Downstream process for the production of yeast extract using brewer's yeast cells

A downstream process was developed for the production of yeast extract from brewer's yeast cells. Various downstream processing conditions including clarification, debittering, and the Maillard reaction were considered in the development of the process. This simple and economic clarification process used flocculating agents, specifically calcium chloride (1%). After the clarification step, a Maillard reaction is initiated as a flavor-enhancing step. By investigating the effects of several operation parameters, including the type of sugar added, sugar dosage, glycine addition, and temperature, on the degree of browning (DB), glucose addition and reaction temperature were found to have significant effects on DB. A synthetic adsorption resin (HP20) was used for the debittering process, which induced a compositional change of the hydrophobic amino acids in the yeast hydrolysate, thereby reducing the bitter taste. The overall dry matter yield and protein yield for the entire process, including the downstream process proposed for the production of brewer's yeast extract were 50 and 50%, respectively.     

Dietary autolysed yeast modulates blood profiles,small intestinal morphology and caecal microbiota of weaning pigs

Yeast products are potential feed additives due to their beneficial effects on gut health. Thus, we verified the potential impacts of autolysed yeast (AY) on growth performance, blood profiles, gut morphology and microbiota in weaning pigs. In total, 72 castrated, commercial, crossbred, weaning pigs were divided into three groups, with each group consisting of eight replicates with three piglets each. The experimental diets were as follows: 1) control diet (0% AY); 2) diet with 1.0% AY; 3) diet with 3.0% AY. For the overall period, using 1.0% AY in the diet seemed to improve the feed conversion ratio (P = 0.09); whereas, other productive performance parameters were not significantly affected by the supplementations. Using 1.0% AY in the diet significantly decreased the blood urea nitrogen and neutrophil/lymphocyte ratio (N/L ratio) but increased the eosinophil count (P < 0.05). Adding AY to the diet did not influence caecal microbial diversity; using 1.0% AY in the diet decreased the abundances of the phylum Actinobacteria, the class Coriobacteriia and the family Coriobacteriaceae (P < 0.05). At the genus level, an AY inclusion level of 1.0% reduced the abundances of Collinsella, Clostridium and Catenibacterium and increased that of Marvinbryantia (P < 0.05). Furthermore, the abundance of butyrate-producing bacteria seemed to be increased by AY supplementation (P = 0.06). Pearson’s correlation coefficient (r) analysis revealed that AY intake was negatively associated with the abundance of pathogens of the genera Dorea (r = ?0.84; P = 0.03) and Catenibacterium (r = ?0.80; P = 0.04). This indicates that AY intake potentially reduces the population of some pathogenic bacteria at family level. Thus, using an appropriate AY inclusion level (1.0%) seemed to improve the feed use of postweaning pigs and clearly improved their small intestinal morphology, blood profiles and caecal microbiota.     

Effect of yeast extract on speciation and bioavailability of nickel and cobalt in anaerobic bioreactors

The speciation of metals plays an important role in their bioavailability. In the case of anaerobic reactors for the treatment of wastewaters, the ubiquitous presence of sulfide leads to extensive precipitation of metals like nickel and cobalt, which are essential for the metabolism of the anaerobic microorganisms that carry out the mineralization of the pollutants present in the wastewater. In practice, nickel, cobalt, and iron are added in excessive amounts to full-scale installations. This study is concerned with the complexation of nickel and cobalt with yeast extract and its effect on the biogas production by methanogenic biomass. Adsorptive stripping voltammetry (AdSV) was used to get information about the stability and complexing capacity of the metal-yeast extract complexes formed. Nickel and cobalt form relatively strong organic complexes with yeast extract. The bioavailability of these essential metals in anaerobic batch reactors was dramatically increased by the addition of yeast extract. This is due to the formation of dissolved bioavailable complexes, which favors the dissolution of metals from their sulfides. Trace doses of yeast extract may be effective in keeping additions of essential metals to anaerobic reactors at a minimum.     

酵母浸出粉在奈瑟脑膜炎双球菌培养中的比较观察

目的探索A群,C群奈瑟脑膜炎双球菌(简称流脑)多糖疫苗生产中奈瑟脑膜炎双球菌培养的最适培养基。方法在培养基配制中用增减酵母浸出粉的方法制备相应的培养基,8h收菌,通过菌体的收获量并参考多糖量来确定较好的培养基配比。结果不同培养基用于A群、C群奈瑟脑膜炎菌培养8h后均有收获,其中2号培养基(酵母浸出粉)培养的菌体的浓度明显高于1号和3号培养基,它们之间有显著性差异(P<0.05)此种培养基能提高奈瑟脑膜炎双球菌的产量。结论添加酵母浸出粉的培养基可作为A群、C群奈瑟脑膜炎双球菌培养的最适培养基。     

High pressure homogenization inactivation of Escherichia coli and Staphylococcus aureus in phosphate buffered saline,milk and apple juice

High pressure homogenization (HPH) offers new opportunities for food pasteurization/sterilization. Escherichia coli and Staphylococcus aureus suspended in phosphate buffered saline (PBS) buffer, milk and apple juice at initial concentration of ~10⁶ log₁₀ CFU per ml were subjected to HPH treatments up to 200 MPa with inlet temperatures at 4–40°C. After HPH at 200 MPa with the inlet temperature at 40°C, the count of E. coli suspended in PBS, milk and apple juice reduced by 3·42, 3·67 and 3·19 log₁₀ CFU per ml respectively while the count of S. aureus decreased by 2·21, 1·02 and 2·33 log₁₀ CFU per ml respectively suggesting that S. aureus was more resistant. The inactivation data were well fitted by the polynomial equation. Milk could provide a protective effect for S. aureus against HPH. After HPH at 200 MPa with the inlet temperature at 20°C, the cell structure of E. coli was destroyed, while no obvious damages were found for S. aureus.     

Effect of yeast extract on growth kinetics during aerobic biodegradation of chlorobenzoic acids

The Monod or Andrews kinetic parameters describing the growth of Pseudomonas sp. CPE2 strain on 2,5-dich!orobenzoic acid and 2-chlorobenzoic acid, and Al-caligenes sp. CPE3 strain on 3,4-dichlorobenzoic acid, 4-chlorobenzoic acid, and 3-chlorobenzoic acid were determined from batch and continuous growth experiments conducted in the presence or absence of yeast extract (50 mg/L). Strain CPE2 displayed inhibitory growth kinetics in the absence of yeast extract and a noninhibitory kinetics in the presence of yeast extract. Similar results were obtained for CPE3. The presence of yeast extract also resulted in a significant increase in the affinity of the strains for the chlorobenzoic acids they degraded. (c) 1995 John Wiley & Sons, Inc.     

Hydrostatic pressure is like high temperature and oxidative stress in the damage it causes to yeast

Abstract A comparison of barotolerance, thermotolerance and oxygen tolerance was made under different physiological conditions, such as heat shocked and recovered state, different growth phases and changes of physiological conditions by mutations. The three kinds of tolerance showed similar features under different physiological conditions. We suggest that the damage caused by hydrostatic pressure may be essentially the same as that due to high temperature and oxidative stress in yeast.     

High gas pressure effects on yeast

Dried microorganisms are particularly resistant to high hydrostatic pressure effects. However, exposure to high pressures of nitrogen proved to be effective in inactivating dried yeasts. In this study, we tried to elucidate this mechanism on Saccharomyces cerevisiae. High-pressure treatments were performed using different inert gases at 150 MPa and 25 degrees C with holding time values up to 12 months. The influence of cell hydration was also investigated. For fully hydrated cells, pressurized gases had little specific effect: cell inactivation was mainly due to compression effects. However, dried cells were sensitive to high pressure of gases. In this latter case, two inactivation kinetics were observed. For holding time up to 1 h, the inactivation rate increased to 4 log and was linked to a loss of membrane integrity and the presence of damage on the cell wall. In such case cell inactivation would be due to gas sorption and desorption phenomena which would rupture dried cells during a fast pressure release. Gas sorption would occur in cell lipid phases. For longer holding times, the inactivation rate increased more slightly due to compression effects and/or to a slower gas sorption. Water therefore played a key role in cell sensitivity to fast gas pressure release. Two hypotheses were proposed to explain this phenomenon: the rigidity of vitrified dried cells and the presence of glassy solid phases which would favor intracellular gas expansion. Our results showed that dried microorganisms can be ruptured and inactivated by a fast pressure release with gases.     

A fast and simple approach to optimize the unit operation high pressure homogenization - a case study for a soluble therapeutic protein in E. coli

Escherichia coli is one of the most commonly used host organisms for the production of recombinant biopharmaceuticals. E. coli is usually characterized by fast growth on cheap media and high productivity, but one drawback is its intracellular product formation. Product recovery from E. coli bioprocesses requires tedious downstream processing (DSP). A typical E. coli DSP for an intracellular product starts with a cell disruption step to access the product. Different methods exist, but a scalable process is usually achieved by high pressure homogenization (HPH). The protocols for HPH are often applied universally without adapting them to the recombinant product, even though HPH can affect product quantity and quality. Based on our previous study on cell disruption efficiency, we aimed at screening operational conditions to maximize not only product quantity, but also product quality of a soluble therapeutic protein expressed in E. coli. We screened for critical process parameters (CPPs) using a multivariate approach (design of experiments; DoE) during HPH to maximize product titer and achieve sufficient product quality, based on predefined critical quality attributes (CQAs). In this case study, we were able to gain valuable knowledge on the efficiency of HPH on E. coli cell disruption, product release and its impact on CQAs. Our results show that HPH is a key unit operation that has to be optimized for each product.     

胃蛋白酶和木瓜蛋白酶水解核桃蛋白工艺研究

以核桃粕为原料,以水解度为考察指标,研究胃蛋白酶和木瓜蛋白酶对核桃蛋白的水解作用.结果表明：木瓜蛋白酶为酶解核桃蛋白最佳酶,其最佳酶解条件为底物浓度4％、酶质量分数5％、酶解 pH 值为7．0、酶解温度50℃、酶解时间4 h;在该条件下,木瓜蛋白酶酶解核桃蛋白水解度可高于25．76％.     

Effect of calcium and other metal ions on pectic activities from autolysed cultures of Alternaria alternata

Abstract The effect of increasing concentrations of Ca²⁺, Mg²⁺, Cu²⁺, Zn²⁺, Na⁺ and EDTA on the pectic enzymic activities (polymethylgalacturonase, endopectinase and pectin-lyase) present in the autolytic complex from Alternaria alternata has been studied. In all cases the divalent metal ions and EDTA produced an increased inhibition correlated with increasing concentration of each ion. An opposite effect was shown by the Na⁺ ion, which produced an increase in pectic enzymic activities, principally at low concentrations.     

Toward consistent and productive complex media for industrial fermentations: studies on yeast extract for a recombinant yeast fermentation process

Yeast extract (YE) is commonly used as a key component in the complex media for industrial fermentations. However, the lot-to-lot variation of this raw material frequently requires extensive "use testing" of many lots to identify only the few that support desired fermentation performance. Through extensive fermentation studies and chemical analyses, we have identified adenine and two metabolizable carbon sources, trehalose and lactate, as the principle components in YE that affect the production of a recombinant protein antigen by a yeast strain. Adenine is required for culture growth and the relationship between biomass and measured adenine can be expressed by a Michaelis-Menten model, while the slowly metabolized trehalose serves to maintain the energy supply to the continued antigen synthesis. The rapidly utilized lactate exerts an indirect positive effect by sparing some of the accumulated ethanol from being consumed for growth to being utilized in the product formation. The effects of these YE components are mutually dependent. Based on the database generated from 40 lots at laboratory scale, a relatively high level of carbon sources in YE (trehalose plus lactate, >9.5% w/w) and an intermediate level of adenine (0.14-0.24% w/w) appear to be the minimal requirement of a good lot for this recombinant yeast fermentation. Many poor lots were improved in lab fermenters by rational supplementation of trehalose, lactate, or adenine to compensate for their insufficiencies. At the large production scale, predictions based on adenine and trehalose/lactate contents in various YE lots used correlated reasonably well with culture growth and antigen yield, illustrating the feasibility of such a simple chemical/biochemical analysis as a rapid and reliable initial screening tool. Without incurring any compositional change to an established manufacturing medium, this study demonstrates an effective approach to achieve consistency in fermentations employing complex nutrients and to improve fermentation productivities supported by suboptimal lots of raw material.     

Influence of yeast extract concentrationon batch cultures of Lactobacillus helveticus: growth and production coupling

Lactobacillus helveticus was cultivated batchwise on whey permeate supplemented with 2–30g yeast extract l^-1. For low supplementation levels, growth was slow, and a long maintenance phase in the absence of any cell autolysis was observed. When the yeast extract concentration was increased, growth time increased slightly; under these circumstances, a large increase of maximum biomass concentration was recorded, indicating a clear nitrogen source starvation at low supplement concentration. At the same time the length of the maintenance phase decreased; for a 30gl^-1 supplementation, it vanished and a death phase was observed instead: this behaviour highlighted a shift from nitrogen to carbon source starvation. For more and more supplemented media, the final lactic acid concentration was constant, while fermentation time decreased significantly; a growth-associated production phase was followed by a maintenance production phase. The part played by the former mecha nism was under tight control of yeast extract concentration: for 30gl^-1 supplementation, 83% of total lactic acid was provided by a growth-linked production process.     

Effect of high hydrostatic pressure on four genotypes of F-specific RNA bacteriophages

AIMS: The pressure responses of four genotypes of F-specific RNA bacteriophages, f2, GA, Qbeta and SP, were evaluated with respect to pressure magnitude, treatment temperature and suspending medium. METHOD AND RESULTS: The pressure responses were studied with respect to pressure magnitude (350 to 600 MPa), treatment temperature (-10 to 50 degrees C) and suspending media. Phages f2 and GA had much higher pressure resistances than Qbeta and SP. Pressure resistances of Qbeta and SP were enhanced with increase in salt concentrations in the range of 350 to 600 MPa from -10 to 50 degrees C in PBS. Qbeta and SP had greater pressure resistances when suspended in phosphate-buffered saline (PBS) with added glucose (5%, w/w), UHT whole milk and Dulbecco's Modified Eagle's Medium plus 10% fetal bovine sera than they did in PBS. Two surfactants, sucrose laurate and monolaurin, and one chelating agent, ethylenediamine tetraacetic acid (EDTA), increased the pressure resistance of Qbeta and SP, but had modest effect on either f2 or GA. CONCLUSIONS: Four representative F-specific RNA bacteriophages, f2 (serotype I), GA (serotype II), Qbeta (serotype III) and SP (serotype IV) showed different resistances to hydrostatic pressure in the range of 350-600 MPa. Significance and Impact of the Study: This study screened for practical surrogates of HAV for validation of commercial high hydrostatic pressure processing.     

Ammonia production from yeast extract and its effect on growth of the hyperthermophilic archaeonSulfolobus solfataricus

Utilization of yeast extract and formation of byproduct metabolite were investigated for hyperthermophilic archaeonSulfolobus solfataricus (DSM 1617). In both batch and fed-batch cultivations ofS. solfataricus, maximal cell density, NH₄ ⁺ ion production and pH change were highly dependent on the ratio of yeast extract to glucose in the medium. Variation of NH₄ ⁺ ion level was identified as a major cause of pH change during cultivation, and acidification of culture broth was attributed to consumption of NH₄ ⁺ ions rather than formation of acid byproducts. It was also observed that increase of NH₄ ⁺ ion concentrations in the medium resulted in greater degree of growth inhibition.     

酵母菌体在食品中开发与应用

介绍以酵母菌体为原料制取食用蛋白、酵母精及增鲜剂，并对各制品在食品中的应用进行了阐述，旨在推动酵母菌体开发与利用的进程。     

Post-irradiation modification of radiation-induced heteroallelic reversion in diploid yeast: Effect of nutrient broth

The effect of post-irradiation growth in complete rich medium on the expression of the reversion to arginine-independence induced by gamma and alpha radiation in a heteroallelic diploid yeast strain (Saccharomyces cerevisiae BZ34) has been studied. During the post-irradiation treatment the reversion frequency increased, reached a peak at about 90 min and decreased thereafter reaching a constant value for treatment periods exceeding 6 h. As determined by the increase in number of budding cells, extensive DNA synthesis took place in cells incubated only in the nutrient medium and not in the omission medium. Hence the observed increase in the reversion frequency is explained on the basis that post-irradiation DNA synthesis is necessary for the expression of gene conversion. The decrease in the reversion frequency for continued treatment with yeast extract, peptone, dextrose (YEPD) is related to the fact that only one daughter of the post-irradiation first cell division is a revertant.The broth effect was not lost when the irradiated cells were first incubated for 90 min in arginine-less medium and then transferred to the broth. Similarly, the broth effect persisted even at doses high enough to induce considerable division delay. These results suggest that the radiation-induced pre-conversional lesions are not susceptible to repair by alternative pathways.     

中温淡盐法制备低核酸酵母抽提物的工艺研究

通过建立一种新的酵母核酸中温淡盐去除法,成功研制出品质优良的低核酸酵母抽提物。工艺简单,处理时间短,温度不超过70℃,可避免高温对酵母营养物质的破坏;该法氯化钠用量少,无需脱盐处理,易产业化及应用推广;研究制备的酵母抽提物,核酸去除量达70%以上,色浅、肉香味鲜,适合食品加工。同时,附加得到高提取率、高纯度的核酸。中温淡盐法去除酵母核酸的技术,扩展了酵母制品的应用领域,更为尿酸偏高及痛风人群食用安全营养的酵母食品提供了解决方案。     

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.