Interrelation between penicillin productivity and growth rate

Summary Fermentations were carried out in an 801 tower-loop reactor with pellets of Penicillium chrysogenum. The development of the inner structure of the pellets with regard to various fermentation conditions was observed by means of histological preparations of the pellets. Under conditions of energy-source-limitation mycelial tip growth and lysis of other mycelial parts exist simultaneously. Thus the net growth rate (formation rate of cell mass) is higher than the gross growth rate (multiplication rate of cell mass). Under conditions of nitrogen limitation, gross growth rate and net growth rate are identical. A very strict correlation between gross growth rate and penicillin production rate was found as long as sufficient oxygen supply could be maintained and carbon catabolite repression was avoided. The energy source requirement of the biomass can be described with the sum of three terms that correspond to gross growth, lysis compensation growth and maintenance.Symbols a Constant 1/l h - b Constant - K Decay rate constant for product 1/h - K ₁ Substrate inhibition constant g/l - K _op Controls saturation constant for oxygen g/l - K _p Saturation constant for substrate g/l - m Maintenance coefficient 1/h - ms Apparent maintenance coefficient 1/h - O Dissolved oxygen concentration g/l - P Product concentration g/l - p Exponent of O - q Specific productivity 1/h - S Substrate concentration g/l - t Time h - t ₁ Beginning of production phase h - t ₂ Time of pellet dissolution h - V Liquid volume of fermentation broth l - X Dry cell mass concentration g/l - Y Yield of dry cell mass from energy substrate - g Specific gross growth rate of biomass 1/h - l Specific lysis rate of cell mass 1/h - n Specific net growth rate of cell mass 1/h - p Maximum specific rate of product formation 1/h     

Cyclic-AMP-dependent protein kinase type II is associated with the Golgi complex and with centrosomes

The subcellular distribution of the type II enzyme of cAMP-dependent protein kinase (cAMP-dPK II) in epithelial and fibroblastic cells was determined by indirect immunofluorescence microscopy. In interphase cells both regulatory (R II) and catalytic (C) subunits were concentrated in a perinuclear area. By comparison of the R II distribution with the location of a bona fide Golgi membrane constituent, this area was identified as the Golgi complex. The cytochemical localization of R II was confirmed by subcellular fractionation. In addition, cAMP-dPK II was associated with microtubule-organizing centers, in particular with mitotic spindle poles. These distributions of cAMP-dPK II probably represent important factors in mediating the effects of cAMP on basic cellular activities ranging from secretion and proliferation to cell shape and motility.     

Fortpflanzung und entwicklung vonAnemonia sulcata (Anthozoa,Actiniaria) II. Frühentwicklung,Blastula und Gastrula

Early developmental stages of theAnemonia germ are characterized by asynchronously dividing nuclei and an extreme delay of blastomere differentiation. The nuclei migrate to the periphery, whereas nutritive substances remain in the interior. Following this stage, the appearance of cell boundaries results in the formation of the blastoderm and the simultaneous division of the yolk into many fragments. Most of them are exclusively filled with reserve material; only very few contain zooxanthellae or nuclei. On the embryo's surface, conically shaped bundles of long microvilli are obvious. They appear to be less regularly arranged than the spines of oocytes before insemination. Pigment granules that have originated from fusing Golgi vesicles are crowded peripherally in the blastoderm cells. In the nucleoplasm single annulate lamellae that seem to be cut off from the nuclear envelope can frequently be found. There is no further cellular differentiation until gastrulation is completed. Though yolk-containing cellular fragments occupy nearly the whole blastocoel, entoderm formation occurs by invagination. Ultrastructural observations provide evidence of the existence of interstices between entoderm cells that allow all nutritive substances to pass gradually into the gastric cavity. In the region of the blastoporus there are cellular processes enveloping reserve material. Presumably, these observations indicate a so-called filtration of nutritive yolk (Korschelt & Heider, 1909) that might represent an additional mode for the transfer of yolk-containing cellular fragments from blastocoel into gastrocoel.     

Floristic groups and plant communities of southeastern Santa Fe,Argentina

This paper is a survey of the vegetation of the southeastern departments in the Province of Santa Fe (Argentina). The vegetation was analyzed following Braun-Blanquet's approach modified by Mueller-Dombois & Ellenberg (1974). The most relevant species of the region were placed in 25 groups according to their requirements or general behaviour. Most of the communities are herbaceous, and apart from the woody and some other miscellaneous ones they were grouped into three ecologically and floristically defined sets. The first set, the Stipa grasslands and related communities, which are characterized by the more or less abundant presence of Stipa hyalina, Stipa neesiana and Stipa papposa, comprises five different communities. The second, the halophilous communities, comprises five communities, the two Spartina ssp. grasslands, the halophilous prairies of Distichlis spicata, the short sedge Scirpus americanus communities and the ‘pela-dales’. The third, the hygrophilous communities, comprises nine communities which are not so well defined as the ones in the other sets. Besides, two further communities have been included, the Paspalum quadrifarium and the Melica macra tall grasslands.     

Isolation of acid-resistant urinary trypsin inhibitors by high performance liquid chromatography and their characterization by N-terminal amino-acid sequence determination

Two crude fractions of acid-resistant trypsin inhibitors (apparent molecular masses 44 and 20 kDa, respectively) were prepared from human urine by gel permeation chromatography. From both preparations the pure inhibitors were isolated by high performance liquid chromatography (HPLC). Their N-terminal amino-acid sequences were determined and compared with those of HI-30 and HI-14 as isolated by reversible binding to either immobilized trypsin or immobilized chymotrypsin. The N-terminal amino-acid sequence of the high-molecular mass inhibitor UI-I isolated by HPLC was identical with those of HI-30 and UI-C-I isolated via immobilized trypsin or chymotrypsin, respectively. The low-molecular mass inhibitors UI-II and UI-C-II differ from HI-14 by the N-terminal extension Glu-Val-Thr-Lys-when obtained by HPLC or by the extension Thr-Lys-when obtained via immobilized chymotrypsin, respectively. The comparison of these N-termini with the amino-acid sequence of HI-30 (Ala1-...-Val16-Thr-Glu-Val-Thr-Lys-HI-14) defines the low molecular urinary trypsin inhibitors as proteolytic degradation products of the high-molecular urinary inhibitor. Proteolysis may occur at different bonds. The existing discrepancies in molecular architecture and in molecular masses of the urinary trypsin inhibitors are discussed.     

Purification and properties of a stilbene synthase from induced cell suspension cultures of peanut

Stilbene synthase ( resveratrol -forming) converts one molecule of rho- coumaroyl -CoA and three molecules of malonyl-CoA into 3,4',5- trihydroxystilbene . Following selective induction of stilbene synthesis in cell suspension cultures of peanut (Arachis hypogaea), the enzyme was extracted and purified to apparent homogeneity by chromatography on DEAE-cellulose and hydroxylapatite. The enzyme was found to be a dimer of estimated Mr = 90,000 exhibiting under denaturing conditions a subunit Mr of approximately 45,000. The isoelectric point was determined with pI = 4.8. The enzyme's high selectivity towards rho- coumaroyl -CoA (Km = 2 microM) as substrate qualified it as resveratrol -forming stilbene synthase. Structurally related CoA esters, e.g. dihydro-rho- coumaroyl -CoA and cinnamoyl-CoA, were converted less than 1/10 as efficiently as rho- coumaroyl -CoA. Malonyl-CoA (Km = 10 microM) could not be substituted by acetyl-CoA. The purified enzyme was free of chalcone synthase activity. Antibodies raised against stilbene synthase were shown to be monospecific and not to cross-react with chalcone synthase.     

Penicillin acylase production by E. coli

Enzyme production with E. coli ATCC 11105, in a complex medium using phenylacetic acid as inducer is carried out in a stirred-tank reactor of 10 dm³ and an airlift tower-loop reactor of 60 dm³ with outer loop at a temperature of 27 °C. The optimum inducer concentration was 0.8 kg/m³, which was kept constant by fed-batch operation. The optimum of the relative dissolved O₂-concentration with regard to saturation is below 10% in a stirred-tank reactor and at 35% in a tower-loop reactor. It was kept constant by parameter-adaptive control of the aeration rate. In a stirred-tank enzyme productivity is slightly higher than in a tower-loop reactor, and much higher than in a bubble column reactor.List of Symbols CPR kg/(m³ h) CO₂-production rate - OTR kg/(m³ h) O₂-transfer rate - OUR kg/(m³ h) O₂-utilization rate - PAA phenylacetic acid (inducer) - RQ = CPR/OUR respiratory quotient - X kg/m³ cell mass concentration - _m h^–1 maximum specific growth rate     

Growth of E. coli in a stirred tank and in an air lift tower reactor with an outer loop

E. coli ATCC 11105 was cultivated in a 10-1 stirred tank reactor and in a 60-1 tower loop reactor in batch and continuous operation. By on-line measurements of O₂ and CO₂ concentrations in the outlet gas, pH, temperature, cell mass concentration X as well as dissolved O₂ concentration along the tower in the broth, gas holdup, broth recirculation rate through the loop and by offline measurements of substrate concentration DOC and cell mass concentration along the tower, the maximum specific growth rate _m , yield coefficients Y _X/S. Y _X/DOC and were evaluated in stirred tank and tower loop in batch and continuous cultures with and without motionless mixers in the tower and at different broth circulation rates through the loop. To control the accuracy of the measurements the C balance was calculated and 95% of the C content was covered.The biological parameters determined depend on the mode of operation as well as on the reactor used. Furthermore, they depend on the recirculation rate of the broth and built-ins in the tower. The unstructured cell and reactor models are unable to explain these differences. Obviously, structured cell and reactor models are needed. The cell mass concentration can be determined on line by NADH fluorescence in balanced growth, if the model parameters are determined under the same operational conditions in the same reactor.List of Symbols a, b empirical parameters in Eq. (1) - CPR kg/(m³ h) CO₂ production rate - C kg/m³ concentration - D l/h dilution rate - DOC kg/m³ dissolved organic carbon - I net. fluorescence intensity - K _S kg/m³ Monod constant - k _L a l/h volumetric mass transfer coefficient - OTR kg/(m³ h) oxygen transfer rate - OUR kg/(m³ h) oxygen utilization rate - RQ = CPR/OUR respiratory quotient - S kg/m³ substrate concentration - t h,min, s time - t _u min recirculation time - t _M min mixing time - v m³/h volumetric flow rate through the loop - X kg/m³ (dry) cell mass concentration - Y _X/S yield coefficient of cell mass with regard to the consumed substrate - Y _X/DOC yield coefficient of the cell mass with regard to the consumed DOC - Y _X/O yield coefficient of the cell mass with regard to the consumed oxygen - Z relative distance in the tower from the aerator with regard to the height of the aerated broth - l/h specific growth rate - _m l/h maximum specific growth rate Indices f feed - e outlet