Extracellular prolyl endoprotease from Aspergillus niger and its use in the debittering of protein hydrolysates

The observation that the bitterest peptides from casein hydrolysates contain several proline residues led us to hypothesize that a proline-specific protease would be instrumental in debittering such peptides. To identify the desired proline-specific activity, a microbiological screening was carried out in which the chromogenic peptide benzyloxycarbonyl-glycine-proline-p-nitroanilide (Z-Gly-Pro-pNA) was used as the substrate. An Aspergillus niger (A. niger) strain was identified that produces an extracellular proline-specific protease with an acidic pH optimum. On the basis of sequence similarities, we conclude that the A. niger-derived enzyme probably belongs to the S28 family of clan SC of serine proteases rather than the S9 family to which prolyl oligopeptidases belong. Incubating the overexpressed and purified enzyme with bitter casein hydrolysates showed a major debittering effect. Reversed phase HPLC analysis revealed that this debittering effect is accompanied by a significant reduction of the number of hydrophobic peptides present.     

Characterizing the soil moisture regime of a Typic Haplohumod

The saturated hydraulic conductivities (K_sat) of 22 spodic horizons with and without a thin iron pan, were measured in situ with a new technique, using large, carved-out columns, encased in gypsum. Measured infiltration rates and pressure heads above and below the spodic horizons allowed calculation of K_sat, which averaged 8 cm d^?1. Flow rates averaged 32 cm d^?1, however, due to a hydraulic head gradient across the spodic horizon of 4 cm cm^?1. Occurrence of a thin iron pan in the spodic horizon did not affect its K_sat-value. The measured high flow rates exclude the occurrence of perched watertables. Lateral flow of water, forming surface ponds in local depressions, was due to surface runoff, rather than to lateral movement of perched water: surface ponding of water occurred also in soils in which the spodic horizon had been mixed by tillage.     

Obtaining soil physical field data for simulating soil moisture regimes and associated potato growth

Soil moisture regimes under potatoes were monitored during two growing seasons in a sandy loam and a silty clay loam soil in The Netherlands. Measured moisture contents were used to validate those calculated using the simulation model SWATRE. The model requires hydraulic conductivity, moisture retention, rooting-depth and soil cover data, which were obtained in this study. Measured groundwater levels formed the lower boundary condition and precipitation and potential evapotranspiration the upper boundary condition of the model flow system. Calculated moisture contents agreed well with measured values, but only when in situ moisture retention curves were used and when the effects of cracking in the silty clay loam soil were expressed by modifying the ?-θ function. The moisture supply capacity of the sandy loam soil was highest, particularly in the first growing season and this was interpreted as a major reason for the corresponding higher yields. Simulation can be used for predicting the soil moisture supply capacity in the context of land evaluation, when soil-cover and rooting depth are standardized, for example by simulating growth of a grass crop.     

Movement and conversion of aldicarb and its oxidation products in potato fields

In Spring, the insecticide and nematicide aldicarb in the granular formulation Temik 10G (supplied by Union Carbide) was broadcast on three potato fields and incorporated by rotary cultivation. Ridges were formed by repeated runs with ridging implements. The soil was sampled in layers of 0.1 m up to 0.8 m depth three to four times during the growing season. Aldicarb itself was almost completely converted on a humic sand soil and two loam soils within one month. During the growing season, its sulphoxide and sulphone, which are toxic and are formed by oxidation, were retained mainly in the top 0.3 m of all three soils. Relatively high concentrations were measured only in the top 0.2 m, indicating limited redistribution by leaching.Low to very low contents were found up to 0.8 m depth especially on one of the loam soils where the highest total rainfall was measured from May to October (328 mm). In a humic sand soil, leaching in the furrow was deepter than below the ridge. At the end of the growing season, the sulphoxide plus sulphone corresponded to a mass fraction of 5.7 to 6.7% of the dosage in the two loam soils and to 17% in the humic sand soil. These residues were mainly concentrated in the centre of the ridges.Samenvatting In het voorjaar werd op drie aardappelvelden het insecticide en nematicide aldicarb als Temik 10G breedwerpig toegediend en met een frees ingewerkt. Ruggen werden opgebouwd door herhaalde bewerkingen met aanaardwerktuigen. Gedurende het groeiseizoen werd de grond drie tot viermaal bemonsterd in laagjes van 0,1 m tot op 0,8 m diepte. Aldicarb zelf was binnen een maand bijna geheel omgezet zowel in een humeuze zandgrond als in twee zavelgronden. Gedurende het groeiseizoen bleven het sulfoxide en het sulfon, welke produkten eveneens toxisch zijn en door oxidatie van aldicarb worden gevormd, voornamelijk in de toplaag van 0,3 m bij alle drie gronden. Relatief hoge concentraties werden alleen gemeten in de bovenste 0,2 m, wat wijst op een beperkte herverdeling door uitspoeling.Lage tot zeer lage gehalten werden tot 0,8 m diepte gevonden, met name in één van de zavelgronden waar ook de grootste hoeveelheid neerslag van mei tot oktober (328 mm) werd gemeten. In een humeuze zandgrond was de uitspoeling in de voor groter dan die beneden de rug. Aan het eind van het groeiseizoen kwam de hoeveelheid resterend sulfoxide plus sulfon in de twee zavelgronden overeen met een massafractie van 5,7 tot 6,7% van de dosering en in de humeuze zandgrond met 17%. Deze residuen waren voor het grootste deel geconcentreerd in het centrum van de ruggen.     

Sensitivity of Ustilago maydis to pyrazophos and one of its conversion products,and failure to induce resistance with UV-treatment

2-Hydroxy-5-methyl-6-ethoxycarbonylpyrazolo(1,5-a)pyrimidine (PP), a conversion product of pyrazophos, shows considerable toxicity toUstilago maydis, when administered to this fungus in a solution at pH<5. Evidence was obtained thatU. maydis may convert pyrazophos in to PP, and that the latter compound is the toxic principle responsible for the action of pyrazophos. By UV-irradiation of sporidia no PP-resistant mutants were obtained. This does not support the hypothesis that this toxicant acts by specific inhibition of one enzyme system.     

Decreased sensitivity to pyrazophos of cucumber and gherkin powdery mildew

In some cucumber and gherkin greenhouses in the Netherlands, where Curamil (a.i. pyrazophos) had been used for control of powdery mildew, unsatisfactory disease control was obtained in recent years. Laboratory tests revealed a decreased sensitivity of the pathogen to the fungicide. The fitness and the competitive ability of pyrazophos resistant strains in absence of the fungicide appeared to be somewhat lower than that of the normal, sensitive strains. The implications with respect to disease control in practice are discussed.Samenvatting In enkele kassen, waar Curamil (a.i. pyrazofos) in 1977 aangewend werd ter bestrijding van meeldauw op komkommers en augurken, werd onvoldoende effekt met dit middel tegen deze ziekte verkregen. Toetsing in het laboratorium bracht een verminderde gevoeligheid van het pathogeen voor het fungicide aan het licht (Tabel 1). Bij afwezigheid van pyrazofos bleek het competitief vermogen (Tabel 2) en de vitaliteit (Tabel 3) van de pyrazofos resistente isolaten wat lager te zijn dan die van het normale, gevoelige pathogeen. De betekenis hiervan met betrekking tot de ziektebestrijding wordt besproken.     

The composition of adipose tissue and its usefulness as a parameter for carcass composition in growing pigs

Growing pigs, varying in body weight between 25 and 110 kg, showed differences in fat content between adipose tissues from different body sites. In all adipose tissues fat content increased during growth; the increase in fat content was, as shown for backfat, affected by the level of daily energy intake. It is concluded that the dry matter (DM) or fat content in a single sample of adipose tissue cannot be used as an accurate parameter for fatness of carcasses at a normal slaughter weight.     

Oort en het onderwijs

Summary When Oort took the chair of plant pathology in 1949 his teaching included virology and nematology in addition of fungal and bacterial diseases. In 1950 a chair of plant virology was founded while in 1956 a lecturer in plant nematology was added to the chair of phytopathology. Only then could Oort concentrate on fungal diseases. Oort closely followed the turbulent evolution of phytopathology and, though he surveyed a wide field, he did not hesitate to invite staff members to teach special courses. In recent years junior students had to follow courses in Introductory Plant Pathology, General Plant Pathology, Morphology and Taxonomy of Fungi, and in addition a practical course in symptom analysis and morbid anatomy. For advanced students seminars were organized, each concentrating on one topic, e.g. Resistance or Soil Borne Diseases. Moreover, the advanced students had to follow a practical course in phytopathological and pathophysiological techniques. Specialized practical courses on Epidemiology and Resistance and Physiology of parasitism were started recently. The students usually did for 3 or 6 months research work under the guidance of a staff member. A research report and a literature review on a chosen topic were obligatory for all advanced students. Oort's keen interest in teaching aids led to the accumulation of a large slide collection which contained hundreds of colour slides of disease symptoms, and to a collection of diseased plants in permanent mounts. In nearly 20 years Oort accepted and discussed over 300 research reports, about 225 literature reviews, 21 doctor's theses and examined over 700 students.