 John Wiley & Sons, Inc.     

Baker's yeast production from molasses/cheese whey mixtures

Partial substitution of sugarcane molasses by cheese whey in the fed-batch production of baker's yeast was evaluated. A sugar feeding profile based in a commercial process and different modes of addition of -galactosidase were used. Molasses substitution of 46%, in terms of sugar fed to the bioreactor, was reached and no significant differences in biomass volumetric productivity, by-products yields, and baking quality were observed. However the biomass yield was 6% lower.     

Theanine production by coupled fermentation with energy transfer employing Pseudomonas taetrolens Y-30 glutamine synthetase and baker's yeast cells

Theanine was formed from glutamic acid and ethylamine by coupling the reaction of glutamine synthetase (GS) of Pseudomonas taetrolens Y-30 with sugar fermentation of baker's yeast cells as an ATP-regeneration system. Theanine formation was stimulated by the addition of Mn2+ to the mixture for the coupling. The addition of Mg2+ was less effective. In a mixture containing a larger amount of yeast cells with a fixed level of GS, glucose (the energy source) was consumed rapidly, resulting in a decrease in the final yield of theanine. On the other hand, an increase in GS amounts increased theanine formation in a mixture with a fixed amount of yeast cells. High concentrations of ethylamine enhanced theanine formation whereas inhibited yeast fermentation of sugar and the two contrary effects of ethylamine caused a high yield of theanine based on glucose consumed. In an improved reaction mixture containing 200 mM sodium glutamate, 1,200 mM ethylamine, 300 mM glucose, 50 mM potassium phosphate buffer (pH 7.0), 5 mM MnCl2, 5 mM AMP, 100 units/ml GS, and 60 mg/ml yeast cells, approximately 170 mM theanine was formed in 48 h.     

Expression of GAI gene and disruption of PEP4 gene in an industrial brewer's yeast strain

Aims:  Construction of an industrial brewer's yeast strain, which could improve foam stability and reduce calorific values of beer.
Methods and Results:  An industrial brewer's yeast strain (Ts-10) was constructed by integrating glucoamylase encoding gene GAI amplified from Saccharomycopsis fibuligera by PCR into the locus of proteinase A (PrA) gene ( PEP4 ). The resulting recombinant strain identified by PCR could grow on YNB minimal medium plate with starch as sole carbon source. Its highest GAI activity was 91·69 U ml⁻¹, but it had no PrA activity. The real extract was reduced by 21·07% and the main residual maltotriose content was reduced by 14% in wort fermented with the recombinants strain. Its foam retention in beer was higher 39 s and the contents of potential off-flavour compounds, such as diacetyl, pentanedione and acetaldehyde were lowered by 16%, 13% and 14%, respectively, as compared with the industrial brewer's yeast YSF-5.
Conclusions:  An industrial brewer's yeast strain was constructed by introducing GAI gene and disrupting PEP4 gene.
Significance and Impact of the Study:  The recombinant strain (Ts-10) had better foam performance and mouthfeel in addition to low-calories values. It was free of heterologous DNA sequences and drug-resistance genes and could be safely used in beer production.     

Applications of yeast flocculation in biotechnological processes

A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects—the basics of yeast flocculation, the development of “new” flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous β-galactosidase production using a recombinant flocculentSaccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculation bioreactors and discussing potential new uses of these systems.     

啤酒酵母的开发应用及其制代可可粉的研究

本文介绍了啤酒酵母营养的成分和医用价值,概述了国内外开发应用啤酒酵母的研究功态,研究了用啤酒酵母加工代可可粉的工艺路线和方法,同时回收了啤酒,测定了代可可粉的营养成分,并用之加工巧克力和可可饮料,结果表明,其效果与天然可可粉类同。     

啤酒生产酵母全循环新工艺的研究

以改善浅色啤酒质量,降低生产成本,提高经济效益的目的,提出一种新颖的浅色啤酒的酿造方法。采用煮－浸法糖化工工艺及５０％麦芽和５０％大米作为啤酒酿造的原料及辅料。在糖化过程中添加啤酒酵母提取物作为补充氮源,不仅使所酿造的成品啤酒色泽浅,口味淡爽,纯正,泡沫洁白细腻,持久挂杯,而且具有较显著的经济效益和社会效益。     

Functional enrichment of mannanase-treated spent brewer yeast

In this study, the effect of Bacillus amyloliquefaciens-produced β-mannanase on the nutrient diffusion (release) and antioxidant activity of spent brewer yeast (SBY) was investigated. Three pretreatments were performed: (1) autolysis at 50°C for 24?h; (2) autolysis at 50°C for 24?h, with the addition of β-mannanase during the autolysis; (3) autolysis at 50°C for 24?h, and the β-mannanase was added for another 12?h treatment. The pretreatments with the addition of β-mannanase caused significant cell wall degradation, markedly increased the yield of SBY extracts. More importantly, this study found that polysaccharides were degraded to be oligosaccharides with a considerable reduction in molecular weights. Meanwhile, pretreatment with the enzyme also exhibited a higher antioxidant activity in SBY extract compared to autolysis itself. The current study indicated that pretreatment (3) had a better effect than pretreatment (2) in terms of improving in antioxidant activity in SBY extract. These improved characteristics of SBY extracts isolated through enzymatic treatment appear to show promising features for their prospective use as natural functional agents.     

Effects of yeast extract and glucose on xanthan production and cell growth in batch culture of Xanthomonas campestris

Although available kinetic data provide a useful insight into the effects of medium composition on xanthan production by Xanthomonas campestris, they cannot account for the synergetic effects of carbon (glucose) and nitrogen (yeast extract) substrates on cell growth and xanthan production. In this work, we studied the effects of the glucose/yeast-extract ratio (G/YE) in the medium on cell growth and xanthan production in various operating modes, including batch, two-stage batch, and fed-batch fermentations. In general, both the xanthan yield and specific production rate increased with increasing G/YE in the medium, but the cell yield and specific growth rate decreased as G/YE increased. A two-stage batch fermentation with a G/YE shift from an initial low level (2.5% glucose/0.3% yeast extract) to a high level (5.0% glucose/0.3% yeast extract) at the end of the exponential growth phase was found to be preferable for xanthan production. This two-stage fermentation design both provided fast cell growth and gave a high xanthan yield and xanthan production rate. In contrast, fed-batch fermentation with intermittent additions of glucose to the fermentor during the stationary phase was not favorable for xanthan production because of the relatively low G/YE resulting in low xanthan production rate and yield. It is also important to use a moderately high yeast extract concentration in the medium in order to reach a high cell density before the culture enters the stationary phase. A high cell density is also important to the overall xanthan production rate. Received: 30 September 1996 / Received revision: 21 January 1997 / Accepted: 10 February 1997     

Simultaneous saccharification and fermentation of cellulose for lactic acid production

Lactic acid production from α-cellulose by simultaneous saccharification and fermentation (SSF) was studied. The cellulose was converted in a batch SSF using cellulase enzyme Cytolase CL to produce glucose sugar andLactobacillus delbrueckii to ferment the glucose to lactic acid. The effects of temperature, pH, yeast extract loading, and lactic acid inhibition were studied to determine the optimum conditions for the batch processing. Cellulose was converted efficiently to lactic acid, and enzymatic hydrolysis was the rate controlling step in the SSF. The highest conversion rate was obtained at 46°C and pH 5.0. The observed yield of lactic acid from α-cellulose was 0.90 at 72 hours. The optimum pH of the SSF was coincident with that of enzymatic hydrolysis. The optimum temperature of the SSF was chosen as the highest temperature the microorganism could withstand. The optimum yeast extract loading was found to be 2.5 g/L. Lactic acid was observed to be inhibitory to the microorganisms’ activity.     

Recombinant brewer's yeast strains suitable for accelerated brewing.

Four brewer's yeast strains carrying the alpha-ald gene of Klebsiella terrigena (ex. Aerobacter aerogenes) or of Enterobacter aerogenes on autonomously replicating plasmids were constructed. The alpha-ald genes were linked either to the ADC1 promoter or to the PGK1 promoter of yeast Saccharomyces cerevisiae. In pilot scale brewing (50 l) with three of these recombinant yeasts the formation of diacetyl in beer was so low during fermentation that lagering was not required. All other brewing properties of the strains were unaffected and the quality of finished beers was as good as that of finished beer prepared with the control strain. The total process time of beer production could therefore be reduced to 2 weeks, in contrast to about 5 weeks required in the conventional process.     

纤维素酶水解啤酒糟的研究

研究了纤维素酶水解啤酒糟的适宜条件以及底物预处理方法对纤维素转化率和多糖水解率的影响。在适宜条件下,100g干啤酒糟可水解得10．8g还原糖。酶解液用于培养酵母菌提取麦角固醇,残渣是生产含菌体蛋白饲料的原料。     

Magnesium limitation and its role in apparent toxicity of ethanol during yeast fermentation

The rate of ethanol production per milligram of cell protein begins to decline in the early stage of batch fermentation before high concentrations of ethanol have accumulated. In yeast extract-peptone medium (20% glucose), this initial decline appears to be related to growth and to result in part from a nutrient deficiency. The addition of yeast extract, peptone, and ashed preparations of these restored the ability of glucose-reconstituted medium (in which cells had been previously grown) to support vigorous growth. Magnesium was identified as the active component. Supplementing fermentations with 0.5 mM magnesium prolonged exponential growth, resulting in increased yeast cell mass. The addition of magnesium also reduced the decline in fermentative activity (micromoles of CO2 evolved per hour per milligram of protein) during the completion of batch fermentations. These two effects reduced the time required for the conversion of 20% glucose into ethanol by 1/3 with no measurable loss in ethanol yield (98% of theoretical maximum yield). It is possible that some of the reported beneficial effects of complex nutrients (soy flour and yeast extract) for ethanol production also result from the correction of a simple inorganic ion deficiency, such as magnesium.