Fehlermöglichkeiten der Gelöstsauerstoffmessung beim Betrieb von Bioreaktoren

A review of errors, which can influence the measured data of electrochemical oxygen sensors (OS) in fermentation technique is presented. The specifities of various sensor constructions are pointed out. References are given for selection of sensors to application in fermenters. Influences on the measured values can take place by fermentation conditions, arrangement of sensors in the fermenter and composition of the fermentation broth. The oxygen measurement in the multi-phase system of fermentation fluids can cause a remarkable deviation of the measured values to the real ones. Practical hints are given for use, calibration and sterilization of OS. Restricting conditions for measurements with this sensors are enumerated.     

Solid-state fermentation--are there any biotechnological advantages?

Solid-state fermentation (SSF) has developed in eastern countries over many centuries, and has enjoyed broad application in these regions to date. By contrast, in western countries the technique had to compete with classical submerged fermentation and, because of the increasing pressure of rationalisation and standardisation, it has been widely superseded by classical submerged fermentation since the 1940s. This is mainly because of problems in engineering that appear when scaling up this technique. However, there are several advantages of SSF, for example high productivities, extended stability of products and low production costs, which say much about such an intensive biotechnological application. With increasing progress and application of rational methods in engineering, SSF will achieve higher levels in standardisation and reproducibility in the future. This can make SSF the preferred technique for special fields of application such as the production of enzymes and food.     

Monitoring of Lactobacillus fermentation process by using ion chromatography with a series piezoelectric quartz crystal detector

A new method monitoring Lactobacillus fermentation process, which combines ion chromatography (IC) with a series piezoelectric quartz crystal (SPQC) technique, is presented in this paper. Monitoring of the fermentation process was realized by examining the rate of production of lactic acid. An automatic membrane dialyser was used for the pretreatment of the sample in on-line monitoring. A mixture of p-hydroxybenzoic acid and N,N-diethylethanolamine was adopted as mobile phase and its flow rate was 0.8 ml/min. The effects of some fermentation conditions were also discussed in detail. Accordingly, the optimal fermentation conditions were obtained. This method is simple and convenient while the results obtained are accurate and reliable.     

Cellulosic Butanol (ABE) Biofuel Production from Sweet Sorghum Bagasse (SSB): Impact of Hot Water Pretreatment and Solid Loadings on Fermentation Employing <Emphasis Type="Italic">Clostridium beijerinckii</Emphasis> P260

A novel butanol fermentation process was developed in which sweet sorghum bagasse (SSB) was pretreated using liquid hot water (LHW) pretreatment technique followed by enzymatic hydrolysis and butanol (acetone butanol ethanol (ABE)) fermentation. A pretreatment temperature of 200 °C resulted in the generation of a hydrolyzate that inhibited butanol fermentation. When SSB pretreatment temperature was decreased to 190 °C (0-min holding time), the hydrolyzate was successfully fermented without inhibition and an ABE productivity of 0.51 g L^?1 h^?1 was achieved which is comparable to the 0.49 g L^?1 h^?1 observed in the control fermentation where glucose was used as a feedstock. These results are based on the use of 86 g L^?1 SSB solid loadings in the pretreatment reactors. We were also able to increase SSB solid loadings from 120 to 200 g L^?1 in the pretreatment step (190 °C) followed by hydrolysis and butanol fermentation. As pretreatment solid loadings increased, ABE yield remained in the range of 0.38–0.46. In these studies, a maximum ABE concentration of 16.88 g L^?1 was achieved. Using the LHW pretreatment technique, 88.40–96.00 % of polymeric sugars (cellulose + hemicellulose) were released in the SSB hydrolyzate. The LHW pretreatment technique does not require chemical additions and is environmentally friendly, and the hydrolyzate can be used successfully for butanol fermentation.     

A reliable method for detecting the intracellular accumulation of fermentation products: Application to intracellular ethanol analysis

Summary Intracellular ethanol concentrations inSaccharomyces cerevisiae were measured using a high cell density fermentation technique which permits samples to be fixed for analysis in less than two seconds. No accumulation of intracellular ethanol was detected, regardless of the stage of fermentation. The technique eliminates artifacts associated with concentrating the cell sample and provides a reliable means for detecting whether other fermentation products accumulate intracellularly.     

盐析萃取生物基化学品的研究进展

廉价生物质的生物炼制研究主要集中在菌种和发酵方面,对下游分离研究较少。廉价生物质资源的利用导致发酵液中引入更多杂质,成分较单糖发酵更复杂,致使生物基化学品的下游分离过程成为其工业化生产亟需解决的关键问题。文中介绍了一种基于两相分配差异分离亲水性生物基化学品的盐析萃取技术及其在生物基化学品分离方面的应用,重点阐述了短链醇和盐对双水相形成的影响,并对1,3-丙二醇、2,3-丁二醇、乙偶姻、乳酸等的盐析萃取研究进展进行了总结和展望。盐析萃取技术可有效地回收发酵液中的小分子亲水性产品,同时除去大多数的杂质 (细胞和蛋白质等),在生物基化学品的分离过程中将是一种有前景的分离技术。     

Semi-continuous ethanolic fermentation using a novel yeast settling and recycle technique

Summary A novel technique for the rapid settling of yeast cells is outlined. An inert, high density powder is added to a yeast suspension and the pH of the suspension is switched rapidly from 4.5 (fermentation pH) to 8.0. Large, rapid settling flocs of yeast are formed immediately. This technique has been applied to the recycling of yeast from an ethanolic fermentation.     

大肠杆菌酪氨酸转运系统基因敲除对酪氨酸生产的影响

酪氨酸是三大芳香族氨基酸之一,广泛用于食品、医药和化工等领域。转运系统工程为代谢工程改造大肠杆菌选育酪氨酸生产菌株提供了一种重要的研究策略。大肠杆菌中酪氨酸胞内转运主要通过aroP和tyrP基因编码的通透酶进行调控。以酪氨酸生产菌株HGXP为出发菌株,利用CRISPR-Cas9技术成功构建了aroP和tyrP基因敲除菌,并通过发酵试验考察了调节转运系统对酪氨酸生产的影响。发酵结果表明,aroP和tyrP基因敲除菌酪氨酸产量分别达到3.74 g/L和3.45 g/L,较出发菌株酪氨酸产量分别提高了19%和10%。对诱导温度进行了优化,结果表明38℃为最佳诱导温度。在3 L发酵罐上进行了补料分批发酵,aroP和tyrP基因敲除菌酪氨酸产量进一步提高至44.5 g/L和35.1 g/L,较出发菌株酪氨酸产量分别提高了57%和24%。研究结果对代谢工程强化大肠杆菌生产酪氨酸具有重要的参考价值。     

内生真菌产紫杉醇研究的回顾与展望

紫杉醇是一种高效、低毒、广谱的天然抗癌药物,是人类未来20年间最有效的抗癌药物之一。近年来的研究发现,从植物内生真菌中发酵生产紫杉醇被证明是解决药源问题的有效途径。本文概述了微生物发酵生产紫杉醇的研究进展,包括产紫杉醇内生真菌的分离、多样性和真菌产紫杉醇的优势,紫杉醇的提取和检测技术,同时对如何提高内生真菌紫杉醇产量进行了比较全面的综述。随着现代分子生物学技术和微生物发酵工程的发展,工业上大规模发酵生产紫杉醇将有望实现。     

柑橘果实加工废料资源化新技术研究

本文报道柑橘果实加工废料经直接压榨,并利用微生物发酵制取乙醇、橙皮甙及饲料等新技术。乙醇、橙皮甙产品经检验达A.R.级;饲料中脂肪、蛋白质含量分别比发酵前提高了175.7%和51.1%。制备全过程不产生污染,便于产业化。     

Extractive fermentation of lactic acid by immobilizedLactobacillus casei using ion—exchange resin

Summary A novel method of lactic acid fermentation byLactobacillus casei immobilized in Ca—alginate gels is described, in which an ion—exchange resin packed column is attached to a fermentor for separation of lactic acid from fermentative broth. The technique successfully alleviated the restriction imposed by lactic acid on bacterial growth and product formation. As compared to the conventional batch fermentation, the new fermentation technique enhanced the lactic acid productivity and sugar conversion rate from 0.328g/L·h and 88. 2% to 0.482g/L·h and 98.6%, respectively.     

On-line adaptive control of a fed-batch fermentation of Saccharomyces cerevisiae

The feasibility of applying an adaptive control technique to a fermentation process is investigated. The nonlinear, time-variant parameters of a fermentation process were estimated on-line as a series of linearized describing matrices. The matrices were used to update a suboptimal feedback law which controlled the process in real time over the linear region. Experiments were performed on a small-scale fully instrumented fermenter with the online, real-time adaptive control package. Results are presented for both single- and multivariable control, and indicate successful control of yeast cell growth.     

New concepts for rapid yeast settling. II. pH switching with an inert powder

A new technique is outlined for the rapid settling of yeast cells in fermentation media. The technique involved the addition of dense, inert particles (nickel powder) to a yeast suspension (Saccharomyces cerevisiae) at pH 4.5 and a rapid change of pH to 8.0-9.0. When the pH was changed large flocs formed immediately and settled rapidly, leaving a clear supernatant. On returning the pH to 4.5 the flocs were destroyed. This technique gave larger flocs and higher settling rates than the constant pH method, and much lower nickel/yeast ratios were required. Good flocculation also occurred in a fermentation medium. The technique was used to recycle yeast cells to a semicontinuous ethanol fermentation. Application of the technique to this and similar systems is discussed. The factors affecting yeast/inert powder flocculation are also discussed and a model is proposed to explain the observed experimental behavior for flocculation with a rapid change in pH.     

毕赤酵母工程菌高密度发酵的研究进展

近年来毕赤酵母已成为一种优越的异源蛋白表达系统。而提高目的蛋白的表达水平,高密度发酵已成为关键技术环节之一。我们从毕赤酵母工程菌的选择、培养基的优化设计,以及发酵工程过程控制等方面简要阐述毕赤酵母的高密度发酵,并提出了工程菌在高密度发酵过程中存在的问题。     

Oxygen consumption as a measure yeast starter culture vitality for the wine industry

Summary A simple technique for rapid determination of fermentation starters vitality which eliminates the need for determination of viable cells counts is described. The mathematical relationship between cell number and oxygen consumption of eight strains of Saccharomyces cerevisiae was studied. Results confirmed the possibility of utilizing a pO2 probe as an indicator of cell viability for fermentation starter.     

Effects on product formation in Lactococcus lactis 65.1 in continuous culture with complete cell recycling

Lactococcus lactis 65.1 was cultivated in a batch culture, which underwent starvation for 3 days, continuous culture and continuous culture with complete cell recycling. The objective was to study the product formation and intracellular protein pattern. Changes from homofermentative to heterofermentative metabolism were observed in continuous culture at the lower dilution rates as well as continuous cultures with complete cell recycling at a fixed dilution rate (D=0.4 h^–1). Changes in intracellular protein pattern were observed when starving the cells in a batch culture and also when recycling the cells in a continuous culture. Some changes were the same in these two cases. The data collected from these experiments show how the fermentation technique can affect the products of the microorganism being cultured and gives some interesting information on the complete cell recycling technique, which is of great interest in fermentation processes.     

Studies of Saccharomyces cerevisiae during fermentation--an in vivo electrokinetic investigation

An extensive in vivo study of the electrokinetic properties of six strains of the brewing yeast S. cerevisiae has been carried out. The yeasts were cultured under laboratory conditions. They were electrokinetically characterized by the electro-osmotic dipped cell technique, and data are presented as zeta-potentials. The effects of pH, fermentation time, successive fermentation cycles, and initial wort density have been established. The electrokinetic properties of an ale yeast which did not function correctly during commercial fermentation have also been examined. The results are discussed in the context of two controversial topics concerning the mechanism of yeast flocculation, the relative importance of yeast cell wall carboxyl and phosphate groups and the influence of wort components.     

By-product inhibition effects on ethanolic fermentation by Saccharomyces cerevisiae

Inhibition by secondary fermentation products may limit the ultimate productivity of new glucose to ethanol fermentation processes. New processes are under development whereby ethanol is selectively removed from the fermenting broth to eliminate ethanol inhibition effects. These processes can concentrate minor secondary products to the point where they become toxic to the yeast. Vacuum fermentation selectively concentrates nonvolatile products in the fermentation broth. Membrane fermentation systems may concentrate large molecules which are sterically blocked from membrane transport. Extractive fermentation systems, employing nonpolar solvents, may concentrate small organic acids. By-product production rates and inhibition levels in continuous fermentation with Saccharomyces cerevisiae have been determined for acetaldehyde, glycerol, formic, lactic, and acetic acids, 1-propanol, 2-methyl-1-butanol, and 2,3-butanediol to assess the potential effects of these by-products on new fermentation processes. Mechanisms are proposed for the various inhibition effects observed.     

A continuous fermentation technique for studying the kinetics of sugar uptake by baker's yeast

1. Solutions of glucose, maltose or other sugars were pumped at controlled rates into a yeast suspension and the extracellular sugar concentration was determined. The technique was especially suitable for studying the kinetics of fermentation at low rates of sugar utilization. At high rates the fermentation system was unstable. 2. Glucose fermentation was fitted by a model in which a diffusion barrier with first-order kinetics is interposed between the environment and the site of hexokinase. 3. Aerobic conditions affected the fermentation enzyme system but not the diffusion mechanism. 4. The kinetics of maltose fermentation at low rates approximated to those of the hydrolysis of maltose by the enzyme maltase, as studied in suspensions of broken cells.     

Solid-state (solid-substrate) food/beverage fermentations involving fungi

“Solid-substrate” fermentation developed in the Orient is a very useful fermentation method. It is presently used to produce a variety of foods, beverages and related products. Solid-substrate fermentation products utilizing fungi including soy sauce, miso and tempe, ontjom, sake, and bread have been produced for centuries at the home and village level. They are examples of economical methods of preserving and improving the flavor, texture and nutritive values of cereal/legume substrates. “Solid-substrate” fermentation is also applied to animal products such as milk to produce Roquefort and Camembert cheeses which diversify the food flavors available to man “Solid-substrate”fermentation has certain advantages. The substrate is concentrated; the product can be extracted with relatively small quantities of solvent; the product can be easily dehydrated; moisture level can be controlled favoring the desired organisms; enzyme concentration is generally higher than is submerged fermentation; product concentration is generally higher than in submerged cultures; it is the only technique that yields true mushroom fruiting bodies and it can be used not only for production of crude enzyme concentrates (koji) but also for raising the protein content of high starch substrates. It also can be used to increase the content of vitamins at low cost. Disadvantages of “solid-substrates”from the modern industrial processing view point are the greater difficulty of handling solid substrate and the greater difficulty of controlling the fermentation parameters, temperature, pH and oxygen, and rate of microbial growth compared with liquid submerged fermentations.