Optimization of growth ofEscherichia alcalescens with respect to its aspartate ammonia-lyase activity

Aspartate ammonia-lyase activity ofEscherichia alcalescens was positively affected by the composition of the culture medium and by a higher intensity of aeration. By using the supernatant obtained from autolyzed baker’s yeast, the aspartate ammonia-lyase activity increased two-fold (about 90 μkat/g wet biomass) as compared to commercial yeast autolyzates. The authors thank Mr. J. Tomíček (Food Industry Research Institute, Prague) for a sample of the yeast autolyzate of the Kolín type.     

Effect of boron application on barley in a sierozem sandy soil

Summary In a pot culture experiment on a sierozem sandy soil (pH 8.2) rates of added B at 3 ppm although decreased root yield significantly but shoot and grain yield was unaffected even at 6 ppm added B, even though shoot B concentration was as high as 360 ppm and Ca/B ratio as low as 11. At 6 ppm applied B, shoot yield was increased by 18.5 per cent, whereas grain yield was at par with control. The results suggested that Ca/B ratio in barley straw was not a reliable index for determing the magnitude of B problem in the soil.     

Two bacteria causing farmer's lung: Fine structure ofThermoactinomyces vulgaris andSaccharopolyspora rectivirgula

Larval cuticle ofHelicoverpa (Heliothis)zea and yeast extract added to a minimal medium (MM) induced germination of conidia ofNomuraea rileyi whereas sterile distilled water or MM alone did not. Yeast extract increased mycelial yield, but when cuticle was added, mycelial yield significantly decreased. Proteases and chitinases ofN. rileyi were only expressed when cuticle was added to the MM.This article reports the results of research only. Mention of a proprietary product in this paper does not constitute a recommendation for use by US Department of Agriculture.     

Sporogenicity of yeast autolyzates and casein hydrolyzates for Bacillus popilliae in liquid cultures

Bacillus popilliae NRRL B-2309S forms several million refractile spores/ml in liquid shaken cultures. A suitable medium includes glucose, K₂HPO₄, water, and three selected ingredients-activated carbon, a yeast autolyzate, and a casein hydrolyzate. Only 4 out of 26 lots of commercial yeast autolyzates tried were sporogenic. However, spore formation in the presence of the four was poor and erratic unless a compatible casein hydrolyzate also was present. Five of eight lots of casein hydrolyzates improved the sporogenicity of selected yeast products to various degrees. A comparison of amino acid compositions of yeast and casein lysates sheds no light on differences between suitable and unsuitable ones. Less refined yeast and casein products seem preferable.     

Effect of C/N ratio and starch concentration on ethanol production from sago starch using recombinant yeast

Recombinant Saccharomyces cerevisiae YKU 131 (capable of expressing glucoamylase) was used to produce ethanol from sago starch. The optimum C/N ratio for ethanol production by the recombinant yeast was 7.9, where 4.7 and 10.1 g/l ethanol was produced from 20 and 40 g/l sago starch, respectively. At sago starch concentration higher than 40 g/l and C/N ratio higher than 10.4, glucoamylase production and rate of starch hydrolysis were reduced, which in turn, reduced ethanol production significantly. The theoretical yield of ethanol based on sago starch consumed in fermentation using 40 g/l was 72.6%. This yield was slightly lower than those obtained in fermentation using soluble starch such as potato and corn starch, which ranged from 80–90% as reported in the literature. However, S. cerevisiae YKU 131 could only utilize 62% of the total amount of starch added to a medium.     

The influence of heavy metals upon the growth of sitka-spruce in South Wales forests

Summary Experiments on sitka-spruce seedlings grown in acidic peaty gley soils under green-house conditions, where the soils where doped with increasing amounts of Cd, Cu and Pb up to maximum levels of metal added of 16 ppm, 32 ppm and 400 ppm respectively, showed that the levels of Cd and Pb in shoots and roots increased with increasing levels in the soil, whereas levels of copper appeared to be independent. The addition of these three metals to the soils did not influence the uptake of other heavy metals, or of the nutrients potassium or calcium. Increases in the shoot cadmium levels significantly reduced the yields of the plant shoots. However, the plant yields were only affected by the highest level of lead that was added to the soil (400 ppm Pb) and unaffected by all the copper treatments (0–32 ppm Cu in the soil). The lengths of the sitka-spruce roots were reduced when cadmium and lead levels in the soil exceeded certain threshold concentrations (2.5 ppm total Cd, where 0.3 ppm was extractable with 0.5 M acetic acid; and 48 ppm total Pb, where 1.7 ppm was extractable). However, root lengths were not reduced by copper. This was probably related to the fact that copper appears to be relatively unavailable in the type of soil used, as only 1.1. ppm Cu was extractable from a total of 32 ppm Cu added. Root branching was apparently reduced by increases in the soil levels of cadmium, copper and lead. The roots of some control plants had symbiotic mycorrhizal associations (4 out of 19 plants), whereas the roots of all the plants grown in the soils with added heavy metals did not develop these.     

Comparative study of the effect of ferrocyanide and EDTA on the production of ethyl alcohol from molasses by Saccharomyces cerevisiae

The effects of potassium ferrocyanide and EDTA on ethyl alcohol production from molasses by Saccharomyces cerevisiae were investigated on simulated batch pilotplant-scale conditions for alcoholic fermentation of molasses. Ethyl alcohol production was more sensitive to ferrocyanide than to EDTA. When ferrocyanide was introduced into the cultures at the time of inoculation, there was stimulation of ethyl alcohol production, with 261 ppm ferrocyanide producing the maximum effect, which was 3.0% more than in control cultures. When added during the propagation of the yeast, ferrocyanide depressed ethyl alcohol production by 4.0% maximum whereas EDTA stimulated ethyl alcohol production by 2.0%. Addition of ferrocyanide during the fermentation stage produced no significant effect on alcohol production, whereas over a wide range of EDTA concentration there was a steady increase in alcohol yield.     

Effect of irrigation treatments upon the chemical constituents ofHibiscus sabdariffa L.

Summary The effects of Se and P on the dry matter yield and chemical composition of Raya (Brassica juncea Cos. var. R.L. 18) were studied in the green-house. The dry matter yield in P treated and untreated pots increased with Se application upto 2.5 ppm. With further increase in Se dose upto 10 ppm, dry matter yield decreased. The increase in P dose from 0 to 50 ppm increased dry matter yield in all Se treated and untreated pots but 100 ppm P rather showed decrease in dry matter yield as compared to 50 ppm P.The Se concentration increased by about 100 fold with the application of 10 ppm Se over 0 ppm Se where P was not added whereas increase was 258 and 336 times when 50 ppm and 100 ppm P was added, respectively. Inorganic, organic and total P increased with increasing Se and P. The increase in inorganic P was more than organic and total P. S and N concentration decreased with increasing Se application and increased with P application.Crude protein showed the same behaviour as N. The total sulphur-containing amino acids (which ranged from 39 to 49% of crude protein) and individual sulphur-containing amino acids like methionine (16.9 tot 20%) cysteine (9.8 to 13%) and cystine (12.3 to 15.9%) decreased with the increasing selenium but increased with increasing phosphorus. N/S ratio in the plant showed significant negative correlation with total sulphur-containing amino acids (r=–0.940^**), methionine (r=–0.951^**), cysteine (r=–0.929^**) and cystine (r=–0.920^*) whereas total sulphur in the plant showed positive significant correlation with methionine (r=0.805^**), cysteine (r=0.924^**) and cystine (r=0.821^**).     

Influence of plant growth regulators on growth, morpho-physiological characters and yield of summer sesame (Sesamum indicum L. cv. Rama) under moisture stress

Field experiments were conducted with sesame (Sesamum indicum L. cv. Rama) for two years (1997 and 1998) to study the effect of three level of irrigation (F+C, B+C, B+F+C) and two growth regulators (CCC, 200 ppm CCC; 100 ppm and BX-112, 100 ppm; BX-112, 50 ppm) on growth (root and shoot length, average number of primary branches/plant), morpho-physiological growth parameters(LAI, LAD, CGR and NAR), yield attributing parameters(average number of capsule/plant, average number of seeds/capsule) and seed yield. Irrigation at B+F+C stage showed significant effect on these parameters. Among the growth regulators, CCC, 200 ppm showed remarkable results on these parameters and seed yield. Seed yield in CCC, 200 ppm treatment was more than 53% in comparison to water soaked seeds. The interaction between irrigation and PGR showed better seed yield and it was concluded that the growth regulator CCC might be utilized for enhancement of seed yield of summer sesame under field condition.     

Effects of addition of varying levels of copper,as oxide or phosphate,on nitrogen mineralisation and nitrification during incubation of a slightly calcareous soil receiving dried blood

Summary The addition of 100, 1000 and 10,000 ppm copper, as oxide or hydrogen phosphate, stimulated nitrogen mineralisation and nitrification during incubation of a sandy loam (0.5% calcium carbonate) treated with 200 ppm N as dried blood. The maximum effects occurred with 1000 ppm added copper and were similar with both sources of added copper. EDTA-extractable copper was higher where copper hydrogen phosphate than where copper oxide had been added.     

Effect of technical threonine sources on homoserine biosynthesis by mutant Brevibacteruim flavum 2T

The effect of threonine technical sources on the homoserine biosynthesis by the threonine auxotroph Brevibacterium flavum 2T when cultivated on sucrose and acetic acid containing media was investigated. Various threonine sources (corn extract and fodder yeast, microbial biomass and soybean meal hydrolyzates) prepared by means of different hydrolyzing agents (acids, enzymes, autolysis) were used. The most effective substrate was protein--vitamin concentrate hydrolyzate, particularly combined with corn extract in the ratio 1: 0,25-0.5 (with respect to the dry weight of the initial material). The homoserine yield was 16.2 g/l on the sucrose containing medium and 18.4 g/l on the acetic acid containing medium which was in agreement with controls. The medium containing pure threonine was used as a control. With other threonine sources (corn extract, protein-vitamin concentrate autolyzate and enzymolyzate, fodder yeast and soybean meal hydrolyzates), the homoserine production was significantly lower, i.e. 40-70% of the control. The content of amino acids (threonine, isoleucine, methionine) in the initial material and their suitability for the homoserine biosynthesis were found to be correlated. The substrates with a high content of threonine (over 3.5%) and a low content of methionine (below 0.5%) proved most effective. The use of the material in which the ratio threonine: methionine was less than 6.0 caused the homoserine biosynthesis to be partially replaced with that of lysine.     

Improving the ethanol yield by reducing glycerol formation using cofactor regulation in Saccharomyces cerevisiae

To increase ethanol yield and decrease glycerol production in Saccharomyces cerevisiae, the strategies of direct cofactor-regulation were explored. During anaerobic batch fermentations, the yeast expressing Bacillus cereus gapN gene, encoding non-phosphorylating NADP(+)-dependent glyceraldehyde-3-phosphate dehydrognease, produced 73.8?g ethanol?l(-1), corresponding to 96% of theoretical maximum yield compared to 92% for the wild type. The yeast expressing Escherichia coli frdA gene encoding the NAD(+)-dependent fumarate reductase, exhibited a 22% (relative to the amount of substrate consumed) increase in glycerol yield in medium containing 2?g fumarate?l(-1). The yeast expressing mhpF gene, encoding acetylating NAD(+)-dependent acetaldehyde dehydrogenase, produced 74.5?g ethanol?l(-1), corresponding to 97.4% of theoretical maximum yield while glycerol decreased by 40% when acetic acid was added before inoculation. This strain represents a promising alternative for ethanol production with lignocellulosic hydrolysates where acetate is available at significant amounts.     

Growth ofAureobasidium pullulans on waste water hemicelluloses

Strain Aureobasidium pullulans capable of utilizing hemicelluloses and xylan was cultivated on processed waste dialysis liquor from the production of viscose fibres, containing about 1.5% hemocelluloses. Basic conditions of biomass production were tested on a laboratory scale. The dialysis waste liquor adjusted with mineral acids to pH 4--5 and supplemented with 0.05% yeast autolyzate and 0.2% ammonium sulphate affords protein yields of about 0.8 g/l, corresponding to 4.0--4.5 g dry biomass. Biomass is isolated together with residual water-insoluble hemicelluloses which are not utilized by the microorganism. The total utilization of hemicelluloses attains about 70%.     

Formation by Yeast of the HEMF (4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone) Aroma Component in Miso with Aging

Two kinds of miso, one with added precultured yeast and the other without, were compared with respect to the changes in the concentration of HEMF formed and the number of yeast cells in the process of aging. In miso without added yeast, the HEMF concentration increased with the increasing number of existing yeast cells. In miso without yeast aged for 21 days after the miso mash, 0.06 ppm HEMF was detected when the cell number was 2.2 × 10³ cell/g. In yeast-added miso aged for 7 days after the miso mash, no HEMF was detected, although the number of yeast cells was 1.6 × 10⁶ cell/g. In yeast-added miso aged for 14 days after the miso mash, HEMF was first detected. The pH levels of miso without yeast and with added yeast when HEMF was first detected were 5.59 and 5.57, respectively. It is suggested that the formation of HEMF in miso containing a high concentration of reducing sugar and salt was related to the growth of yeast and started when the pH level fell to less than 5.6.     

Purification and Properties of a Thermo-labile Antigen from Baker’s Yeast Cells

A thermo-labile antigen (TLA) on the yeast cell surface was isolated from a yeast cell autolyzate and purified to a homogeneous state by chromatography on an immunoadsorbent affinity column. The molecular weight of TLA was about 1.45 x 10⁵ on SDS-polyacrylamide gel electrophoresis and about 1.5 x l0⁵ on gel chromatography on Sephadex G-200. The TLA contained 74.5% protein and 25.5% sugar. It was characterized by high contents of glycine, glutamic acid, serine and aspartic acid. Half-cystine, methionine, histidine and arginine were not found. The sugar moiety was composed of galactose, mannose, N-acetylglucosamine and fucose. The antigenic determinant of TLA was distinct from that of cell wall mannan in the Ouchterlony immunodiffusion test. No precipitin line against anti-TLA serum was observed, when TLA was heated at 90°C for 10 min. Oxidation with periodate had little effect on antigenicity, but digestion with Pronase or treatment with protein denaturants resulted in loss of the antigenicity. These results suggest that the protein moiety plays an important role as the antigenic determinant of TLA. Moreover, the antiserum specific to TLA agglutinated fresh yeast cells, and the distribution of TLA was apparent on the yeast cell surface by immunofluorescence staining. These findings suggest that TLA molecules were exposed on the outer surface of the yeast cell wall.     

Effect of a phosphate solubilizing bacteria on maize growth and root exudation over four levels of labile phosphorus

The influence of inoculation with phosphate-solubilizing bacteria (Enterobacter agglomerans) on maize growth, P uptake and root exudation was studied. Plants were grown in an axenic culture device where P was supplied as soluble phosphate at different contents (0, 5, 15 or 25 ppm) in the nutrient solution and as insoluble rock phosphate added to the culture sand. Experimental device was successfully used to obtain axenic root systems or good establishment of the inoculated strain in the rhizosphere of maize (10⁹ bact. g⁻¹ dry rhizospheric material). Plant growth was promoted by inoculation only for 5 or 15 ppm of soluble P in the nutrient medium without any significant effect on P uptake by the plant, suggesting that the quantities of P released by bacterial rhizospheric activity were very small. Amounts of organic compounds (total C and water-soluble C) exuded were relatively low (3.0 to 6.4% of the total plant biomass) and were reduced by bacterial inoculation when plant growth was largely promoted. Carbon balance modification and plant growth hormone production by the inoculated strain were suspected and discussed.     

Short communication: Ethanol production from cellulose at 45°C using a batch-fed system containing alginate-immobilized Kluyveromyces marxianus IMB3

The thermotolerant yeast, Kluyveromyces marxianus IMB3, was grown in batch culture at 45°C on cellulose-containing media, supplemented with exogenous cellulase activity. At various stages during fermentation, both substrate and enzyme were added in batch mode and fermentation was continued for 220 h. Ethanol production increased to 20 g/l at 200 h, representing 45% of the maximum theoretical yield. In subsequent experiments, the organism was immobilized in calcium alginate beads and these were used in a similar, batch-fed system at 45°C. Again, fermentation was continued for 220 h and ethanol production increased to its maximum, of 28 g/l, within 100 h and this represented in excess of 60% of the maximum theoretical yield.     

High yield mixotrophic cultures of the marine microalga Tetraselmis suecica (Kylin) Butcher (Prasinophyceae)

The effects of three organic compounds were tested on one of the most used marine micro-algae in the aquaculture of molluscs and crustaceans, Tetraselmis suecica. Studies were made in axenic conditions with yeast extract, peptone and glucose added to the culture medium, each alone, in combinations of two or all together. Medium without any organic compound was used for the control. Cultures containing yeast extract grew best, reaching maximum cell density of 3.79 × 10⁶ and 3.84 × 10⁶ cells ml⁻¹. The organic carbon source affected the biochemical composition. The components most affected were the carbohydrates, with values between 6.5 pg cell⁻¹ in control cultures and 48.5 pg cell⁻¹ in glucose cultures. Protein content ranged between 27.5 pg cell⁻¹ in control cultures and 88.6 pg cell⁻¹ in yeast + glucose + peptone cultures. The lipid content changed little. Maximum protein yields were reached in cultures with yeast + glucose and with yeast - glucose - peptone, with values of 24.6 and 28.2 mg 1⁻¹ d⁻¹, respectively. These values are 22 and 25 times those in control cultures. A maximum carbohydrate yield of 7.9 mg carbohydrate per litre per day was obtained in yeast + glucose + peptone cultures, 27 times that in the control cultures. The maximum lipid yield was obtained with yeast + glucose + peptone and yeast + glucose. Maximum energy values were 308 kcal 1⁻ in yeast extract - glucose - peptone cultures and 279 kcal 1⁻¹ in yeast extract + glucose cultures. Gross energy values in control cultures were 24.5 kcal 1⁻¹, but peptone cultures presented the minimum energy value, 22 kcal 1⁻¹. The yeast extract: glucose ratio in the culture medium was optimized. A ratio 2:1 produced the best yields in cells, protein, carbohydrate and gross energy.     

Production of 21% (v/v) ethanol by fermentation of very high gravity (VHG) wheat mashes

Summary Very high gravity wheat mashes containing 300 g or more sugares per liter were prepared by enzymatic hydrolysis of starch and fermented with a commercial preparation of active dry yeast. The active dry yeast used in this study was a blend of several strains ofSaccharomyces cerevisiae. The fermentation was carried out at 20°C at different pitching rates (inoculation levels) with and without the addition of yeast extract as nutrient supplement. At a pitching rate of 76 million cells per g of mash an ethanol yield of 20.4% (v/v) was obtained. To achieve this yeast extract must be added to the wheat mash as nutrient supplement. When the pitching rate was raised to 750 million cells per g of mash, the ethanol yield increased to 21.5% (v/v) and no nutrient supplement was required. The efficiency of conversion of sugar to ethanol was 97.6% at the highest pitching rate. This declined slightly with decreasing pitching rate. A high proportion of yeast cells lost viability at high pitching rates. It is suggested that nutrients released from yeast cells that lost viability and lysed, contributed to the high yield of ethanol in the absence of any added nutrients.     

复合有机氮源对灵芝三萜液态深层发酵的影响

以灵芝为材料,在前期研究基础上,以不同酵母粉作为氮源,研究复合有机氮源对灵芝三萜液态深层发酵的影响。首先,由单因素实验考察3种不同的酵母粉对灵芝菌丝体合成灵芝三萜的影响,确定酵母粉的最适宜浓度范围。在此基础上,根据中心组合实验设计,对3种酵母粉分别采用2种复合和3种复合的方式,优化复合有机氮源的最佳组合配比。结果表明：当基础培养基中添加6.6g/L的酵母粉N-1与6.6g/L的酵母粉N-2时,灵芝三萜产量可达0.478g/L（理论产量为0.485g/L）,比添加单一酵母粉N-1、N-2、N-3分别提高了21%、139%、103%,其氮源用量为两种组合时最低。当基础培养基中酵母粉N-1、N-2与N-3添加量分别为5.07g/L、3.78g/L、7.63g/L时,灵芝三萜产量达0.514g/L（理论产量为0.510g/L）,比单因素对照组分别提高了30%、157%和74%。本研究优化的复合氮源添加方式可明显提高灵芝菌丝体液态深层发酵生产灵芝三萜的产量,为其规模化液态深层发酵的生产提供科学数据。