Very high gravity wort fermentation by immobilised yeast

Using calcium alginate-entrapped yeast, 24% (w/w) wort was successfully fermented within 8 days. This is half the time needed for fermentation by free yeast. The highest ethanol concentration obtained was 10.5% (v/v). When the original wort gravity was increased, the specific rate of ethanol production remained constant 0.16 g gh^–1 and the viability did not fall bellow 95% of living cells. Protection of cell against osmotic stress by gel matrix was also confirmed by trehalose measurement. The maximum intracellular trehalose content in calcium alginate-entrapped yeast was 3 times lower compared to free yeast at 30% (w/w) wort fermentation.     

一种固定化酵母细胞的复合载体——PVA-海藻酸盐凝胶

固定化细胞这一生物技术在近20年来发展迅速,已涉及食品、发酵、化工、医疗、生化等各领域.新型载体研制是发展固定化细胞技术的一个主导因素.PVA(聚乙烯醇)是近年来用于固定化细胞的一种新载体材料,但目前制备PVA凝胶的方法通常是在PVA溶液凝固后(或加入固化剂、乳化剂等让其凝固)手工切块.为了解决PVA载体的成球问题,简化制备方法,我们进行了这方面的研究.本文报道了用PVA和海藻酸钠制备的一种固定化细胞载体新配方,比较了几种常用载体及几种PVA载体的性能及用于乙醇发酵的实验结果.     

左旋麻黄素半生物合成的研究固定化酵母细胞生物合成L-PAC(L-苯基乙酰基甲醇)

本文对影响酵母菌生物合成L-PAC的主要因素进行了研究.结果表明酵母菌株Sc-5按2g鲜细胞/3.2%50ml海藻酸钠凝胶固定化后,所获固定化凝胶珠置于装有4倍体积反应液的容器中,在振荡频率220转/分、温度28-30℃、添加0.2%的Vc时,枇次生物合成L-PAC产量最高,达2.0g/L.     

固定化增殖酵母10m^3生物反应器连续生产酒精的研究

将经过驯化的酒精酵母,用海藻酸钙凝胶包埋法固定化,再用硫酸钴处理。在形成的凝胶中含有微量的甾醇和不饱和脂肪酸,以提高细胞的存活率。反应器系统由三个１０ｍ３稍加改造的普通发酵罐组成。用经过酸化处理的甜菜糖蜜为原料,采用逐步提高糖浓度的方法连续生产高浓度酒精,酒精含量为１０％（Ｖ／Ｖ）以上。发酵过程中通入微量无菌空气。酒精生产能力为５１．７５ｋｇ／ｍ３凝胶·ｈ－１,停留时间为１．８ｈ。反应器内形成的大量ＣＯ２通畅排出。反应器具有良好的操作稳定性。反应器容积生产强度为８．３ｋｇ／ｍ３·ｈ     

海藻酸铝固定化酵母生产高浓度酒精的研究

以海藻酸铝凝胶代替海藻酸钙,可使固定化酵母使用寿命显著延长,其海藻酸铝凝胶耐磷酸盐能力比海藻酸钙凝胶提高六倍以上。用海藻酸铝固定化增殖酵母进行分批发酵酒精,成熟醪中酒精含量由３．５％－９．０％（Ｖ／Ｖ）提高到１１．０％（Ｖ／Ｖ）左右。在１．１Ｌ的两个多层生物反应器中,装入海藻酸铝固定化增殖酵母,采用逐步提高糖浓度方法,进行连续发酵,成熟醪接收器中酒精浓度平均为１０．３％（Ｖ／Ｖ）,总糖利用率为９２．４％。     

耐高温酵母菌的筛选及特性

筛选得到1株在41 ℃可以良好生长的耐高温酵母菌,并对其特性及发酵性能进行研究.结果表明:该菌的最适生长pH为5.0,转速180 r/min,最高生长温度45 ℃.在41 ℃和初糖质量浓度为93.0 g/L的条件下发酵24 h后,乙醇质量浓度达到44.0 g/L,乙醇得率为93.0%.     

Continuous secondary fermentation using immobilised yeast

An immobilised yeast, two-stage reactor system was applied to laboratory fermentations of wort. The first stage, the primary fermentation process was carried out in an up-flow gas-lift bioreactor, and in the second stage, fermentation was processed in column reactors consisting of eight packed-beds filled with both yeasts entrapped in three different polysaccharide hydrogels (calcium alginate, calcium pectate, sodium carrageenan) and with free yeast in parallel. The residence time for the two-stage immobilised system varied from 74 to 108 h. © Rapid Science Ltd. 1998     

固定化生长酵母连续生产酒精的研究

采用固定化生长细胞方法,以柱式生物反应器连续发酵甜菜糖蜜酒精。酒精能力为39.45g/L凝胶/h,停留时间1.8小时。生物反应器具有良好的稳定性,连续工作50天,发酵醪酒精含量在8.5％(v/v)以上。系统研究了最适固定化条件,用L_(16)(4~5)正交试验确定了最佳发酵条件。     

Diffusivity of Cu2+ in calcium alginate gel beads

The diffusivity of Cu(2+) in calcium alginate beads calculated by the shrinking core model (SCM) was reevaluated in this work. The results obtained in this work were significantly different than those by the original authors. There were excellent agreements between the results obtained by the SCM in this work and those by the more rigorous linear absorption model (LAM) by the original authors. (c) 1994 John Wiley & Sons, Inc.     

Enhanced production of bioethanol and ultrastructural characteristics of reused Saccharomyces cerevisiae immobilized calcium alginate beads

Yeast immobilized on alginate beads produced a higher ethanol yield more rapidly than did free yeast cells under the same batch-fermentation conditions. The optimal fermentation conditions were 30 °C, pH 5.0, and 10% initial glucose concentration with 2% sodium alginate beads. The fermentation time using reused alginate beads was 10-14 h, whereas fresh beads took 24 h, and free cells took 36 h. All bead samples resulted in nearly a 100% ethanol yield, whereas the free cells resulted in an 88% yield. Transmission electron microscopy (TEM) showed that the shortened time and higher yield with the reused beads was due to a higher yeast population per bead as well as a higher porosity. The ultrastructure of calcium alginate beads and the alginate matrix structure known as the “egg-box” model were observed using TEM.     

Production of Food and Fodder Yeasts

Abstract

A decade or so ago, there was considerable interest in developing single cell protein production from raw materials. Many factors have influenced the development of fodder yeast technology, notably the biochemistry and physiology of the yeast.

It is shown that those considerations have led to the choice of a continuous fermentation technology.     

氨基化硅胶载体固定化纤维素酶的研究

用硅胶作载体,戊二醛作交联剂,制备了固定化的纤维素酶。对制备固定化纤维素酶的偶联剂浓度、pH、给酶量3个影响因素进行了研究,通过正交试验优化得出最佳的固定化条件:交联剂戊二醛浓度为1%,固定化pH值为5,固载量为每克载体100mg纤维素酶。     

Long-term incomplete xylose fermentation, after glucose exhaustion, with Candida shehatae co-immobilized with Saccharomyces cerevisiae

Saccharomyces cerevisiae and Candida shehatae were co-immobilized in an agar sheet which was introduced in an original two-chambered bioreactor asymmetrically fed in a batch mode with a mixture of glucose and xylose in a ratio of 35:15. The two sugars were consumed simultaneously. All glucose was fermented but only 20% of xylose. After incubation, yeast cells recovered from different areas of the agar sheet (close to, called Hi, and distant from, called Ho, the substrate chamber) were cultured as suspended cells in fresh culture medium provided with xylose or the sugar mixture. Xylose utilization by gel released Hi yeasts was significantly delayed compared to the Ho culture. Ethanol consumption by Hi yeasts in the two-substrate medium occurred after glucose exhaustion despite the presence of xylose. The waste medium resulting from incubation of the immobilized-cell structure inhibited xylose utilization by C. shehatae. Our results suggested that batch fermentation most probably favoured this incomplete xylose fermentation.     

Immobilization of cells by electrostatic droplet generation: a model system for potential application in medicine

The process of electrostatic extrusion as a method for cell immobilization was investigated that could be used for potential applications in medicine. An attempt was made to assess the effects of cell addition and polymer concentration on the overall entrapment procedure, ie, on each stage of immobilization: polymer-cell suspension rheological characteristics, electrostatic extrusion process, and the process ofgelation. The findings should contribute to a better understanding of polymer-cell interactions, which could be crucial in possible medical treatments. Alginate-yeast was used as a model system for carrier-cells. The electrostatic extrusion was considered as a complex two-phase flow system and the effects of cell and alginate concentrations on the resulting microbead size and uniformity were assessed. Under investigated conditions, microbeads 50-600 microm in diameter were produced and the increase in both alginate and cell concentrations resulted in larger microbeads with higher standard deviations in size. We attempted to rationalize the findings by rheological characterization of the cell-alginate suspensions. Rheological characterization revealed non-Newtonian, pseudoplastic behavior of cell-alginate suspensions with higher viscosities at higher alginate concentrations. However, the presence of cells even at high concentrations (5x10(8) and 1x10(9) cells/mL) did not significantly affect the rheological properties of Na-alginate solution. Lastly, we investigated the kinetics of alginate gelation with respect to the quantity of Ca2+ ions and cell presence. The gelation kinetics were examined under conditions of limited supply with Ca2+ ions, which can be essential for immobilization of highly sensitive mammalian cells that require minimal exposure to CaCl2 solution. The molar ratio of G units to Ca2+ ions of 3.8:1 provided complete crosslinking, while the increase in alginate concentration resulted in prolonged gelation times but higher strength of the resulting gel. The cell presence decreased the rate of network formation as well as the strength of the obtained Ca-alginate hydrogel.     

固定化乳酸乳球菌连续生产Nisin的研究

以海藻酸钙为材料 ,固定乳酸乳球菌 (Lactococcuslactissubsp .lactis)SM5 2 6 ,研究不同条件对Nisin合成的影响。结果表明 ,利用 2 %海藻酸钠在 1 0mmol LCaCl2 条件下 ,得到的固定化细胞颗粒稳定性较好 ,可维持 90h无破裂 ;在发酵过程中SYS3培养基中的无机盐成分尤其磷酸盐对固定化颗粒有破坏作用 ;用mSYS3培养基代替SYS3 ,通过 72h三批次循环的半连续培养 ,Nisin活性为 85 0IU mL ,无明显的细胞渗漏现象。连续化生产 70h ,Nisin活性达 1 1 5 0IU mL ,相当于游离细胞的发酵水平。     

Vitamin analysis with use of a yeast electrode

A vitamin B₁ (thiamin)-sensitive electrode has been devised by combining an oxygen electrode with a yeast-containing membrane. The assembly was used for assaying thiamin at concentrations down to 10^?11 gl^?1. The analytical procedure developed should allow the measurement of 10–20 samples per hour. The performance of the yeast electrode was improved when alginate membranes reinforced with a nylon network were used. An apparatus for preparing such membranes is described together with a magnetic membrane holder facilitating handling of membranes in combination with electrodes.     

Two-stage fermentation process for alginate production by Azotobacter vinelandii mutant altered in poly-β-hydroxybutyrate (PHB) synthesis

Aims:  A two-stage fermentation strategy, based on batch cultures conducted first under non-oxygen-limited conditions, and later under oxygen-limited conditions, was used to improve alginate production by Azotobacter vinelandii (AT6), a strain impaired in poly-β-hydroxybutyrate (PHB) production.
Methods and Results:  The use of sucrose as carbon source, as well as a high oxygen concentration (10%), allowed to obtain a maximum biomass concentration of 7·5 g l⁻¹ in the first stage of cultivation. In the second stage, the cultures were limited by oxygen (oxygen close to 0%) and fed with a sucrose solution at high concentration. Under those conditions, the growth rate decreased considerably and the cells used the carbon source mainly for alginate biosynthesis, obtaining a maximum concentration of 9·5 g l⁻¹, after 50 h of cultivation.
Conclusion:  Alginate concentration obtained from the AT6 strain was two times higher than that obtained using the wild-type strain (ATCC 9046) and was the highest reported in the literature. However, the mean molecular mass of the alginate produced in the second stage of the process by the mutant AT6 was lower (400 kDa) than the polymer molecular mass obtained from the cultures developed with the parental strain (950 kDa).
Significance and Impact of the Study:  The use of a mutant of A. vinelandii impaired in the PHB production in combination with a two-stage fermentation process could be a feasible strategy for the production of alginate at industrial level.