Microbiological Production of Gibberellic Acid in Glucose Media

Gibberellic acid production from various substrates was studied in 43 strains of Fusarium, among which F. moniliforme strain IOC-3326 was selected as the best producer. Experiments were carried out in shaker flasks and pilot plant fermentors. The results indicate that the best substrate for gibberellic acid production with this strain is composed of the following: glucose, 20 g; corn steep liquor, 25 g; ammonium nitrate, 2.6 g; monopotassium phosphate, 0.5 g; potassium sulfate, 0.2 g; and water, 1000 ml. Glucose, ammonium nitrate, and corn steep liquor were found to be critical. With this medium, maximal yields of 1196 mg per liter in shaker flasks and 997 mg per liter in fermentors were produced.     

Quantitative estimation of gibberellic acid by paper chromatography

Метод для количественного определения от gibberellic кислоты в процессе брожения средства массовой информации, с использованием бумаги по убыванию хроматографии в butylacetate воды описана. Образца корректируется, чтобы рН 2.5-3.0, добыто с н-бутанола, и 0,05 мл. органического слоя пятнами на Хроматографический бумагу. После equilibration от Атмосфера в банке, chromatogram Разработана в butylacetate насыщенных с водой, за 7 часов, и растворитель разрешено покинуть капельного нижней части листа. Обнаружение осуществляется путем опрыскивания с 3% раствор серной кислоты в метаноле и после сушки бумаги, пятна с синий u.v. флуоресценции наблюдается. Определенный артикль площадь пятна оценивается с помощью калибровочной кривой, заговор с ценностей, стандартов, соответствующих 20, 60 и 120 μ g. gibberellic кислоты. Погрешность оценки составляет ± 10-15% когда оценки выполняются тщательно. Низкий предел чувствительности 5 μg.     

Paecilomyces sp. ZB is a cell factory for the production of gibberellic acid using a cheap substrate in solid state fermentation

Gibberellic acid from the fungi has been widely used in agriculture. In this study, more than 20 fungal isolates were screened and Paecilomyces sp. ZB shown to produce more gibberellic acid than other fungal isolates. Cow dung was used as low cost substrate for gibberellic acid production in solid state fermentation (SSF). Carbon, nitrogen and ionic sources stimulated gibberellic acid production in SSF. Lactose emerged as the significant carbon source supporting more gibberellic acid production (731 µg/g). Among the nitrogen sources, glycine appeared to influence the production of more gibberellic acid (803 µg/g). The process parameters were optimized to enhance gibberellic acid production using a two-level full factorial design and response surface methodology. The amount of gibberellic acid production was influenced mainly by moisture and pH of the substrate. Gibberellic acid production was 1312 µg/g under the optimized conditions and the predicted response was 1339 µg/g. The gibberellic acid yield increased twofolds after medium optimization. The extracted gibberellic acid was sprayed on the growing Mung bean plant and it stimulated the growth of the plant effectively. To conclude, cow dung is a new alternative to produce gibberellic acid in SSF.     

Effects of cultivation conditions on the production of natamycin with Streptomyces gilvosporeus LK-196

Natamycin is a very attractive antifungal agent with wide applications in medical and food industries. In order to improve the productivity of natamycin, the effects of cultivation conditions were investigated with Streptomyces gilvosporeus LK-196 in the shake flasks and 30-L fermentors. The results showed that dissolved oxygen and shear force would affluence the biosynthesis of natamycin significantly. The high concentration of natamycin (2.03g/L) was achieved under the suitable culture conditions in the shake flask scale. Further investigations in 30-L fermentors showed that the optimal pH was controlled at 6.0 during the whole bioprocess, and the dissolved oxygen level should be more than 30% by adjusting the aeration and agitation rates for high production of natamycin. Under these optimal conditions the high concentration of natamycin (3.94g/L) was achieved with Str. gilvosporeus LK-196 in the 30-L fermentor. Finally, the high-level fermentation process was successfully scaled up to 1000-L fermentors and 18,000-L fermentors in the pilot plant.     

Fixed-bed fermentation of rice straw and chicken manure using a mixed culture of marine mesophilic microorganisms

A mixture of rice straw (80%) and chicken manure (20%) was pretreated and fermented to carboxylic acids by using a mixed culture of marine mesophilic microorganisms. Two sets of four fermentors, built from PVC pipes, were used for both biomass pretreatment and fermentation. Four 1L fermentors (F1-F4) were arranged in series, where liquid fermentation products were transferred from one fermentor to the other, to form a train. A liquid volume of 10mL and 15mL were transferred every four days for Trains A and B, respectively. The maximum total acid concentration for F1 in Train A was 34.2g/L and the maximum acid concentration in F2-F4 was approximately 44g/L. The maximum total acid concentration in F1 in Train B was 30.5g/L and the maximum acid concentration in F2-F4 was approximately 48g/L. The conversion in each of the fermentors in Train A varied from 0.821 to 0.879g VS digested/g VS fed and the yield was in the range 0.489-0.609g total acids/g VS fed. The conversion and yield in Train B were 0.741-0.914g VS digested/g VS fed and 0.563-0.669g total acids/g VS fed, respectively. The continuum particle distribution model (CPDM) predicted acid concentrations and retention times in the fixed-bed fermentation system with R(2) of 0.67-0.84 in Trains A and B.     

Effects of Benzo(a)pyrene on Seabass (Dicentrarchus labrax L.): Biomarkers,Growth and Behavior

The main purpose of this study was to investigate the effects of benzo(a)pyrene (BaP) on seabass (Dicentrarchus labrax) juveniles using parameters at different levels of biological organization. Liver antioxidant status, BaP biotransformation and accumulation, growth, and behavior were determined in juveniles after 28 d exposure to BaP (1–16 μ g/l). Liver ethoxyresorufin O-deethylase increased in seabass exposed to 1–8 μ g/l of BaP. Liver glutathione S-transferases and catalase activities were significantly increased at 4 and 8 μ g/l, but a slight decrease was observed at the highest concentrations tested. Bile BaP metabolites were significantly different from the control group at 1 and 16 μ g/l BaP. Liver BaP metabolites and lipid peroxidation significantly increased at 8 and 16 μ g/l BaP. These results suggest that BaP metabolites' accumulation induces oxidative damage in seabass liver. Body weight and length increase were significantly reduced in fish exposed to BaP, with LOECs of 16 and 4 μ g/l, respectively. Food intake and swimming velocity were significantly decreased after exposure to BaP, with LOEC values of 16 and 8 μ g/l, respectively. Results suggest that at concentrations of BaP equal or higher than 8 μ g/l, the detoxification capacity decreases, an accumulation of liver BaP metabolites occurs causing lipid peroxidation, affecting growth and swimming capability of fish.     

Continuous 2-Keto-l-gulonic acid fermentation by mixed culture of Ketogulonicigenium vulgare DSM 4025 and Bacillus megaterium or Xanthomonas maltophilia

The fermentation process of 2-keto-L-gulonic acid (2KGA) from L-sorbose was developed using a two-stage continuous fermentation system. The mixed culture of Ketogulonicigenium vulgare DSM 4025 and Bacillus megaterium DSM 4026 produced 90 g/L of 2KGA from 120 g/L of L-sorbose at the dilution rate of 0.01 h⁻¹ in a single-stage continuous fermentation process. But after the production period was beyond 150 h, the significant decrease of 2KGA productivity was observed. When the non-spore forming bacteria Xanthomonas maltophilia IFO 12692 was used instead of B. megaterium DSM 4026 as a partner strain for K. vulgare DSM 4025, the 2KGA productivity was significantly improved in a two-stage continuous culture mode, in which two fermentors of the same size and volume were connected in series. In this mode, with two sets of 3-L jar fermentors, the steady state could be continued to over 1,331.5 h at least, when the dilution rates were 0.0382 h⁻¹ and 0.0380 hour⁻¹, respectively, for the first and second fermentors. The overall productivity was calculated to be 2.15 g/L/h at 113.1 g/L and a molar conversion yield of 90.1%. In the up-scaling fermentation to 30-L jar fermentors, 118.5 g/L of 2KGA was produced when dilution rates in both stages were 0.0430 hour⁻¹, and the overall productivity was calculated to be 2.55 g/L/h.     

High-productivity fermentation process for cultivating industrial microorganisms

Summary High-productivity continuous fermentation processes have been developed for the production of important industrial microorganisms in specially designed fermentors.Saccharomyces cerevisiae, Pichia pastoris, Kluyveromyces fragilis, andCandida utilis yeasts have been grown in bench-scale fermentors at cell densities of over 120 g/l, whileEscherichia coli, Bacillus megaterium, Methylomonas sp. andPseudomonas putida bacteria have been cultivated to cell densities of more than 110 g/l. Productivities (g cells per 1 per h) greater than 25 have been achieved in both bench-scale and 1500-liter fermentors with yeasts, and values as high as 55 have been achieved with bacteria in the bench-scale fermentor. The microorganisms were grown on defined media using ammonia for pH control and as nitrogen source. The fermentor, capable of high oxygen and heat transfer rates, was operated at constant volume with continuous feed and product discharge. The high-productivity process reduces fermentor size, media sterilization requirements, and may under some circumstances eliminate waste and recycle streams. It can also be applied to a variety of biological products.     

Solid-state fermentation systems-an overview

Starting with a brief history of solid-state fermentation (SSF), major aspects of SSF are reviewed, which include factors affecting SSF, biomass, fermentors, modeling, industrial microbial enzymes, organic acids, secondary metabolites, and bioremediation. Physico-chemical and environmental factors such as inoculum type, moisture and water activity, pH, temperature, substrate, particle size, aeration and agitation, nutritional factors, and oxygen and carbon dioxide affecting SSF are reviewed. The advantages of SSF over Submerged Fermentation (SmF) are indicated, and the different types of fermentors used in SSF described. The economic feasibilities of adopting SSF technology in the commercial production of industrial enzymes such as amylases, cellulases, xylanase, proteases, phytases, lipases, etc., organic acids such as citric acid and lactic acid, and secondary metabolites such as gibberellic acid, ergot alkaloids, and antibiotics such as penicillin, cyclosporin, cephamycin and tetracyclines are highlighted. The relevance of applying SSF technology in the production of mycotoxins, biofuels, and biocontrol agents is discussed, and the need for adopting SSF technology in bioremediation of toxic compounds, biological detoxication of agro-industrial residues, and biotransformation of agro-products and residues is emphasized.     

Solid-State Fermentation Systems—An Overview

Abstract

Starting with a brief history of solid-state fermentation (SSF), major aspects of SSF are reviewed, which include factors affecting SSF, biomass, fermentors, modeling, industrial microbial enzymes, organic acids, secondary metabolites, and bioremediation. Physico-chemical and environmental factors such as inoculum type, moisture and water activity, pH, temperature, substrate, particle size, aeration and agitation, nutritional factors, and oxygen and carbon dioxide affecting SSF are reviewed. The advantages of SSF over Submerged Fermentation (SmF) are indicated, and the different types of fermentors used in SSF described. The economic feasibilities of adopting SSF technology in the commercial production of industrial enzymes such as amylases, cellulases, xylanase, proteases, phytases, lipases, etc., organic acids such as citric acid and lactic acid, and secondary metabolites such as gibberellic acid, ergot alkaloids, and antibiotics such as penicillin, cyclosporin, cephamycin and tetracyclines are highlighted. The relevance of applying SSF technology in the production of mycotoxins, biofuels, and biocontrol agents is discussed, and the need for adopting SSF technology in bioremediation of toxic compounds, biological detoxication of agro-industrial residues, and biotransformation of agro-products and residues is emphasized.     

Biosurfactant yields and nutrient consumption of Pseudomonas fluorescens 378 studied in a microcomputer controlled multifermentation system

Production of biosurfactant AP-6 and consumption of carbon (succinic acid) and nitrogen (ammonium ions) by Pseudomonas fluorescens 378 were studied under different growth conditions. The study was performed in a microcomputer controlled multibatch fermentation system which enabled simultaneous running of 10 fermentors. The fermentors were mantled glass vessels, temperature controlled by circulated water, and mixing was arranged by magnetic stirrers. They were connected to the computer system (pH measurement and control) via signal conditioning cards. The microcomputer had a 128 kbytes RAM, two 800-kbyte floppy disc drives, a graphic terminal, and expansion cards. Biosurfactant production was independent of the carbon-to-nitrogen ratio and the phosphorus content in the medium. Omitting the Fe(III) supplement to the medium increased the product yield by 120%. Changes in oxygen transfer rate and pH in the iron deficient cultures did not have any effect on the product yield. Iron deficiency increased the cell consumption of carbon source. Consumption of carbon source in relation to nitrogen uptake (carbon/nitrogen quotient) increased with increasing quotient in the growth medium. The uptake of carbon and nitrogen changed in the intervals of 1.2-1.5 g/g biomass and 0.09-0.16 g/g biomass, respectively. The consumption of carbon increased from 1.5 g/g biomass to 2.0 g/g biomass when the medium concentration of phosphorus was decreased from 0.18 to 0.027 g/L.     

Synergism between Gibberellin and Auxin in the Growth of Intact Tomato

Gibberellin A_4&7 was more effective than gibberellic acid in increasing shoot elongation when applied to the apex of intact Lycopersicum esculentum seedlings of Tiny Tim, a dwarf cultivar, and Winsall, a tall cultivar. After 14 days, gibberellic acid and gibberellin A_4&7 stimulated growth of the dwarf more than the tall tomato. In tall tomato the application of indole-3-acetic acid alone (6.1 μg/plant) showed an inhibitory growth effect, but when applied with 17.5 μg per plant of gibberellic acid, it had a synergistic effect at 7 days but not at 14 days. When the auxin concentration was reduced to 0.61 μg per plant a synergistic effect was observed on tall plants at 7 and 14 days between indole-3-acetic acid and gibberellic acid. Application of gibberellin A_4&7 with auxin did not give a synergistic response in tall or dwarf tomato.     

Effect of 4-methyl-2-thiouracil on growth of leuconostoc mesenteroides

Эффект goitrogen 4-метил-2-Thiouracil конц е нтраций50—2,000 μg./10 ml.средни й п о росту L. mesenteroidesизучала сь. Это вещество, торм озящ ий эффект по рос ту L. mesenteroides то лько в средст ва массово й информации, содерж ащи е пиридоксин. Ее эффект связана с кон центрацие й и Присутствие пи ридо ксин. Высокая концентр ация пири доксин(5.0 μg./10 ml. medium)з аметно снижает эфф ект даже 2.000 μ g. 4-methyl-2-thiouracil/10 мл сре дой И нгибирование ростаL. mesenteroides 4-метил-2-thiouracil упр аздняется конкурсной основе thymine пу тем, а не на конкур сной п утем thymidine и folie кисло ты до кон центрации 1.000 μ g. 4-methyl-2-thiouracil/10 мл. ср еднесрочной.     

Continuous fermentation to produce fungal protein. Effect of growth rate on the biomass yield and chemical composition of Fusarium moniliforme

Fusarium moniliforme was grown on a carob aqueous extract in a chemostat for fungal protein production. The substrate was adjusted to provide 0.5% carob sugars supplemented with inorganic salts. The dilution rate varied from 0.086 to 0.227 hr^?1 under constant conditions of temperature (30°C), pH (4.5), and oxygen saturation (60–80%). A yield of 0.709 g dry mycelium/g consumed carob sugar and a productivity value of 0.687 g dry mycelium/liter hr^?1 were obtained at μ = 0.205 hr^?1. The maintenance coefficient was 0.077 g carob sugar/g dry mycelium hr^?1. While the carbohydrate and purine content of dry mycelium increased at μ values from 0.114 to 0.205 hr^?1 both true (Lowry) and crude (N × 6.25) protein contents decreased at the same μ range. Maximum values of 36.3% true and 47.9% crude protein of dry mycelium were obtained at μ = 0.114 hr^?1, whereas a minimum purine content of 99.8 μmol/g corresponding to 6.42% nucleic acids was recorded at μ = 0.086 hr^?1. It was concluded that a continuous fermentation of carob aqueous extract using F. moniliforme should be operated at growth rates of approximately 0.205 hr^?1 in order to maximize protein production.     

自絮凝酵母SPSC01在组合反应器系统中酒精连续发酵的研究

建立了一套由四级磁力搅拌发酵罐串联组成、总有效容积4000mL的小型组合生物反应器系统 ,其中一级罐作为种子培养罐。以脱胚脱皮玉米粉双酶法制备的糖化液为种子培养基和发酵底物 ,进行了自絮凝颗粒酵母酒精连续发酵的研究。种子罐培养基还原糖浓度为100g L ,添加 (NH₄)₂HPO₄ 和KH₂PO₄ 各 20g L ,以0.017h^-1 的恒定稀释速率流加 ,并溢流至后续酒精发酵系统。发酵底物初始还原糖浓度 220g/L ,添加 (NH₄)₂HPO₄ 15g/L和KH₂PO₄2 5g/L ,流加至第一级发酵罐 ,稀释速率分别为 0.017、0.025、0.033、0.040和0.05 0h^-1。实验数据表明 ,自絮凝颗粒酵母在各发酵罐中呈部分固定化状态 ,在稀释速率0.040h^-1 条件下 ,发酵系统呈一定的振荡行为 ,其他四个稀释速率实验组均能够达拟稳态。当稀释速率不超过 0 0 33h^-1 ,流出末级发酵罐的发酵液中酒精浓度可以达到 12 % (V/V)以上 ,残还原糖和残总糖分别在 0 11%和 0 35 % h^-1,流出末级发酵罐的发酵液中酒精浓度可以达到12%（V/V）以上,残还原糖和残总糖分别在0.11%和0.35%（W/V）以下。在稀释速率为0.033h^-1时,计算发酵系统酒精的设备生产强度指标为3.32（g·L^-1·h^-1）,与游离酵母细胞传统酒精发酵工艺相比,增加约1倍。     

Methane Production by Fermentor Cultures Acclimated to Waste from Cattle Fed Monensin, Lasalocid, Salinomycin, or Avoparcin

The ability of microorganisms to ferment waste from cattle fed monensin, lasalocid, or salinomycin to methane was determined. Continuously mixed anaerobic fermentors with 3-liter working volumes at 55°C were used; fermentors were fed once per day. Initially, all fermentors were fed waste without antibiotics at 6% volatile solids (VSs, organic matter) and a 20-day retention time (RT) for 60 days. Waste from animals fed monensin, lasalocid, or salinomycin at 29, 20, and 16.5 mg per kg of feed, respectively, was added to duplicate fermentors at the above VSs, and RT. Avoparcin (5 to 45 mg/liter) was not fed to animals but was added directly to duplicate fermentors. Lasalocid and salinomycin had minimal effects on the rate of methane production at RTs of 20 days and later at 6.5 days. Avoparcin caused an increase in organic acids from 599 to 1,672 mg/liter (as acetate) after 4 weeks, but by 6 weeks, acid concentrations declined and the rate of methane production was similar to controls at a 6.5-day RT. The monensin fermentors stopped producing methane 3 weeks after antibiotic addition. However, after a 6-month acclimation period, the microorganisms apparently adapted, and methane production rates of 1.65 and 2.51 liters per liter of fermentor volume per day were obtained with 6% VSs, and RTs of 10 and 6.5 days, respectively. This compares with 1.78 and 2.62 liters/liter per day for controls (P > 0.05). All fermentors that were fed waste containing antibiotics had lower pH values and ammonia and alkalinity concentrations, suggesting less buffering capacity and protein catabolism than in controls. Acclimation results obtained with fermentors at 35°C were similar to those for fermentors at 55°C. These studies indicate that waste from cattle fed these selected growth-promoting antibiotics can be thermophilically fermented to methane at RTs of 6.5 days or longer and VS concentrations of 6%, at rates comparable to waste without antibiotics.     

Influence of Matrix Composition on the Bioaccessibility of Copper,Zinc, and Nickel in Urban Residential Dust and Soil

This study examines factors affecting oral bioaccessibility of metals in household dust, in particular metal speciation, organic carbon content, and particle size, with the goal of addressing risk assessment information requirements. Investigation of copper (Cu) and zinc (Zn) speciation in two size fractions of dust (< 36 μ m and 80–150 μ m) using synchrotron X-ray absorption spectroscopy (XAS) indicates that the two metals are bound to different components of the dust: Cu is predominately associated with the organic phase of the dust, while Zn is predominately associated with the mineral fraction. Total and bioaccessible Cu, nickel (Ni), and Zn were determined (on dry weight basis) in the < 150 μ m size fraction of a set of archived indoor dust samples (n = 63) and corresponding garden soil samples (n = 66) from the City of Ottawa, Canada. The median bioaccessible Cu content is 66 μ g g^?1 in dust compared to 5 μ g g^?1 in soil; the median bioaccessible Ni content is 16 μ g g^?1 in dust compared to 2 μ g g^?1 in soil; and the median bioaccessible Zn content is 410 μ g g^?1 in dust compared to 18 μ g g^?1 in soil. For the same data set, the median total Cu content is 152 μ g g^?1 in dust compared to 17 μ g g^?1 in soil; the median total Ni content is 41 μ g g^?1 in dust compared to 13 μ g g^?1 in soil; and the median total Zn content is 626 μ g g^?1 in dust compared to 84 μ g g^?1 in soil. Organic carbon is elevated in indoor dust (median 28%) compared to soil (median 5%), and is a key factor controlling metal partitioning and therefore bioaccessibility. The results show that house dust and soil have distinct geochemical signatures and should not be treated as identical media in exposure and risk assessments. Separate measurements of the indoor and outdoor environment are essential to improve the accuracy of residential risk assessments.     

Efficient acetic acid production by repeated fed-batch fermentation using two fermentors

Summary In acetic acid fermentation, the number of viable cells decrease as the acetic acid concentration increases to more than about 40 g/l, which means that the productivity attainable by conventional fed-batch and repeated fed-batch operations using one fermentor is limited. In this paper, based on a fed-batch experiment using Acetobacter aceti 2096, a mathematical model was developed. The optimization carried out showed the superiority of repeated fed-batch operation using two fermentors. The performance evaluation was made with respect to productivity and product concentration. It was shown to be attractive in practice to use multiple fermentors, in particular for high product concentrations. Experiments were then conducted to ascertain the simulation results. Offprint requests to: T. Kobayashi     

Apparatus for monitoring the oxygen uptake and carbon dioxide production of fermentations

The requirements of the continuous analysis of effluent gas streams from aerated flash and tank fermentors are described, as are instrumental devices for measuring the oxygen and carbon dioxide content of fermentor gases. The use of a specially designed sequential gas sample for monitoring four fermentations simultaneously and a system for precise control of low air flow and pressure is explained. Equations for calculating carbon dioxide production or oxygen consumption rates and respiratory quotients are given. A discussion of the operating characteristics of a device for automatic translation of aeration data between fermentors is presented.     

Spectrophotometric method for determining gibberellic acid in fermentation broths

A novel method for the quantitative determination of gibberellic acid in fermentation broths has been developed. It is based on the kinetic of the reaction of conversion of gibberellic acid to gibberellenic acid. The method is simple, reliable, faster than most of methods known, and free of the interferences which commonly affect spectrophotometric methods currently in use. Its threshold sensitivity is 0.1 g and its accuracy is greater than 97% for concentrations of gibberellic acid ranging from 0.1 to 1 g l(-1).