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At least 163 records · Page 9
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

Engineering Pseudomonas putida for production of 3-hydroxyacids using hybrid type I polyketide synthases

Engineered type I polyketide synthases (T1PKSs) are a potentially transformative platform for the biosynthesis of small molecules. Due to their modular nature, T1PKSs can be rationally designed to produce a wide range of bulk or specialty chemicals. While heterologous PKS expression is best studied in microbes of the genus Streptomyces, recent studies have focused on the exploration of non-native PKS hosts. The biotechnological production of chemicals in fast growing and industrial relevant hosts has numerous economic and logistic advantages. With its native ability to utilize alternative feedstocks, Pseudomonas putida has emerged as a promising workhorse for the sustainable production of small molecules. Here, we outline the assessment of P. putida as a host for the expression of engineered T1PKSs and production of 3-hydroxyacids. After establishing the functional expression of an engineered T1PKS, we successfully expanded and increased the pool of available acyl-CoAs needed for the synthesis of polyketides using transposon sequencing and protein degradation tagging. This work demonstrates the potential of T1PKSs in P. putida as a production platform for the sustainable biosynthesis of unnatural polyketides.

Schmidt, Matthias↗
DOE OSTI2024

Structural basis for intermodular communication in assembly-line polyketide biosynthesis

Assembly-line polyketide synthases (PKSs) are modular multi-enzyme systems with considerable potential for genetic reprogramming. Understanding how they selectively transport biosynthetic intermediates along a defined sequence of active sites could be harnessed to rationally alter PKS product structures. Here, to investigate functional interactions between PKS catalytic and substrate acyl carrier protein (ACP) domains, we employed a bifunctional reagent to crosslink transient domain–domain interfaces of a prototypical assembly line, the 6-deoxyerythronolide B synthase, and resolved their structures by single-particle cryogenic electron microscopy (cryo-EM). Together with statistical per-particle image analysis of cryo-EM data, we uncovered interactions between ketosynthase (KS) and ACP domains that discriminate between intra-modular and inter-modular communication while reinforcing the relevance of conformational asymmetry during the catalytic cycle. Our findings provide a foundation for the structure-based design of hybrid PKSs comprising biosynthetic modules from different naturally occurring assembly lines.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Structural basis of the amidase ClbL central to the biosynthesis of the genotoxin colibactin

Colibactin is a genotoxic natural product produced by select commensal bacteria in the human gut microbiota. The compound is a bis-electrophile that is predicted to form interstrand DNA cross-links in target cells, leading to double-strand DNA breaks. The biosynthesis of colibactin is carried out by a mixed NRPS–PKS assembly line with several noncanonical features. An amidase, ClbL, plays a key role in the pathway, catalyzing the final step in the formation of the pseudodimeric scaffold. ClbL couples α-aminoketone and β-ketothioester intermediates attached to separate carrier domains on the NRPS–PKS assembly. Here, the 1.9 Å resolution structure of ClbL is reported, providing a structural basis for this key step in the colibactin biosynthetic pathway. The structure reveals an open hydrophobic active site surrounded by flexible loops, and comparison with homologous amidases supports its unusual function and predicts macromolecular interactions with pathway carrier-protein substrates. Modeling protein–protein interactions supports a predicted molecular basis for enzyme–carrier domain interactions. Overall, the work provides structural insight into this unique enzyme that is central to the biosynthesis of colibactin.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Variability and Spectral Characteristics of Three Flaring Gamma-Ray Quasars Observed by VERITAS and Fermi-LAT

Flat-spectrum radio quasars (FSRQs) are the most luminous blazars at GeV energies but only rarely emit detectable fluxes of TeV gamma rays, typically during bright GeV flares. We explore the gamma-ray variability and spectral characteristics of three FSRQs that have been observed at GeV and TeV energies by Fermi-LAT and VERITAS, making use of almost 100 hr of VERITAS observations spread over 10 yr: 3C 279, PKS 1222+216, and Ton 599. We explain the GeV flux distributions of the sources in terms of a model derived from a stochastic differential equation describing fluctuations in the magnetic field in the accretion disk and estimate the timescales of magnetic flux accumulation and stochastic instabilities in their accretion disks. We identify distinct flares using a procedure based on Bayesian blocks and analyze their daily and subdaily variability and gamma-ray energy spectra. Using observations from VERITAS, as well as Fermi, Swift, and the Steward Observatory, we model the broadband spectral energy distributions of PKS 1222+216 and Ton 599 during very high energy (VHE)–detected flares in 2014 and 2017, respectively, strongly constraining the jet Doppler factors and gamma-ray emission region locations during these events. Finally, we place theoretical constraints on the potential production of PeV-scale neutrinos during these VHE flares.

79 ASTRONOMY AND ASTROPHYSICS↗
NASA NTRS1973

Some aspects of cosmic synchrotron sources

Synchrotron emission is considered from individual particles which have small pitch angles and the general properties of synchrotron sources which mainly contain such particles, as well as the emissivities and degrees of circular polarization for specific source distributions. The limitation of synchrotron source models for optical pulsars and compact extragalactic objects are discussed, and it is shown that several existing models for the pulsar NP 0532 are inconsistent with the measured time variations and polarizations of the optical emission. Discussion is made also of whether the low frequency falloffs in the extragalactic objects PKS 2134 + 004, OQ 208, and NGC 1068 is due to emission from particles with small pitch angles or absorption by a thermal plasma or synchrotron self-absorption. It is concluded that the absorption interpretations cannot account for the turnover in the spectrum of PKS 2134 + 004. Measurements of polarization, angular structure, and X-ray flux are also described.

Epstein, R. I.↗
NASA NTRS1977

A spectroscopic study of selected quasars with absorption redshifts greater than emission redshifts

Spectroscopic data are presented for seven quasars having absorption systems whose redshift is greater than that of emission lines. The objects examined include PKS 1229-02, 3C 298, 3C 205, B194, PHL 1222, 0736-06, and PKS 0119-04. The redshift systems of each quasar are described in detail, the empirical distribution of the relative numbers of absorption systems vs. emission-absorption redshift difference is determined, and errors in this distribution due to selection of the sample and to the incompleteness and inhomogeneity of the data are discussed qualitatively. Six groups of possibilities are considered for the origin of absorption systems with redshifts higher than the emission redshifts. It is concluded that two separate mechanisms must be invoked: intervening clouds bound in a cluster of galaxies where the quasar is embedded (for systems with an absolute relative velocity not exceeding about 3000 km/s) and gravitational acceleration of material ejected by the quasar (for systems with a relative velocity of at least 3000 km/s).

Weymann, R. J.↗
NASA NTRS1978

X-ray observations of BL Lacertae objects

Results are reported for OSO 8 and HEAO 1 X-ray observations of the BL Lac objects Mrk 421, Mrk 501, and PKS 0548-322 over the energy range from 2 to 60 keV. It is found that the X-ray spectra of Mrk 421 and 501 are similar, exhibit statistically significant soft X-ray excesses, lack low-energy absorption, and are best fitted by power laws over the 2-30-keV range with respective photon indices of approximately 0.91 and 1.2 and respective hydrogen column densities of less than 8 x 10 to the 21st per sq cm and no more than 1.8 x 10 to the 22nd per sq cm. It is noted that the spectral index for both sources is identical, within errors, to the HF radio spectral index and that Mrk 421 experienced a factor of 6 decrease in the 2-10-keV band over a six-month period. A tentative identification of PKS 0548-322 with the source H0548-32 is suggested.

Mushotzky, R. F.↗
NASA NTRS1978

Optical spectroscopy of BL Lacertae objects and their relatives

Absorption lines from intervening gas along the line of sight to BL Lac objects are examined. The absorption line systems observed in several such objects are discussed, and detailed attention is given to the objects PKS 0735+178 and AO 0235+164. Random observations relevant to the nature of BL Lac objects are reviewed, including observations of steep-spectrum 3C QSOs and the peculiar object PKS 0528-250. The nature of the absorbing material is considered.

Smith, H. E.↗
NASA NTRS1979

X-ray emitting BL Lacertae objects located by the scanning modulation collimator experiment on HEAO 1

It is established that BL Lacertae objects constitute a class of X-ray emitter by precisely locating X-ray sources identified with Mrk 421, PKS 0548-322, PKS 2155-304, and 2A 1219+305. Irregular variations of the order of 1 mag, characteristic of BL Lacertae objects, are reported for the last two objects. The measurement of an extremely steep X-ray spectrum from Mrk 421, in contrast to previous flat X-ray spectra above 3 keV, suggests that different structural states of an accretion disk are being observed.

Schwartz, D. A.↗
NASA NTRS1979

Discovery of X-ray emission from two southern supernova remnants

Two soft X-ray sources positionally coincident with the supernova remnants PKS 1209-52 and RCW 103 have been discovered by using the A-2 experiment on HEAO 1. Their measured fluxes are, respectively, about 1.4 x 10 to the -10th erg/cm-sec (0.2-1.0 keV) and about 1.8 x 10 to the -10th erg/cm-sec (0.6-2.0 keV). Spectral data are used to derive physical parameters for each remnant. For PKS 1209-52 the parameters are suggestive of the remnant's being in an advanced evolutionary phase, with shock-heated interstellar material producing the soft X-ray emission. RCW 103, in contrast, is known from radio and optical data to be in an earlier evolutionary phase, and the soft X-ray flux is most likely due to emission originating in a reflected shock wave or in plasma evaporated from shock-heated interstellar clouds.

Tuohy, I. R.↗
NASA NTRS1982

Radio astronomy

Use of the Tidbinbilla Interferometer to refine the source positions in the Parkes 2.7 GHz survey of the southern sky is described. A result of the first phase of this work was the identification of a quasi-stellar object which appears to be the most remote object yet observed. This object has a red shift of 3.78 (PKS 2000-330, and a velocity of recession equal to 91% of that light. Based on Hubble's law, PKS 2000-330 appears to be 12 billion light years away.

Shaffer, R. D.↗
NASA NTRS1984

Variability of the ultraviolet spectrum of BL Lacertae objects

Attention is given to repeated observations of the two nearby BL Lac objects, Mrk 421 and PKS 0521-36, comparing the temporal behavior of their continuum in the 1300-3000 A range with that observed in intrinsically bright BL Lac objects and in Seyfert galaxies. Also discussed are the emission lines in the UV and optical spectrum of PKS 0521-36. It is suggested that the difference in variability pattern between BL Lac objects and Seyfert galaxies may be due to differences in the properties of the nonthermal continuum related to differences in accretion regimes, although it is more probably due to the presence of different continuum components in Seyfert galaxies. In BL Lac objects, the UV spectrum is likely to be synchrotron radiation from an optically thin source.

Ulrich, M. H.↗
NASA NTRS1987

EX56a study of extended X-ray emission around isolated galaxies EX56b identification and spectra of bright X-ray sources at high galactic latitude

The EXOSAT observations confirmed the identification and extended nature of PKS 2345-35. It gave a good 2 to 10 keV X-ray spectrum and a detailed spatial profile indicating asymmetry of the structure. In the high galactic latitidue investigation, the BL Lac object identified with the HEAO-1 source 1430+423 was detected, and the first X-ray spectrum was obtained. Several simulataneous observations of H0323+022 were obtained over a broad range of electromagnetic spectrum. Studies of luminous active galactic nuclei have given significant information on the spectrum of the quasar PKS 0558-504. In a study of Southern sky cataclysmic variables, the EXOSAT was used to determine the X-ray spectrum and search for periodicities in two objects. Studies of complete identifications have revealed that X-ray sources in two high galactic latitude fields are stars, and therefore are to be excluded from the Piccinotti extragalactic sample. Only one Piccinotti source remains to be identified.

Schwartz, Daniel A.↗
NASA NTRS1988

Upper limits from hard X-ray observations of five BL Lacertae objects

Results are presented from hard X-ray observations of the five brightest X-ray BL Lacertae objects: PKS 0548-322, Mrk 421 (=1101+384), 2A 1219+305, Mrk 501 (=1652+398), and PKS 2155-304. The observations covered the energy range 15-165 keV from August 1977 to December 1978. The results are compared with previous studies.

Bezler, M.↗
NASA NTRS1992

The origin of the optical emission lines associated with extragalactic radio sources

The observed extended emission line regions (EELRs) associated with radio sources are investigated with the objective of determining the characteristics of the ionization mechanisms and the possible effect of star formation. The sources included in the sample are 3C 227, 3C 277.3, 3C 305, Cen A (NGC 5128), NGC 7385, PKS 0349-27, PKS 2152-69, and Minkowski's Object. It is shown that the emission-line ratios of the EELRs considered can be explained by models which account for the coupled effect of photoionization and shock acting at different degrees. It is also shown that the EELR ionization is not due to young stars. The main energy sources of the EELR are identified.

Viegas, S. M.↗
NASA NTRS2002

The Relationship Between X-Rays and Relativistic Jets

We present recent multiwaveband observations centered on X-ray monitoring of blazars and the radio galaxy 3C 120 with the RXTE satellite, In 3C 120, we observed four X-ray dips, each followed by ejections of superluminal radio knots down the jet. This behavior, similar to that of the microquasar GRS 1915+105, is interpreted as infall of a piece of the inner accretion disk causing ejection of energy into the relativistic jet. The X-ray emission from the quasars PKS 1510-089, 3C 279, and 3C 273 is highly variable on timescales as short as approximately 1 day. Over 2 years, X-ray flares in PKS 1510-089 occurred about 2 weeks after radio outbursts, which can be explained by light-travel delays. In 3C 279 the X-ray and optical variations are usually well correlated, with very little, if any, time delay. We conclude that the X-ray and optical emission from blazars occurs near the radio core rather than close to the black hole.

Marscher, A. P.↗
NASA NTRS2002

Optical Microvariability of Blazars

We present the results of optical differential photometry of five blazers [PKS0219+428 (3C66A), PKS 0235+164 (AO 0235+16), H0414+019, PKS 0851+202 (OJ 187) and QSO 1807+698 (3C 371)] that were observed on 7 nights between November 05, 1997 and December 29, 1998, using the B and the V band filters. We have detected microvariations in four blazers (3C66A, AO 0235+16, H0414+019, and OJ 287). In addition, the light curve of AO 0235+16 has displayed a mini-flare when the brightness of this source was decreasing. Night-to-night variations have also been detected in 3C66A, H0414+019, and OJ 287. The results of our observations are discussed in the framework of accretion disk phenomena (magnetic flares or hot spots in accretion disks) and jet phenomena (plasma instabilities in jets).

Ghosh, K. K.↗
NASA NTRS2005

Probing the Relativistic Jets of Active Galactic Nuclei with Multiwavelength Monitoring

The work completed includes the analysis of observations obtained during Cycle 7 (March 2002-February 2003) of the Rossi X-ray Timing Explorer (RXTE). The project was part of a longer-term, continuing program to study the X-ray emission process in blazars and radio galaxies in collaboration with Dr. Ian McHardy (U. of Southampton, UK) and Prof. Thomas Balonek (Colgate U.). The goals of the program are to study the X-ray emission mechanism in blazars and radio galaxies and the relation of the X-ray emission to changes in the relativistic jet. The program includes contemporaneous brightness and linear polarization monitoring at radio and optical wavelengths, total and polarized intensity imaging at at 43 GHz with a resolution of 0.1 milliarcseconds with the VLBA, and well-sampled X-ray light curves obtained from a series of approved RXTE programs. The objects studied in the time period covered by the grant were 3C 120, 3C 279, PKS 1510-089, and 3C 273, all with radio jets containing bright knots that appear to move at superluminal speeds. During RXTE Cycle 7, the project was awarded RXTE time to monitor PKS 1510-089 two times per week, 3C 273 and 3C 279 three times per week, and 3C 120 four times per week. In addition, 3C273 and 3C 279 were observed several times per day during a ten-day period in April 2002. The X-ray data, including those from earlier cycles, were compared with radio measurements obtained in the centimeter-wave band by the monitoring program of Drs. Margo and Hugh Aller at the University of Michigan Radio Astronomy Observatory, monthly imaging observations with the VLBA at 43 GHz, and optical observations obtained at several telescopes around the world.

Marscher, Alan P.↗