Engineering topics

Larson, James

Publications and source records attributed to Larson, James.

6 records

DOE OSTI2025

Archaeal protein containing domain of unknown function 2193 undergoes oligomeric reconfiguration upon iron–sulfur cluster binding

Methanogenic archaea are particularly rich in iron–sulfur proteins, yet their roles remain largely enigmatic. Here, we characterized aMethanococcus voltae(Mvo) protein from the domain of unknown function (DUF) 2193 family, a group of proteins present primarily in archaea and characterized by a conserved cysteine‐rich C‐terminal motif.MvoDUF2193 was heterologously expressed and characterized by a range of spectroscopic and analytical methods. The results demonstrate thatMvoDUF2193 binds a single [4Fe–4S] cluster per subunit and that cluster occupancy regulates the transition from an apo tetramer to a [4Fe–4S] monomeric form. We hypothesize thatMvoDUF2193 serves a regulatory role in the cell, mediated by [Fe–S] cluster binding and changes in oligomeric state.

Biochemistry & Molecular Biology↗

DOE OSTI2024

Impact of mineral and non-mineral sources of iron and sulfur on the metalloproteome of Methanosarcina barkeri

Methanogens often inhabit sulfidic environments that favor the precipitation of transition metals such as iron (Fe) as metal sulfides, including mackinawite (FeS) and pyrite (FeS 2 ). These metal sulfides have historically been considered biologically unavailable. Nonetheless, methanogens are commonly cultivated with sulfide (HS - ) as a sulfur source, a condition that would be expected to favor metal precipitation and thus limit metal availability. Recent studies have shown that methanogens can access Fe and sulfur (S) from FeS and FeS 2 to sustain growth. As such, medium supplied with FeS 2 should lead to higher availability of transition metals when compared to medium supplied with HS - . Here, we examined how transition metal availability under sulfidic (i.e., cells provided with HS - as sole S source) versus non-sulfidic (cells provided with FeS 2 as sole S source) conditions impact the metalloproteome of Methanosarcina barkeri Fusaro. To achieve this, we employed size exclusion chromatography coupled with inductively coupled plasma mass spectrometry and shotgun proteomics. Significant changes were observed in the composition and abundance of iron, cobalt, nickel, zinc, and molybdenum proteins. Among the differences were alterations in the stoichiometry and abundance of multisubunit protein complexes involved in methanogenesis and electron transport chains. Furthermore, our data suggest that M. barkeri utilizes the minimal iron-sulfur cluster complex and canonical cysteine biosynthesis proteins when grown on FeS 2 but uses the canonical Suf pathway in conjunction with the tRNA-Sep cysteine pathway for iron-sulfur cluster and cysteine biosynthesis under sulfidic growth conditions.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Homologous acetone carboxylases select Fe(II) or Mn(II) as the catalytic cofactor

The Irving-Williams series refers to the predicted stabilities of transition metal complexes where the observed general stability for divalent first-row transition metal complexes increase across the row. Acetone carboxylases (ACs) use a coordinated divalent metal at their active site in the catalytic conversion of bicarbonate and acetone to form acetoacetate. Highly homologous ACs discriminate among different divalent metals at their active sites such that variations of the enzyme prefer Mn(II) over Fe(II), defying Irving-Williams-predicted behavior. Defining the determinants that promote metal discrimination within the first-row transition metals is of broad fundamental importance in understanding metal-mediated catalysis and metal catalyst design.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Biochemical and structural characterization of an aromatic ring–hydroxylating dioxygenase for terephthalic acid catabolism

Significance More than 400 million tons of plastic waste is produced each year, the overwhelming majority of which ends up in landfills. Bioconversion strategies aimed at plastics have emerged as important components of enabling a circular economy for synthetic plastics, especially those that exhibit chemically similar linkages to those found in nature, such as polyesters. The enzyme system described in this work is essential for mineralization of the xenobiotic components of poly(ethylene terephthalate) (PET) in the biosphere. Our description of its structure and substrate preferences lays the groundwork for in vivo or ex vivo engineering of this system for PET upcycling.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2021

Techno-Economic Analysis of decentralized preprocessing systems for fast pyrolysis biorefineries with blended feedstocks in the southeastern United States

This study evaluated the economic feasibility of fast pyrolysis biorefineries fed with blended pine residues and switchgrass in the Southeastern U.S. with different supply chain design. Previous techno-economic analyses (TEA) have focused on either blended biomass or decentralized preprocessing without investigating the impacts of varied process parameters, technology options, and real-world biomass distribution. This study fills the literature gap by modeling scenarios for different biomass blending ratios, biorefinery and preprocessing site (so-called depot) capacities, and alternative preprocessing technologies. High-resolution, real-world geospatial data were analyzed using Geographic Information Systems to facilitate supply chain design and TEA. For a decentralized system, the minimum fuel selling price (MFSP) of biofuel was 3.92–4.33 per gallon gasoline equivalent (GGE), while the MFSP for the centralized biorefinery at the same capacities ranged between 3.75–4.02/GGE. Implementing a high moisture pelleting process depot rather than a conventional pelleting process lowered the MFSP by 0.03–0.17/GGE. Scenario analysis indicated decreased MFSP with increasing biorefinery capacities but not necessarily with increasing depot size. Medium-size depots (500 OMDT/day) achieved the lowest MFSP. Here, this analysis identified the optimal blending ratios for two preprocessing technologies at varied depot sizes. Counterintuitively, increasing the proportion of higher cost switchgrass reduced the MFSP for large biorefineries (>5000 ODMT/day), but increased the MFSP for small biorefineries (1000–2500 ODMT/day). Although the decentralized systems have a higher MFSP based on current analysis, it has other potential benefits such as mitigated supply chain risks and improved feedstock quality that are difficult to be quantified in this TEA.

09 BIOMASS FUELS↗

DOE OSTI2020

Impacts of uncertain feedstock quality on the economic feasibility of fast pyrolysis biorefineries with blended feedstocks and decentralized preprocessing sites in the Southeastern United States

Abstract This study performs techno‐economic analysis and Monte Carlo simulations (MCS) to explore the effects that variations in biomass feedstock quality have on the economic feasibility of fast pyrolysis biorefineries using decentralized preprocessing sites (i.e., depots that produce pellets). Two biomass resources in the Southeastern United States, that is, pine residues and switchgrass, were examined as feedstocks. A scenario analysis was conducted for an array of different combinations, including different pellet ash control levels, feedstock blending ratios, different biorefinery capacities, and different biorefinery on‐stream capacities, followed by a comparison with the traditional centralized system. MCS results show that, with depot preprocessing, variations in the feedstock moisture and feedstock ash content can be significantly reduced compared with a traditional centralized system. For a biorefinery operating at 100% of its designed capacity, the minimum fuel selling price (MFSP) of the decentralized system is $3.97–$4.39 per gallon gasoline equivalent (GGE) based on the mean value across all scenarios, whereas the mean MFSP for the traditional centralized system was $3.79–$4.12/GGE. To understand the potential benefits of highly flowable pellets in decreasing biorefinery downtime due to feedstock handling and plugging problems, this study also compares the MFSP of the decentralized system at 90% of its designed capacity with a traditional system at 80%. The analysis illustrates that using low ash pellets mixed with switchgrass and pine residues generates a more competitive MFSP. Specifically, for a biorefinery designed for 2,000 oven dry metric ton per day, running a blended pellet made from 75% switchgrass and 25% pine residues with 2% ash level, and operating at 90% of designed capacity could make an MFSP between $4.49 and $4.71/GGE. In contrast, a traditional centralized biorefinery operating at 80% of designed capacity marks an MFSP between $4.72 and $5.28.

09 BIOMASS FUELS↗