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EFFECTS OF ASPARTATE ON GROWTH AND ON THE SYNTHESIS OF ALPHA-AMYLASE IN PSEUDOMONAS SACCHAROPHILA.

Eisenstadt, Jerome M. (Brandeis University, Waltham, Mass.), and Harold P. Klein. Effects of aspartate on growth and on the synthesis of alpha-amylase in Pseudomonas saccharophila. J. Bacteriol. 87:1355-1363. 1964.-Protein synthesis by starch-grown cells of Pseudomonas saccharophila is inhibited by low concentrations of d-aspartate, whereas assimilation continues for several hours under these conditions. These effects are also observed when the cells are grown with cellobiose or maltose, but not with a wide variety of other carbon sources. l-Aspartate at a ratio of about 6:1 completely reverses these effects, whereas adenosine partially does so. One possible basis of the inhibitory effects of d-aspartate is in the synthesis of adenosine from inosine.

Amylases↗

UTILIZATION OF AROMATIC AMINO ACIDS BY HYDROGENOMONAS FACILIS.

DeCicco, B. T. (Rutgers, The State University, New Brunswick, N.J.), and W. W. Umbreit. Utilization of aromatic amino acids by Hydrogenomonas facilis. J. Bacteriol. 88:1590-1594. 1964.-An auxotrophic mutant of Hydrogenomonas facilis was isolated which requires tryptophan, phenylalanine, and p-aminobenzoic acid (PABA) for growth. With glucose as the main carbon and energy source, the quantitative requirements for tryptophan and PABA were at normal microgram levels, but the requirement for phenylalanine was very large and approached substrate concentrations. The large phenylalanine requirement is due to a rapid oxidation and degradation of phenylalanine by the mutant. The utilization of both phenylalanine and glucose is adaptive, and the presence of phenylalanine partially inhibits the induction of the glucose-utilizing system. Wild-type H. facilis can utilize either phenylalanine or tyrosine for growth. Tracer studies indicated that during growth on phenylalanine, the aromatic ring is opened and degraded. Wild-type cells grown on either phenylalanine or tyrosine can oxidize phenylalanine, tyrosine, or phenylpyruvate without a lag. Another inducible pathway enables H. facilis to utilize either quinate or 3,4-dihydroxybenzoate for growth, and sequential adaptation studies revealed that quinate is converted to 3,4-dihydroxybenzoate during its degradation. Mutants may be obtained which can also utilize 2,5-dihydroxybenzoate for growth.

4-Aminobenzoic Acid↗

CHARACTERIZATION OF POLY-BETA-HYDROXYBUTYRATE EXTRACTED FROM DIFFERENT BACTERIA.

Lundgren, D. G. (Syracuse University, Syracuse, N.Y.), R. Alper, C. Schnaitman and R. H. Marchessault. Characterization of poly-beta-hydroxybutyrate extracted from different bacteria. J. Bacteriol. 89:245-251. 1965.-Poly-beta-hydroxybutyrate (PHB) from different bacterial genera was studied with regard to its crystal structure, infrared absorption, intrinsic viscosity, and electron microscopy. All PHB samples precipitated from dilute chloroform solution gave identical X-ray diffractograms confirming uniformity of crystal structure, and uniformity of molecular structure, based on the similarity of the recorded infrared spectra, was also established. Crystal morphology was also similar, showing the reported "lath" shape structure for purified polymer from Bacillus cereus. Intrinsic viscosity ranged from 0.04 to 11.5 depending upon the polymer treatment; polymer molecular weights, based upon viscometry, could be estimated to range from 1,000 to 250,000. It is concluded that the same basic molecule is involved in all PHB present in the bacterial kindgom.

Azotobacter↗

CHARACTERISTICS AND INTERMEDIATES OF SHORT-TERM C-14-O-2 INCORPORATION DURING RIBOSE OXIDATION BY HYDROGENOMONAS FACILIS.

McFadden, B. A. (Washington State University, Pullman), and H. R. Homann. Characteristics and intermediates of short-term C(14)O(2) incorporation during ribose oxidation by Hydrogenomonas facilis. J. Bacteriol. 89:839-847. 1965.-Ribose-grown cells of Hydrogenomonas facilis, which had been suspended in growth medium and were oxidizing ribose, were exposed to HC(14)O(3) (-) of high specific activity. The uptake was proportional to cell mass. Short-term uptake (less than 2 min) was completely inhibited by 10(-3)m 2,4-dinitrophenol (DNP) or by <4 x 10(-6)mm-chlorocarbonyl cyanide phenylhydrazone, and to the extent of 42% by 5 x 10(-5)m DNP. The following observations were made in kinetic studies (8, 16, 35, 67, 96, and 181 sec) of fixation in the presence of ribose. Glutamate was extensively labeled in periods up to 3 min. It was one of the major early products, containing 30% of the label at 8 sec. The sugar phosphate fraction was not detectably labeled at 8 or 16 sec, but its C(14)-content increased rapidly to 27% at 35 sec and then slowly decreased. Label in phosphoglycerate, phosphoenolpyruvate, and alanine did not appear until 35 sec, and did not exceed about 7, 2, and 3%, respectively, of the total extracted radioactivity. Adenosine triphosphate and adenosine diphosphate were heavily labeled after fixation in a pilot study for 125 sec. Although considerable radioactivity incorporated during the pilot study was intractable by the extraction procedure employed, virtually no C(14) was found in the residue in poly-beta-hydroxybutyric acid. A large number of amino acids and organic acids and some organic phosphates were not detectably labeled in any of the experiments. Omission of ribose greatly diminished incorporation, particularly into glutamate.

2,4-Dinitrophenol↗

NICKEL-DEPENDENT CHEMOLITHOTROPHIC GROWTH OF TWO HYDROGENOMONAS STRAINS.

Bartha, R. (University of Washington, Seattle), and E. J. Ordal. Nickel-dependent chemolithotrophic growth of two Hydrogenomonas strains. J. Bacteriol. 89:1015-1019. 1965.-The trace element requirements for growth of facultative chemolithotrophic Hydrogenomonas strains H1 and H16 were investigated under both autotrophic and heterotrophic conditions. The organisms were grown in a mineral medium, rendered deficient in trace elements by extraction with 8-hydroxyquinoline and chloroform, and, in some cases, by coprecipitation with copper. The organic substrates, succinate and fumarate, used for heterotrophic growth were treated in a similar fashion. Acetate and butyrate were purified by redistillation. It was found that iron alone was required for heterotrophic growth (optimal concentration, 1.5 x 10(-6)m Fe(+++)), but cells grown chemolithotrophically on molecular hydrogen required the addition of nickel. The yield of protein was proportional to the nickel added, reaching a maximum at 3 x 10(-7)m Ni(++). Manganese, cobalt, copper, and zinc, alone or in combination, failed to substitute for nickel in the experiments with Hydrogenomonas. Although nickel is required specifically for the chemolithotrophic growth of Hydrogenomonas, nickel deficiency did not affect: (i) the synthesis or activation of hydrogenase, (ii) the Knallgas reaction, (iii) the assimilation of CO(2) by resting cells, or the synthesis of the storage material poly-beta-hydroxybutyric acid. It is suggested that nickel participates in some reaction involved in CO(2) fixation by growing cells.

Acetates↗

Evaluation of nitrate removal by continuous culturing of an aerobic denitrifying bacterium, Paracoccus pantotrophus.

Nitrate removal under aerobic conditions was investigated using pure cultures of Paracoccus pantotrophus, which is a well-known aerobic-denitrifying (AD) bacterium. When a high concentration of cultures with a high carbon/nitrogen (C/N) ratio was preserved at the beginning of batch experiments, subsequently added nitrate was completely removed. When continuous culturing was perpetuated, a high nitrate removal rate (66.5%) was observed on day 4 post-culture, although gradual decreases in AD ability with time were observed. The attenuation in AD ability was probably caused by carbon limitation, because when carbon concentration of inflow water was doubled, nitrate removal efficiency improved from 18.1% to 59.6%. Bacterial community analysis using the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method showed that P. pantotrophus disappeared in the suspended medium on day 8 post-culture, whereas other bacterial communities dominated by Acidovorax sp. appeared. Interestingly, this replaced bacterial community also showed AD ability. As P. pantotrophus was detected as attached colonies around the membrane and bottom of the reactor, this bacterium can therefore be introduced in a fixed form for treatment of wastewater containing nitrate with a high C/N ratio.

Aerobiosis↗