Production and characterization of the exopolysaccharide produced by <Emphasis Type="Italic">Paenibacillus jamilae</Emphasis> grown on olive mill-waste waters

Paenibacillus jamilae, a strain isolated from compost prepared with olive-mill wastewaters, produced an extracellular polysaccharide (EPS) when it was grown in a culture containing olive-mill waste waters (OMWW) as sole carbon and energy sources. Maximal EPS production in 100 mL batch-culture experiments (5.1 g L⁻¹) was reached with a concentration of 80% of OMWW as fermentation substrate (v/v). Although an inhibitory effect was observed on growth and EPS production when OMWW concentration was increased, an appreciable amount of EPS (2.7 g L⁻¹) was produced with undiluted OMWW. Sepharose CL-2B chromatography showed that the EPS presented two fractions, EPS I (>2000 kDa) and EPS II (500 kDa). Both fractions were characterized by GC-MS as two different acidic heteropolysaccharides containing glucose, galactose and mannose as the major components. The performed study made evident the possibility of using OMWW as substrate for the production of EPS by P. jamilae with a satisfactory yield.     

生物量浓度实时在线检测方法的研究

微生物的存在会改变发酵液的电特性,发酵液在无线电频率范围内的电容率增量是测量频率和生物量浓度的函数.基于对发酵液电容率分布的研究,提出了测量生物量浓度的新方法.用此方法不用取样就能对发酵液中的生物量进行实时在线测量,而且测得的是活的生物量浓度.制作的电极直接插入发酵器中并满足高温蒸气灭菌条件.此方法在生化制药、食品发酵、啤酒酿造、污水检测等工业领域里有很好的推广应用前景.     

A model of xylitol production by the yeast Candida mogii

Production of xylitol from xylose in batch fermentations of Candida mogii ATCC 18364 is discussed in the presence of glucose as the cosubstrate. Various initial ratios of glucose and xylose concentrations are assessed for their impact on yield and rate of production of xylitol. Supplementation with glucose at the beginning of the fermentation increased the specific growth rate, biomass yield and volumetric productivity of xylitol compared with fermentation that used xylose as the sole carbon source. A mathematical model is developed for eventual use in predicting the product formation rate and yield. The model parameters were estimated from experimental observations, using a genetic algorithm. Batch fermentations, which were carried out with xylose alone and a mixture of xylose and glucose, were used to validate the model. The model fitted well with the experimental data of cell growth, substrate consumption and xylitol production.     

Production of tannase by <Emphasis Type="Italic">Aspergillus niger</Emphasis> HA37 growing on tannic acid and Olive Mill Waste Waters

Production of tannase (tannin acyl hydrolase, EC 3.1.1.20) by Aspergillus nigerHA37 on a synthetic culture medium containing tannic acid at different concentrations has been studied. Maximal enzymatic activity increased according to the initial concentration of tannic acid; respectively 0.6, 0.9 and 1.5 enzyme activity units (EU) ml⁻¹ medium in the presence of 0.2%, 0.5% and 1% of tannic acid. Tannase production by A. niger HA37 on fourfold diluted olive mill waste waters (OMWW) as substrate, was between 0.37 and 0.65 EU ml⁻¹. Enzyme production on the diluted OMWW remained globally stable during more than 30 h. Growth of A. niger HA37 on OMWW was correlated with about 70% degradation of phenolic compounds present in the waste. This strain has therefore the capacity to degrade complex wastewaters which cause environmental damage to aquatic streams.     

Microbial solubilization of rock phosphate on media containing agro-industrial wastes and effect of the resulting products on plant growth and P uptake

Four agro-industrial wastes were assayed as substrates for microbial solubilization of rock phosphate (RP). Sugar beet wastes (SB), olive cake (OC) and olive mill wastewaters (OMWW) were treated by Aspergillus niger, and dry olive cake (DOC) was treated by Phanerochaete chrysosporium. In conditions of solid-state fermentation 46% of SB and 21% of OC were mineralized by A. niger while 16% of DOC was mineralized by P. chrysosporium. Repeated-batch mode of fermentation was employed for treatment of OMWW by immobilized A. niger, which resulted in conversion of 80% of the fermentable sugars. Acidification of all media treated by A. niger was registered with a simultaneous solubilization of 59.7% (SB), 42.6% (OC), and 36.4% (OMWW) of the total P present in the RP. The same mechanism of RP solubilization was observed in DOC-based medium inoculated with P. chrysosporium but other mechanisms were probably involved during the process. A series of microcosm experiments were then performed in the greenhouse to evaluate the effectiveness of the resulting fermented products. All amendments improved plant growth and P acquisition, which were further enhanced by mycorrhizal inoculation. The level of all studied parameters including the root mycorrhizal colonization depended on the substrate characteristics. The reported biotechnological schemes offer a potential application particularly for degraded soils.     

Olive mill waste as a substrate for nitrogen fixation

Olive mill wastewaters (OMWW) because of their low content in nitrogenous organic components and reachness in carbon sources offer a highly favourable environment for the growth of free-living dinitrogen fixing microorganisms. This property is manifested both in natural environments and in axenic cultures. Repetitive addition of OMWW to soil under aerobic conditions leads progressively to its enrichment with dinitrogen fixers, the activity of which is beneficial to soil fertility. The microbial consortium that develops in soil is dominated mostly by members of Azotobacter. A very efficient N₂-fixing and slime producing strain of Azotobacter vinelandii (strain A) was isolated from such an enriched soil sample. The isolate is deposited in the culture collection of our laboratory and its biochemical and molecular characteristics are investigated. The strain proved to be effective in bio-remediation processes of OMWW both in a laboratory-scale fermenter unit and a field pilot plant of ca 5 m³ capacity. The inhibitory growth-limiting components of the principal OMWW constituents and their impact on the duration of the lag period of N₂-fixing activity recovery is examined. The design of a multi-stream two stage process is described which provides a stable N₂-fixing system suitable for the bio-transformation of OMWW into an agrobiological product and/or for the production of extracellular polysaccharide ‘slime’ in high yields.     

Characteristics of the methane fermentation of waste of animal husbandry complexes with respect to the biomass of microorganisms and volatile fatty acids

The content of microbial biomass and the concentration of volatile fatty acids were comparatively studied in the tanks where the waste products of three stock farms were subjected to methane fermentation. The biomass content was shown to vary from 19 to 30% of the dry matter weight and the concentration of volatile fatty acids from 0 to 4.5 g per litre. The low concentrations of acetic and propionic acids together with the high biomass content were indicative of an active balanced microbial association. The parameters can be used to assess the operation of methane fermentation tanks.     

Production of <Emphasis Type="Italic">Pleurotus sajor-caju</Emphasis> strain PS-2001 biomass in submerged culture

Mushrooms or fruiting bodies of many basidiomycetes are commonly produced in solid-state fermentation, generally after 20-60 days of growth. However, it is also possible to produce biomass from these fungi, in submerged fermentation in shorter time. This work was aimed at evaluating biomass production with the basidiomycete Pleurotus sajor-caju, in a submerged process and to determine the proportion of chemical components of this biomass. Initially, an optimization of the culture medium was done to produce a faster growth of microbial mass by changing the concentrations of ammonium sulfate, soy protein and yeast extract. Using the optimized culture medium, values of approximately 5.5 g L(-1) of biomass in a medium with 10 g L(-1) of glucose were attained. When the optimized culture medium was tested in a 5-L stirred tank bioreactor, using 10 g L(-1) of glucose or sucrose as carbon source, values of 8.18 and 5.94 g L(-1) of biomass concentration were obtained, respectively. In the medium with glucose, high yields (0.82 g g(-1)) and productivity of 0.085 g L(-1) h(-1) were obtained. The exopolysaccharide content (1.58 g dry matter L(-1)) in the culture was higher in the fermentation with sucrose. The nutritional composition of the biomass obtained in the submerged fermentation was similar to that of the fruiting body in terms of quantities of total carbohydrates, ash and calories, but total fat and protein were higher.     

Anaerobic biotechnological approaches for production of liquid energy carriers from biomass

In recent years, increasing attention has been paid to the use of renewable biomass for energy production. Anaerobic biotechnological approaches for production of liquid energy carriers (ethanol and a mixture of acetone, butanol and ethanol) from biomass can be employed to decrease environmental pollution and reduce dependency on fossil fuels. There are two major biological processes that can convert biomass to liquid energy carriers via anaerobic biological breakdown of organic matter: ethanol fermentation and mixed acetone, butanol, ethanol (ABE) fermentation. The specific product formation is determined by substrates and microbial communities available as well as the operating conditions applied. In this review, we evaluate the recent biotechnological approaches employed in ethanol and ABE fermentation. Practical applicability of different technologies is discussed taking into account the microbiology and biochemistry of the processes.     

Influence of the flocculating agent in sedimentation and performance of a non flocculent strain of Zymomonas mobilis in the ethanol production process

The influence of the flocculating agent was studied in the performance (measured by microbial growth and ethanol production) of a non flocculent strain of Zymomonas mobilis, as well as the potentiality of the sedimentation process in the separation of the biomass from the fermentation broth. Among the flocculating agents studied, it was verified that both tannin and the polyelectrolyte yielded good results with regard to cellular performance. However, with regard to sedimentation tannin is more adequate to be used in processes involving Zymomonas mobilis.     

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.     

Inhibition of <Emphasis Type="Italic">Pichia stipitis</Emphasis> fermentation of hydrolysates from olive tree cuttings

The ethanolic fermentation of liquid fractions (hydrolysates) issued from dilute acid pre-treatment of olive tree biomass by Pichia stipitis is reported for the first time. On the one side, P. stipitis has been reported as the most promising naturally occurring C5 fermenting microorganism; on the other side, olive tree biomass is a renewable, low cost, and lacking of alternatives agricultural residue especially abundant in Mediterranean countries. The study was performed in two steps. First, the fermentation performance of P. stipitis was evaluated on a fermentation medium also containing the main inhibitors found in these hydrolysates (acetic acid, formic acid, and furfural), as well as glucose and xylose as carbon sources. The effect of inhibitors, individually or in a mixture, on kinetic and yield parameters was calculated. In a second step, hydrolysates obtained from 1% (w/w) sulfuric acid pre-treatment of olive tree biomass at 190°C for 10 min were used as a real fermentation medium with the same microorganism. Due to inhibition, effective fermentation required dilution of the hydrolysate and either overliming or activated charcoal treatment. Results show that ethanol yields obtained from hydrolysates, ranging from 0.35 to 0.42 g/g, are similar to those from synthetic medium, although the process proceeds at lower rates. Inhibiting compounds affect the fermentation performance in a synergistic way. Furfural is rapidly assimilated by the yeast; acetic acid and formic acid concentrations decrease slowly during the process. Activated charcoal or overliming detoxification improve the fermentability of diluted hydrolysates.     

Assessment of the genotoxicity of olive mill waste water (OMWW) with the Vicia faba micronucleus test

The present study concerns the genotoxicity of olive mill waste water (OMWW) generated in mills producing olive oil in Morocco. The Vicia faba micronucleus test was used to evaluate the genotoxicity of OMWW and the six major phenolic compounds identified by HPLC in this effluent. Five dilutions of OMWW were tested: 0.1, 1, 5, 10 and 20%. Maleic hydrazide was used as a positive control. The results showed that OMWW was genotoxic at 10% dilution. In order to investigate the components involved in this genotoxicity, the six major phenols present in this effluent, oleuropein, gallic acid, 4-hydroxyphenyl acetic acid, caffeic acid, paracoumaric acid and veratric acid, were studied at concentrations corresponding to the genotoxic concentration of the OMWW itself. Two phenols, gallic acid and oleuropein induced a significant increase in micronucleus frequency in Vicia faba; the four other phenols had no significant genotoxic effect. These results suggest that under the experimental conditions of our assay, OMWW genotoxicity was associated with gallic acid and oleuropein.     

Microbial biomass production from rice straw by fermentation with Penicillium janthinellum

Production of microbial biomass through fermentation of pretreated rice straw using Penicillium janthinellum (St-F-3B) is reported, with emphasis on the use of non-effluent generating pretreatment procedures. The fungus readily metabolized a rice straw substrate that had been subjected to alkali pretreatment by steaming at atmospheric pressure followed by the neutralization of the alkali. The crude protein content of the microbial biomass averaged 15–20%. The fermentation could be carried out in aerated-agitated fermenters using fertilizer grade nutrient salts to produce a biomass with 17.5% crude protein. When operated on a semicontinuous basis using 20% of the previous batch as inoculum, successive batches produced a biomass product with 12–19% crude protein content in 48 h. The problems of developing a technology for protein from agricultural residues with particular reference to developing countries are discussed.     

Effect of n-dodecane on Crypthecodinium cohnii fermentations and DHA production

The potential use of n-dodecane as an oxygen vector for enhancement of Crypthecodinium cohnii growth and docosahexaenoic acid (DHA) production was studied. The volumetric fraction of oxygen vector influenced the gas–liquid volumetric mass transfer coefficient k _L a positively. The k _L a increased almost linearly with the increase of volumetric fraction of n-dodecane up to 1%. The stirring rate showed a higher influence on the k _L a than the aeration rate. The effects of this hydrocarbon on C. cohnii growth and DHA production were then investigated. A control batch fermentation without n-dodecane addition (CF) and a batch fermentation where n-dodecane 1% (v/v) was added (DF) were carried out simultaneously under the same experimental conditions. It was found that, before 86.7 h of fermentation, the biomass concentration, the specific growth rate, the DHA, and total fatty acids (TFA) production were higher in the CF. After this fermentation time, the biomass concentration, the DHA and TFA production were higher in the DF. The highest DHA content of biomass (6.14%), DHA percentage of TFA (51%), and DHA production volumetric rate r _DHA (9.75 mg l⁻¹ h⁻¹) were obtained at the end of the fermentation with n-dodecane (135.2 h). The dissolved oxygen tension (DOT) was always higher in the DF, indicating a better oxygen transfer due to the oxygen vector presence. However, since the other C. cohnii unsaturated fatty acids percentages did not increase with the oxygen availability increase due to the n-dodecane presence, a desaturase oxygen-dependent mechanism involved in the C. cohnii DHA biosynthesis was not considered to explain the DHA production increase. A selective extraction through the n-dodecane was suggested.     

Kinetics of consumption of fermentation products by anode-respiring bacteria

We determined the kinetic response of a community of anode-respiring bacteria oxidizing a mixture of the most common fermentation products: acetate, butyrate, propionate, ethanol, and hydrogen. We acclimated the community by performing three consecutive batch experiments in a microbial electrolytic cell (MEC) containing a mixture of the fermentation products. During the consecutive-batch experiments, the coulombic efficiency and start-up period improved with each step. We used the acclimated biofilm to start continuous experiments in an MEC, in which we controlled the anode potential using a potentiostat. During the continuous experiments, we tested each individual substrate at a range of anode potentials and substrate concentrations. Our results show low current densities for butyrate and hydrogen, but high current densities for propionate, acetate, and ethanol (maximum values are 1.6, 9.0, and 8.2 A/m², respectively). Acetate showed a high coulombic efficiency (86%) compared to ethanol and propionate (49 and 41%, respectively). High methane concentrations inside the MEC during ethanol experiments suggest that methanogenesis is one reason why the coulombic efficiency was lower than that of acetate. Our results provide kinetic parameters, such as the anode overpotential, the maximum current density, and the Monod half-saturation constant, that are needed for model development when using a mixture of fermentation products. When we provided no electron donor, we measured current due to endogenous decay of biomass (~0.07 A/m²) and an open-cell potential (−0.54 V vs Ag/AgCl) associated with biomass components active in endogenous respiration. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Distribution of assimilated carbon in plants and rhizosphere soil of basket willow (Salix viminalis L.)

Willow is often used in bio-energy plantations for its potential to function as a renewable energy source, but knowledge about its effect on soil carbon dynamics is limited. Therefore, we investigated the temporal variation in carbon dynamics in willow, focusing on below-ground allocation and sequestration to soil carbon pools. Basket willow plants (Salix viminalis L.) in their second year of growth were grown in pots in a greenhouse. At five times during the plants growth, namely 0, 1, 2, 3 and 4 months after breaking winter dormancy, a subset of the plants were continuously labelled with ¹⁴CO₂ in an ESPAS growth chamber for 28 days. After the labelling, the plants were harvested and separated into leaves, first and second year stems and roots. The soil was analysed for total C and ¹⁴C content as well as soil microbial biomass. Immediately after breaking dormancy, carbon stored in the first year stems was relocated to developing roots and leaves. Almost half the newly assimilated C was used for leaf development the first month of growth, dropping to below 15% in the older plants. Within the second month of growth, secondary growth of the stem became the largest carbon sink in the system, and remained so for the older age classes. Between 31 and 41% of the recovered ¹⁴C was allocated to below-ground pools. While the fraction of assimilated ¹⁴C in roots and root+soil respiration did not vary with plant age, the amount allocated to soil and soil microbial biomass increased in the older plants, indicating an increasing rhizodeposition. The total amount of soil microbial biomass was 30% larger in the oldest age class than in an unplanted control soil. The results demonstrate a close linkage between photosynthesis and below-ground carbon dynamics. Up to 13% of the microbial biomass consisted of carbon assimilated by the willows within the past 4 weeks, up to 11% of the recovered ¹⁴C was found as soil organic matter.     

Sugar fermentation by <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> to produce ethanol

As a first step in the research on ethanol production from lignocellulose residues, sugar fermentation by Fusarium oxysporum in oxygen-limited conditions is studied in this work. As a substrate, solutions of arabinose, glucose, xylose and glucose/xylose mixtures are employed. The main kinetic and yield parameters of the process are determined according to a time-dependent model. The microorganism growth is characterized by the maximum specific growth rate and biomass productivity, the substrate consumption is studied through the specific consumption rate and biomass yield, and the product formation via the specific production rate and product yields. In conclusion, F. oxysporum can convert glucose and xylose into ethanol with product yields of 0.38 and 0.25, respectively; when using a glucose/xylose mixture as carbon source, the sugars are utilized sequentially and a maximum value of 0.28 g/g ethanol yield is determined from a 50% glucose/50% xylose mixture. Although fermentation performance by F.␣oxysporum is somewhat lower than that of other fermenting microorganisms, its ability for simultaneous lignocellulose-residue saccharification and fermentation is considered as a potential advantage.     

香菇液体发酵高产多糖菌株筛选

通过评价香菇野生菌株发酵产多糖性能,筛选高产香菇多糖菌株.以采自长白山野生香菇通过组织分离获得的6株菌株和2株人工栽培菌株为出发菌株,对不同发酵培养时间菌丝体生物量、胞内多糖含量、胞外多糖含量等进行测试分析,结果表明,8株菌株随着培养时间的延长,菌丝体生物量均有不同程度的增加,但胞内多糖含量和胞外多糖得率变化趋势不同,...     

Modeling of yeast metabolism and process dynamics in batch fermentation

Much is known about yeast metabolism and the kinetics of industrial batch fermentation processes. In this study, however, we provide the first tool to evaluate the dynamic interaction that exists between them. A stoichiometric model, using wine fermentation as a case study, was constructed to simulate batch cultures of Saccharomyces cerevisiae. Five differential equations describe the evolution of the main metabolites and biomass in the fermentation tank, while a set of underdetermined linear algebraic equations models the pseudo-steady-state microbial metabolism. Specific links between process variables and the reaction rates of metabolic pathways represent microorganism adaptation to environmental changes in the culture. Adaptation requirements to changes in the environment, optimal growth, and homeostasis were set as the physiological objectives. A linear programming routine was used to define optimal metabolic mass flux distribution at each instant throughout the process. The kinetics of the process arise from the dynamic interaction between the environment and metabolic flux distribution. The model assessed the effect of nitrogen starvation and ethanol toxicity in wine fermentation and it was able to simulate fermentation profiles qualitatively, while experimental fermentation yields were reproduced successfully as well.