Anaerobic biodegradation of sugar beet pulp

Sugar beet pulp is a by-product of sugar production and consists mainly of cellulose, hemicellulose and pectin. Its composition is suitable for biological degradation. A possible alternative for the utilization of this material (besides cattle feeding) can be anaerobic methanogenic degradation. It has an additional advantage – biogas production. Beet pulp was treated by a two-step anaerobic process. The first step consisted of hydrolysis andacidification. The second step was methanogenesis. In this paper, observation ofthe process of anaerobic degradation and determination of optimal parameters is discussed. A laboratory-scale model for sugar beet pulp anaerobic biodegradation was operated. Results of model performance have shown very good pulp digestion characteristics. In addition, high efficiency removal of organic matter was achieved. Methane yield was over 0.360 m³ kg^-1 dried pulp and excess sludge production was 0.094 g per gram COD added.     

Study on the development of methanogenic microflora during anaerobic digestion of sugar beet pulp

Summary The pattern of increase in cell number in 12 different groups of bacteria was studied during anaerobic digestion of enzymatically prehydrolysed sugar beet pulp in a 70-l fermentor with sequential feeding over a period of 130 days. Glucose-fermenting bacteria accounted for 90% of the total microflora as estimated by direct epifluorescence. Strictly anaerobic bacteria were largely dominant; only 10% were methanogens. Sulphatereducing bacteria accounted for 0.1% of the total microflora. The yield of biogas was compared with the numbers of bacteria.     

Development of complex enzymatic preparations of pactinases and celulases for sugar beet marc digestion

Complex enzymatic preparations demonstrating activities homologous to pectinlyase A and heterologous to endo-1,4-β-glucanase from Penicilliumverruculosum and β-glycosidase from Aspergillusniger have been obtained on the basis of recombinant strains of the fungus Penicilliumcanescens. Two approaches were utilized: development of an enzymatic preparation on the basis of a new strain, which produced all three enzymes, and development of an enzymatic preparation via combined cultivation of three strains, each of which produced one of the enzymes.     

Single-stage, batch, leach-bed, thermophilic anaerobic digestion of spent sugar beet pulp

Spent sugar beet pulp as received was digested in a single-stage, batch, unmixed, leach-bed, laboratory scale thermophilic anaerobic digester. Biogasification of each 0.450 kg (wet weight) batch of spent pulp was initiated by inoculating with anaerobically digested liquor from previous run. The average methane yield was 0.336 m3 CH4 at STP (kgVS)(-1), the maximum methane production rate was 0.087 m3 CH4 at STP (kgVS)(-1)d(-1), average lag time to initiate methanogenesis was only 0.44 days and time required to achieve 95% methane yield was 8 days. The pH in the digesters ranged between 8.0 and 9.5. High rates of methane generation were sustained even at high pH values. The equivalent organic loading rate in the batch digesters was 4 kgCODm(-3)d(-1). The digestion process used here offers significant improvements over one-stage and two-stage systems reported in the literature with comparable performance as it is a single-stage system where the feedstock does not require size reduction, and mixing is not required in the digester.     

Phenotypic polymorphism and allele differentiation of isozymes in fodder beet,multigerm sugar beet and monogerm sugar beet

Summary Thirteen enzymes (MDH, SDH, LAP, PGM, PX, IDH, GPI, 6PGD, APH, GOT, GDH, ME and SOD) of 3 cultivated beet (B. vulgaris L.) gene pools, comprising 12 accessions of fodder beet, 11 of old multigerm sugar beet and 10 of modern monogerm sugar beet, were investigated using horizontal starch gel electrophoresis. Eleven accessions of primitive or wild B. vulgaris were also included for the comparison of isozymes. Variation in isozyme phenotypes was investigated to detect diversity in the three cultivated forms of beet. Phenotypic variation was observed in all except ME and SOD, which were monomorphic. A high degree of phenotypic polymorphism (Pj) was found in GDH, PGM, IDH, APH and MDH. Differences in phenotypic polymorphism in MDH, GPI and PX were recognized between fodder beet and both sugar beet groups. Average polymorphism for 13 enzymes in both sugar beets was significantly higher than that in fodder beet. For 13 enzymes, the existence of high isozyme diversity in both sugar beet gene pools was revealed. Allele frequencies in 13 alleles of five enzyme-coding loci, Lap, Px-1, Aph-1, Got-2 and Gdh-2, were investigated. New alleles, Px-1 ¹ and Got-2 ¹, were found in fodder beet accessions. No significant differences of average allele frequencies of five loci between fodder beet and both sugar beets were recognized. Several unique alleles and different isozyme phenotypes were observed in the accessions of B. vulgaris ssp. macrocarpa and ssp. adanensis. Future utilization of cultivated beet gene pools for sugar beet breeding is discussed from the viewpoint of genetic resources.     

Bird damage to sugar beet

Pest damage to sugar beet, including that by birds, has been recorded since 1957. During that time damage by rooks has decreased almost to nil, but some other bird damage has greatly increased, most probably as a result of changing agronomic practices, especially the extensive use of herbicides, the introduction of monogerm seed, and the increasing practice of ‘planting-to-a-stand’. The most severe bird damage in the spring is grazing by several species, and in early summer localized felling of plants by pheasants. Observations in the mid–1960's of causes of seedling and plant losses suggested that birds were then of minor importance; the British Sugar Corporation currently consider that birds are the most serious pest of sugar beet. The distribution of the reported damage does not seem to follow any national pattern. In small-plot field trials possible repellent materials such as anthraquinone, methiocarb or thiram, applied to seed or foliage, did not decrease the extent of grazing.     

Reducing the risk of foaming and decreasing viscosity by two-stage anaerobic digestion of sugar beet pressed pulp

Anaerobic digestion (AD) of sugar beet pressed pulp (SBPP) is a promising treatment concept. It produces biogas as a renewable energy source making sugar production more energy efficient and it turns SBPP from a residue into a valuable resource. In this study one- and two-stage mono fermentation at mesophilic conditions in a continuous stirred tank reactor were compared. Also the optimal incubation temperature for the pre-acidification stage was studied. The fastest pre-acidification, with a hydraulic retention time (HRT) of 4 days, occurred at a temperature of 55 °C. In the methanogenic reactor of the two-stage system stable fermentation at loading rate of 7 kg VS/m³ d was demonstrated. No artificial pH adjustment was necessary to maintain optimum levels in both the pre-acidification and the methanogenic reactor. The total HRT of the two-stage AD was 36 days which is considerably lower compared to the one-stage AD (50 days). The frequently observed problem of foaming at high loading rates was less severe in the two-stage reactor. Moreover the viscosity of digestate in the methanogenic stage of the two-stage fermentation was in average tenfold lower than in the one-stage fermentation. This decreases the energy input for the reactor stirring about 80 %. The observed advantages make the two-stage process economically attractive, despite higher investments for a two reactor system.     

Enzymic saccharification of sugar beet pulp

Culture filtrates of Talaromyces emersonii UCG 208 grown on beet pulp can convert the polysaccharide components of this agricultural waste to soluble sugars. The saccharification process is facilitated if the pulp is milled or incubated with alkali or peracetic acid before addition of enzyme. However, treatment of unmilled pulp with commercial pectinase prior to incubation with Talaromyces filtrate is also very effective; under suitable conditions, complete hydrolysis of total polysaccharides has been achieved.     

Production of regenerants in sugar beet

The experimental results are presented on production of plants-regenerants of the sugar beet (Beta vulgaris L.) via callus formation or direct organogenesis from the leaf tissues. The method of aseptic treatment was developed for the seeds with strong bacterial and fungal invasion. The regenerant plants were obtained in the presence of various concentrations of synthetic hormones, such as cytokinin (6-benzaminopurine) and auxin (naphthylacetic acid), and inhibitor of auxin transport in plants (2,3,5-triiodobenzoic acid). This combination of growth regulators made it possible to avoid callus formation. The genotype of initial plants affected the capacity for callus formation and regeneration. The temperature influenced rhizogenesis in regenerants.     

Enzymatic hydrolysis of sugar beet pulp

Summary In order to prepare fermentable sugars from cellulosic raw materials, we have tested the effect of different pretreatments (heat, chemicals, pectinase) on enzymatic hydrolysis of sugar beet pulp. Best results (90% hydrolysis) have been obtained after heat treatment (30 mn at 120°C) with 16 IU.Fp.g^–1 of fresh pulp in 48 hours.     

Volatile components of sugar beet leaves

Extracts of both young and old sugar beet plants were obtained using a modified Likens and Nickerson apparatus. Constituents were identified by GC/MS, and using selected ion monitoring it was shown that the previously determined phenylacetonitrile was probably not of glucosinolate origin. Some unsaturated aldehydes, alcohols and derivatives (enzymic lipid degradation products) were formed to greater extents by the younger leaves, but otherwise such quantitative differences were relatively few and generally random. An interesting range of chlorinated compounds was obtained only from the older plants; a pesticide origin is suggested.     

Adenylate patterns of autumn-sown sugar beet differ from spring-sown sugar beet. Implications for root quality

Sugar beet ( Beta vulgaris L) is generally cultivated using two different planting and harvest patterns. In northern zones, spring sugar beet is sown in spring and harvested in autumn, whereas in subtropical latitudes, autumn sugar beet is sown in autumn and harvested in summer. The industrial quality of the root is frequently higher in spring-sown sugar beet crops. In order to explore physiological changes associated with this fact, this study has been focused on the seasonal changes of adenosine 5'-triphosphate and adenosine 5'-diphosphate levels in the storage roots of sugar beet plants, as an index of its metabolic status. The results obtained correspond to a different metabolic status of spring and autumn sugar beet at the moment of harvest. The adenylate patterns of autumn beets suggested a functional and active respiratory system. On the contrary, the patterns shown by spring beets corresponded to those we would expect to see in plants becoming dormant. The proline and glucose contents, which decrease the industrial quality of the root, and the respiratory rate measured in autumn-sown sugar beets, were nearly twice those of spring-sown sugar beets. The combination of an active respiratory system, which allows the carbohydrate catabolism and the synthesis of stress molecules, with the environmental factors at the time of the harvest, could be the underlying physiological mechanism causing some of the differences between spring- and autumn-sown sugar beet crops.     

Occurrence of beet western yellows virus in sugar beet in Czechoslovakia

The beet western yellows virus (BWYV) was identified in sugar beet plants with leaf yellowing symptoms. When transmitted toSinapis alba L. the virus isolate caused severe symptoms of yellowing and violetting of the interveinal leaf tissue of this plant. By aphidsMyzus persicae (Sulz.) the virus isolate was transmitted toLactuca sativa L.,Raphanus sativus L. var.radicula Pers.,Baphanus sativus L. ssp.sativus L. ap., and toBrassica oleracea L. var.gemmifera DC. InLactuca sativa plants the virus induces a yellowing along with thickenning and brittleness of leaves and with mild dwarfing of the plants. InBaphanus sativus var.radicula andBaphanus sativus ssp.sativus plants it brings about a yellowing of the leaf margins with a change in consistency as was the case in lettuce, and inBrassica oleracea var.gemmifera it causes violet spots on the lower leaf sides. The transmission was proved in repeated experiments by a backtransmission to beet andSinapis alba and further transmission from beet toSinapis alba. The transmission of the virus isolate toVicia faba L.,Datura stramonium L., andPetunia hybrida hort. was unsuccessful. In the course of transmissions the isolate properties did not change. In its host range the virus resembles the Duffus’ strain 3 BWYV, isolated from beet in the U.S.A. This is the first characteristic of an Europian BWYV isolate, as obtained from naturally infected beet plants.     

Mapping sugar beet pectin acetylation pattern

Homogalacturonan-derived partly methylated and/or acetylated oligogalacturonates were recovered after enzymatic hydrolysis (endo-polygalacturonase+pectin methyl esterase+side-chain degrading enzymes) of sugar beet pectin followed by anion-exchange and size exclusion chromatography. Around 90% of the GalA and 75% of the acetyl groups present in the initial sugar beet pectin were recovered as homogalacturonan-derived oligogalacturonates, the remaining GalA and acetyl belonging to rhamnogalacturonic regions. Around 50% of the acetyl groups present in sugar beet homogalacturonans were recovered as partly methylated and/or acetylated oligogalacturonates of degree of polymerisation 5 whose structures were determined by electrospray ionization ion trap mass spectrometry (ESI-IT-MSn). 2-O-acetyl- and 3-O-acetyl-GalA were detected in roughly similar amounts but 2,3-di-O-acetylation was absent. Methyl-esterified GalA residues occurred mainly upstream 2-O-acetyl GalA. Oligogalacturonates containing GalA residues that are at once methyl- and acetyl-esterified were recovered in very limited amounts. A tentative mapping of the distribution of acetyl and methyl esters within sugar beet homogalacturonans is proposed. Unsubstituted GalA residues are likely to be present in limited amounts (approximately 10% of total GalA residues), due to the fact that methyl and acetyl groups are assumed to be most often not carried by the same residues.     

Electron microscopical proof of the presence of sugar beet yellows virus particles in the roots of sugar beet

The presence of beet yellows virus (BYV) particles was electron microscopically proved in the roots of sugar beet. Specimens for the electron microscopical examination of root sap were prepared by differential centrifugation. It was proved that, contrary to expectations, examinations in spring showed most virus particles in the basal part of the root. At the same time it was found by experiment that the diagnostical BYV antiserum, for which the antigen was prepared from sugar beet leaves, did not react with a purificate of BYV containing virus particles.