Results for “Engine” — Page 51

NASA NTRS2016

Modeling of Highly Instrumented Honeywell Turbofan Engine Tested with Ice Crystal Ingestion in the NASA Propulsion System Laboratory

The Propulsion Systems Laboratory (PSL), an altitude test facility at NASA Glenn Research Center, has been used to test a highly instrumented turbine engine at simulated altitude operating conditions. This is a continuation of the PSL testing that successfully duplicated the icing events that were experienced in a previous engine (serial LF01) during flight through ice crystal clouds, which was the first turbofan engine tested in PSL. This second model of the ALF502R-5A serial number LF11 is a highly instrumented version of the previous engine. The PSL facility provides a continuous cloud of ice crystals with controlled characteristics of size and concentration, which are ingested by the engine during operation at simulated altitudes. Several of the previous operating points tested in the LF01 engine were duplicated to confirm repeatability in LF11. The instrumentation included video cameras to visually illustrate the accretion of ice in the low pressure compressor (LPC) exit guide vane region in order to confirm the ice accretion, which was suspected during the testing of the LF01. Traditional instrumentation included static pressure taps in the low pressure compressor inner and outer flow path walls, as well as total pressure and temperature rakes in the low pressure compressor region. The test data was utilized to determine the losses and blockages due to accretion in the exit guide vane region of the LPC. Multiple data points were analyzed with the Honeywell Customer Deck. A full engine roll back point was modeled with the Numerical Propulsion System Simulation (NPSS) code. The mean line compressor flow analysis code with ice crystal modeling was utilized to estimate the parameters that indicate the risk of accretion, as well as to estimate the degree of blockage and losses caused by accretion during a full engine roll back point. The analysis provided additional validation of the icing risk parameters within the LPC, as well as the creation of models for estimating the rates of blockage growth and losses.

Simulation↗

NASA NTRS2017

NASA Systems Engineering Handbook

The update of this handbook continues the methodology of the previous revision: a top-down compatibility with higher level Agency policy and a bottom-up infusion of guidance from the NASA practitioners in the field. This approach provides the opportunity to obtain best practices from across NASA and bridge the information to the established NASA systems engineering processes and to communicate principles of good practice as well as alternative approaches rather than specify a particular way to accomplish a task. The result embodied in this handbook is a top-level implementation approach on the practice of systems engineering unique to NASA. Material used for updating this handbook has been drawn from many sources, including NPRs, Center systems engineering handbooks and processes, other Agency best practices, and external systems engineering textbooks and guides. This handbook consists of six chapters: (1) an introduction, (2) a systems engineering fundamentals discussion, (3) the NASA program project life cycles, (4) systems engineering processes to get from a concept to a design, (5) systems engineering processes to get from a design to a final product, and (6) crosscutting management processes in systems engineering. The chapters are supplemented by appendices that provide outlines, examples, and further information to illustrate topics in the chapters. The handbook makes extensive use of boxes and figures to define, refine, illustrate, and extend concepts in the chapters.

ENGINEERING MANAGEMENT; HANDBOOKS; MANAGEMENT METH↗

NASA NTRSDate not supplied

Lessons Learned in Systems Engineering Availability and Recommendations for Mission Technical Leaders

In spaceflight missions at Goddard Space Flight Center (GSFC), the Mission System Engineer (MSE) is the technical leader of the overall engineering team and also is the Independent Engineering Technical Authority. The responsibility of this role includes the definition of the mission design architecture, concept of operations, and mission requirements, management of risk throughout the development, and verification and validation of the final system performance and function amongst other duties. This responsibility inherently requires time management, enabling focus on a balanced development with appropriate risk. Time is the most valuable resource of the MSE. The system engineer’s availability to interact with the development team (often product or component design leads and technicians, often in different worksites) to discover and mitigate mission risks during development is key to mission success. This paper presents examples from Lunar Reconnaissance Orbiter, Landsat 9, and Neutron star Interior Composition ExploreR (NICER) which represent in-house and out-of-house hardware builds. These examples demonstrate how interactions between the Mission Systems Engineers and other project and partner engineers result in discovery of critical risks, leading to early mitigation with significant cost and performance savings. These three missions would have suffered test failures or on-orbit failures had their MSEs not set aside time to visit engineers and technicians that were working on key pieces of space flight hardware. Availability is more than just time; it is openness to listen to concerns and questions. It begins by building a level of trust in the team that it is safe to ask questions or share concerns without the fear of blame or additional workload. It also requires enabling informal conversations (over lunch, coffee, in the clean room, or at the team members desk, etc.) where key information can be exchanged, and team members may even provide an easy-to-implement mitigation idea for another subsystem. Availability is a highly valuable commodity and completely non-obvious to protect and optimize. The natural tendency of engineers is to keep themselves busy with solving problems that they know about. This paper is encouraging MSEs to resist this tendency to try and solve all the complex problems themselves and actively devote daily time to learning and solving problems that are found with informal communications with other team members.

Lessons Learned↗

NASA NTRSDate not supplied

How to Educate Decision Makers on the Value and Necessity of Modal Testing and Model Correlation: Tips for Young Engineers

Engineers need to effectively communicate the justification and value of their modal testing and model correlation in terminology familiar to decision makers as it relates to the program’s risk tolerance. This communication must relate to the program’s risk tolerance and the metrics used to judge the performance of both the program and individual decision makers. The challenge is the terminologies familiar to engineers and decision makers are quite different and seemingly unrelated. The engineering profession has developed a specific terminology to solve highly technical issues, which are many times themselves unique to very specific engineering problems. It is all too easy for engineers to believe that everyone in their organization, including the decision makers, has an intrinsic understanding of what they do and the value it brings to the program’s success. This is especially true for young engineers who have recently spent the last four plus years in an academic engineering learning environment, which has a highly technical research oriented atmosphere. Effective communication with decision makers is increasingly important as the technical breadth and practical program and project experience level for up and coming decision makers diminishes. It is not unusual for the decision makers to have technical knowledge in a domain different from structural dynamics (e.g., electronics or systems). Competition among satellite manufactures has increased the focus on programmatic cost and ability to deliver on schedule. NASA programs are also seeing more restrictive programmatic cost and schedule constraints, which impact both analysis and testing. It should also be noted that a comprehensive suite of tests are required to verify a satellite’s design capability with some margin. These tests include static strength verification tests, shock, acoustic, and vibration tests (sine and random) of systems, subsystems, and components. Each of these verification tests provide opportunities for model correlation and risk reduction. It is important to recognize dynamic loads/modal test models may not include all of the flight hardware (i.e., harness, coax, waveguides, connectors, etc.) and the previously mentioned tests are still required for qualification/verification of the design. This paper provides tips to young engineers on how to bridge this communications gap, have a better understanding of the environment in which decision makers operate, and assist them to better support successful missions. While this paper primarily focuses on modal testing and model correlation as related to spacecraft missions, the concepts and recommendations presented here are equally applicable to other fields such as aeronautics, automotive, power generation, etc.

Decision Maker↗

NASA NTRSDate not supplied

How to Educate Decision Makers on the Value and Necessity of Modal Testing and Model Correlation: Tips for Young Engineers

Engineers need to effectively communicate the justification and value of their modal testing and model correlation in terminology familiar to decision makers as it relates to the program’s risk tolerance. This communication must relate to the program’s risk tolerance and the metrics used to judge the performance of both the program and individual decision makers. The challenge is the terminologies familiar to engineers and decision makers are quite different and seemingly unrelated. The engineering profession has developed a specific terminology to solve highly technical issues, which are many times themselves unique to very specific engineering problems. It is all too easy for engineers to believe that everyone in their organization, including the decision makers, has an intrinsic understanding of what they do and the value it brings to the program’s success. This is especially true for young engineers who have recently spent the last four plus years in an academic engineering learning environment, which has a highly technical research oriented atmosphere. Effective communication with decision makers is increasingly important as the technical breadth and practical program and project experience level for up and coming decision makers diminishes. It is not unusual for the decision makers to have technical knowledge in a domain different from structural dynamics (e.g., electronics or systems). Competition among satellite manufactures has increased the focus on programmatic cost and ability to deliver on schedule. NASA programs are also seeing more restrictive programmatic cost and schedule constraints, which impact both analysis and testing. It should also be noted that a comprehensive suite of tests are required to verify a satellite’s design capability with some margin. These tests include static strength verification tests, shock, acoustic, and vibration tests (sine and random) of systems, subsystems, and components. Each of these verification tests provide opportunities for model correlation and risk reduction. It is important to recognize dynamic loads/modal test models may not include all of the flight hardware (i.e., harness, coax, waveguides, connectors, etc.) and the previously mentioned tests are still required for qualification/verification of the design. This paper provides tips to young engineers on how to bridge this communications gap, have a better understanding of the environment in which decision makers operate, and assist them to better support successful missions. While this paper primarily focuses on modal testing and model correlation as related to spacecraft missions, the concepts and recommendations presented here are equally applicable to other fields such as aeronautics, automotive, power generation, etc.

Decision Maker↗

NASA NTRSDate not supplied

Transient Optimization of a Gas Turbine Engine

Gas turbine engines are the primary power plants for modern commercial aircraft. Transients prompted by significant changes in thrust or power demand are common and unavoidable. Extreme transient scenarios such as those associated with a go-around during a landing attempt are possible and must be accounted for in the design of the engine and its controller. Engine transients tend to cause a reduction in compressor operability margin, which must be addressed by the engine control system and accounted for in the engine design to prevent events such as compressor stall/surge and combustor blow out. Transient operability concerns typically lead to compromises in the engine design that sacrifice efficiency and/or limit responsiveness. Transient operability is typically managed by logic that limits the fuel flow command. If this logic is not optimized, then the potential for valuable performance could be lost. This study presents a strategy for optimizing the transient limit logic and proposes a strategy for updating the control logic over the lifespan of the engine. The results demonstrate significant improvements in transient operability. For example, of the results at sea level static conditions demonstrated a 31% reduction in the usage of the high pressure compressor operability stack during a snap acceleration transient. Furthermore, a reinforcement learning algorithm is demonstrated to modify the transient logic as the engine degrades to minimize response time while respecting a prescribed compressor operability margin limit. A simple demonstration of the reinforcement learning algorithm resulted in a thrust response time reduction of ~11.8%.

transient↗

NASA NTRSDate not supplied

Transient Optimization of a Gas Turbine Engine

Gas turbine engines are the primary power plants for modern commercial aircraft. Transients prompted by significant changes in thrust or power demand are common and unavoidable. Extreme transient scenarios such as those associated with a go-around during a landing attempt are possible and must be accounted for in the design of the engine and its controller. Engine transients tend to cause a reduction in compressor operability margin, which must be addressed by the engine control system and accounted for in the engine design to prevent events such as compressor stall/surge and combustor blow out. Transient operability concerns typically lead to compromises in the engine design that sacrifice efficiency and/or limit responsiveness. Transient operability is typically managed by logic that limits the fuel flow command. If this logic is not optimized, then the potential for valuable performance could be lost. This study presents a strategy for optimizing the transient limit logic and proposes a strategy for updating the control logic over the lifespan of the engine. The results demonstrate significant improvements in transient operability. For example, of the results at sea level static conditions demonstrated a 31% reduction in the usage of the high pressure compressor operability stack during a snap acceleration transient. Furthermore, a reinforcement learning algorithm is demonstrated to modify the transient logic as the engine degrades to minimize response time while respecting a prescribed compressor operability margin limit. A simple demonstration of the reinforcement learning algorithm resulted in a thrust response time reduction of ~11.8%.

transient↗

NASA NTRSDate not supplied

Transient Optimization of a Gas Turbine Engine

Gas turbine engines are the primary power plants for modern commercial aircraft. Transients prompted by significant changes in thrust or power demand are common and unavoidable. Extreme transient scenarios such as those associated with a go-around during a landing attempt are possible and must be accounted for in the design of the engine and its controller. Engine transients tend to cause a reduction in compressor operability margin, which must be addressed by the engine control system and accounted for in the engine design to prevent events such as compressor stall/surge and combustor blow out. Transient operability concerns typically lead to compromises in the engine design that sacrifice efficiency and/or limit responsiveness. Transient operability is typically managed by logic that limits the fuel flow command. If this logic is not optimized, then the potential for valuable performance could be lost. This study presents a strategy for optimizing the transient limit logic and proposes a strategy for updating the control logic over the lifespan of the engine. The results demonstrate significant improvements in transient operability. For example, of the results at sea level static conditions demonstrated a 31% reduction in the usage of the high pressure compressor operability stack during a snap acceleration transient. Furthermore, a reinforcement learning algorithm is demonstrated to modify the transient logic as the engine degrades to minimize response time while respecting a prescribed compressor operability margin limit. A simple demonstration of the reinforcement learning algorithm resulted in a thrust response time reduction of ~11.8%.

transient↗

DOE OSTI2018

Method of making an aero-derivative gas turbine engine

A method of making an aero-derivative gas turbine engine (100) is provided. A combustor outer casing (68) is removed from an existing aero gas turbine engine (60). An annular combustor (84) is removed from the existing aero gas turbine engine. A first row of turbine vanes (38) is removed from the existing aero gas turbine engine. A can annular combustor assembly (122) is installed within the existing aero gas turbine engine. The can annular combustor assembly is configured to accelerate and orient combustion gasses directly onto a first row of turbine blades of the existing aero gas turbine engine. A can annular combustor assembly outer casing (108) is installed to produce the aero-derivative gas turbine engine (100). The can annular combustor assembly is installed within an axial span (85) of the existing aero gas turbine engine vacated by the annular combustor and the first row of turbine vanes.

42 ENGINEERING↗

DOE OSTI2020

Development of a Physics-Based Combustion Model for Engine Knock Prediction

The objective of this project is to improve the prediction of engine knock by developing a new combustion modeling framework. Engine knock is a limiting factor to constrain the increase of fuel efficiency for spark ignition (SI) engines in most passenger cars. Efforts to increase fuel efficiency, increasing the compression ratio or downsizing, lead to the increase in the tendency of the knock occurrence. The knock is an undesired ignition of the end-gas, unburned fuel/air mixture ahead of the spark-ignited premixed flame, resulting in rapid in-cylinder pressure rises and engine damages. The combustion modeling framework developed in this project can consider turbulence-chemistry interactions during end-gas ignition, while using a reasonably detailed chemical mechanism developed for ignition and combustion reactions under engine relevant conditions, and the subtle characteristics of spark-ignited flame propagation. It is developed in the context of large eddy simulation (LES), which can capture stochastic in-cylinder processes. The developed model is incorporated into a commercial software for engine simulation, CONVERGE CFD, as a user defined function, and validated. Engine knock and knock-free experiments as well as direct numerical simulation (DNS) of end-gas ignition in homogeneous turbulence are performed to help model development and provide data sets for model validation. With further validation, the developed model is expected to advance the predictive capability for engine knock simulations and thus contribute to improving the fuel efficiency.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

DOE OSTI2019

The Effect of EGR on Knock Suppression, Efficiency, and Emissions in a Stoichiometric, Spark Ignited, Natural Gas Engine

Exhaust gas recirculation (EGR) is widely employed on internal combustion engines for NOx reduction and knock suppression. In this study a comprehensive analysis was performed to determine the effect of EGR on efficiency, emissions, and knock suppression of stoichiometric, spark ignited, natural gas engines. Tests were conducted using a single cylinder, variable compression ratio, cooperative fuel research (CFR) engine at 900 RPM, engine load of 800 kPa indicated mean effective pressure (IMEP), and stoichiometric conditions. The tests were performed using a custom EGR system capable of providing a range of EGR displacement rates from 0% to 40% of the intake charge at engine power levels between 1.5 and 3.5 kW. The experimental measurements included the variance of EGR rate, compression ratio, boost pressure, and location of 50% mass fraction burned (CA50). CA50 was controlled with ignition timing using a Woodward Large Engine Control Module. The use of EGR was found to expand the knock limit, while reducing NOx emissions and increasing engine efficiency as much as 4.5% through increased compression ratio and power density. The highest EGR rate that demonstrated beneficial gains on the CFR engine was 23%; the optimal EGR rate was found to be 19%.

Cooperative Fuel Research Engine, Exhaust Gas Reci↗

DOE OSTI2022

New Directions for Chemical Engineering

Over the past century, the work of chemical engineers has helped transform societies and the lives of individuals, from the synthetic fertilizers that helped feed the world to the development of novel materials used in fuels, electronics, medical devices, and other products. Chemical engineers' ability to apply systems-level thinking from molecular to manufacturing scales uniquely positions them to address today’s most pressing problems, including climate change and the overuse of resources by a growing population. New Directions for Chemical Engineering details a vision to guide chemical engineering research, innovation, and education over the next few decades. This report calls for new investments in U.S. chemical engineering and the interdisciplinary, cross-sector collaborations necessary to advance the societal goals of transitioning to a low-carbon energy system, ensuring our production and use of food and water is sustainable, developing medical advances and engineering solutions to health equity, and manufacturing with less waste and pollution. The report also calls for changes in chemical engineering education to ensure the next generation of chemical engineers is more diverse and equipped with the skills necessary to address the challenges ahead.

National Academies of Sciences, Engineering, and M↗

DOE OSTI2023

SPC-70486 Rev 0 Stirling Engine Prototype Performance Specification

Idaho National Laboratory (INL) is developing a small microreactor to produce electrical power utilizing a small nuclear core and Stirling engines under the Microreactor Application Research Validation and Evaluation (MARVEL) program. The purpose of this performance specification is to define requirements to build and test the Stirling Engine prototype, including interfacing with Kinnetech in the development/test of the engines. This specification includes prototyping components only and are not for use in the reactor. Prototype testing is a preliminary test to be conducted with conceptual equipment from which a final design will be developed. The results of the testing defined herein will not be used as a qualification test. The Stirling Engines have an engine controller module (QEC). Currently, the QEC doesn’t have a means to stop the engine. An analog controlled engine braking system must be developed and tested with the engine prototype. Any conflict between this specification and any supplementary specifications in the procurement documents requires written clarification from the Contractor prior to proceeding with any work. Any deviation from the procurement documents requires approval by the Contractor with the change request process.

21 - SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLAN↗

DOE OSTI2022

Investigation of Cycle-to-Cycle Variations in Internal Combustion Engine Using Proper Orthogonal Decomposition

The understanding, modeling and control of the cycle-to-cycle variation (CCV) in the modern internal combustion engine (ICE) is a key scientific challenge to achieve stable engine operation. High CCV in the engine combustion chamber may contribute to partial burn, misfire and knock, which adversely affects the engine performance and may potentially damage the engine. The objective of the current study is to leverage high-fidelity numerical simulations to improve the understanding of the causes of CCV. Using the massively parallel code, Nek5000, multi-cycle, wall-resolved large-eddy simulations (LES) were performed for the General Motors (GM), Transparent Combustion Chamber (TCC-III) optical engine under motored operating conditions. Further, the large-scale structures of the in-cylinder flow were investigated using a triple proper orthogonal decomposition (POD) technique to explore the characteristics of different parts of the flow and their contributions to CCV. The kinetic energy of the subset of flow structures were determined and correlated between the intake and compression strokes. The insights from the analysis of the large-scale flow structures will be used to assist the development of improved engine designs with reduced CCV and enhance the engine performance.

33 ADVANCED PROPULSION SYSTEMS↗

DOE OSTI2020

Effects of blending C3-C4 alcohols on motor gasoline properties and performance of spark ignition engines: A review

Supplementing petroleum fuels with sustainable and renewable alternatives is a good option for increasing the sustainability of transportation fuels. Alcohols are particularly attractive blendstocks for spark ignition engines, mainly due to their desirable fuel properties including high octane, evaporative cooling, and reduced sooting propensity. Although the use of SI engines is widespread around the world, predominately for light duty vehicles, concerns about CO2 emissions and other sustainability issues necessitate increased engine efficiencies, reduced tailpipe pollutants, and lower lifecycle carbon emissions. The intelligent blending of C2-C4 alcohols into motor gasoline is a viable method for achieving these goals. There are a multitude of ways to produce renewable alcohols such as through fermentation from first-generation feedstocks (sugar and corn) and second-generation feedstocks (lignocellulosic biomass), or by gasification and mixed alcohol fuel synthesis routes from lignocellulosic biomass. Currently ethanol is extensively used in motor gasoline fuels worldwide and although many have proposed the use of C3 and C4 alcohols in motor gasoline as an improvement over ethanol, the higher cost of production and lack of clear definition as to their benefit over ethanol when blended into motor gasoline have led to slow acceptance into the market. In this review, special emphasis is placed on the effects of blending C3 and C4 alcohols into motor gasoline in terms of physicochemical properties, volatility behavior, and engine performance when compared to ethanol blends. Furthermore, the impact of blending C3 and C4 alcohols with gasoline on emission (particulate matter, nitrogen oxides, carbon monoxide, hydrocarbons, and unregulated oxygenates) and combustion (volumetric efficiency, thermal efficiency, fuel consumption, and cold performance) characteristics is discussed. Although there are some disagreements in the literature over the effect of alcohols predominately around the type of SI engine, i.e., port fuel injected versus direct fuel injected and engine operating mode, generally it is stated that alcohols can potentially reduce soot, unburned hydrocarbons and CO emissions while increasing thermal efficiency when proper engine configuration/calibration is used. Lastly, research that must be conducted to find the optimum combination of alcohol blends and engine configurations is highlighted and discussed.

09 BIOMASS FUELS↗

DOE OSTI2021

Experimental validation of shock propagation through a foam with engineered macro-pores

The engineered macro-pore foam provides a new way to study thermonuclear burn physics by utilizing capsules containing deuterated (D) foam and filling tritium (T) gas in the engineered macro-pores. The implosion of a thermonuclear capsule filled with an engineered macro-pore foam will be complex due to the interaction of a shock wave with the engineered macro-pores. It is our goal to quantify how substantially complex foam structures affect the shape of shock and bulk shock speed. A cylinder-shape shock tube experiment has been designed and performed at the Omega Laser Facility. In order to examine how a foam structure will affect shock propagation, we performed several tests varying (1) engineered macro-pore size, (2) average foam density, and (3) with/without neopentane (C 5 H 12 ) gas. X-ray radiographic data indicate that shock speed through engineered macro-pore foams depends strongly on average foam density and less on pore size. In this work, experimental shock propagation data helped guide two numerical simulation approaches: (1) a 2D simulation with homogenizing foams rather than explicitly simulating engineered macro-pores and (2) a 2D toroidal-pore approximation adopting a toroidal-tube geometry to model engineered macro-pores.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

DOE OSTI2025

Comparison of a Full-Scale and a 1:10 Scale Low-Speed Two-Stroke Marine Engine Using Computational Fluid Dynamics

International marine shipping is a growing component of international trade; a vast majority of all the world’s goods are being transported on large ocean-going vessels. The International Maritime Organization (IMO) introduced the Energy Efficiency Design Index in 2013, a regulatory framework of associated metrics for reducing emissions of CO 2 per tonne-mile from shipping by approximately 10% each decade. Therefore, decarbonizing the maritime sector requires the development of new fuel sources. Because of the extremely large physical size of the internal combustion engines present in shipping vessels, experimental iterative development of the engine and fuel system is cost-prohibitive. Thus, the ability to perform combustion system development in a scaled platform that can be more easily operated and modeled computationally is of interest. To that end, scaling relationships are needed to translate the results from a smaller engine to a larger counterpart. Scaling studies to date have been restricted to low scaling ratios, four-stroke light-duty engines, and under-resolved computational fluid dynamic simulations that likely do not accurately capture the physics of scaling. In this work, computational models of a 1:10 scale and a full-scale two-stroke crosshead low-speed marine engine were created and validated against experiments obtained in a real 1:10 scale engine installed at Oak Ridge National Laboratory. Further, due to the large size of the full-scale engine, the model required large high-performance computing resources to be evaluated. The availability of high-performance computing resources at the Department of Energy’s Leadership Computing Facilities is an enabler of the current work. The results of the small- and large-scale engine simulations were compared to analyze the effectiveness of the appropriate scaling laws under these extreme scaling ratio conditions.

33 ADVANCED PROPULSION SYSTEMS↗

DOE OSTI2024

Tools for genetic engineering and gene expression control in Novosphingobium aromaticivorans and Rhodobacter sphaeroides

ABSTRACT Alphaproteobacteria have a variety of cellular and metabolic features that provide important insights into biological systems and enable biotechnologies. For example, some species are capable of converting plant biomass into valuable biofuels and bioproducts that have the potential to contribute to the sustainable bioeconomy. Among the Alphaproteobacteria, Novosphingobium aromaticivorans , Rhodobacter sphaeroides , and Zymomonas mobilis show promise as organisms that can be engineered to convert extracted plant lignin or sugars into bioproducts and biofuels. Genetic manipulation of these bacteria is needed to introduce engineered pathways and modulate expression of native genes with the goal of enhancing bioproduct output. Although recent work has expanded the genetic toolkit for Z. mobilis , N. aromaticivorans and R. sphaeroides still need facile, reliable approaches to deliver genetic payloads to the genome and to control gene expression. Here, we expand the platform of genetic tools for N. aromaticivorans and R. sphaeroides to address these issues. We demonstrate that Tn 7 transposition is an effective approach for introducing engineered DNA into the chromosome of N. aromaticivorans and R. sphaeroides . We screen a synthetic promoter library to identify isopropyl β-D-1-thiogalactopyranoside-inducible promoters with regulated activity in both organisms (up to ~15-fold induction in N. aromaticivorans and ~5-fold induction in R. sphaeroides ). Combining Tn 7 integration with promoters from our library, we establish CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) interference systems for N. aromaticivorans and R. sphaeroides (up to ~10-fold knockdown in N. aromaticivorans and R. sphaeroides ) that can target essential genes and modulate engineered pathways. We anticipate that these systems will greatly facilitate both genetic engineering and gene function discovery efforts in these species and other Alphaproteobacteria. IMPORTANCE It is important to increase our understanding of the microbial world to improve health, agriculture, the environment, and biotechnology. For example, building a sustainable bioeconomy depends on the efficient conversion of plant material to valuable biofuels and bioproducts by microbes. One limitation in this conversion process is that microbes with otherwise promising properties for conversion are challenging to genetically engineer. Here we report genetic tools for Novosphingobium aromaticivorans and Rhodobacter sphaeroides that add to the burgeoning set of tools available for genome engineering and gene expression in Alphaproteobacteria. Our approaches allow straightforward insertion of engineered pathways into the N. aromaticivorans or R. sphaeroides genome and control of gene expression by inducing genes with synthetic promoters or repressing genes using CRISPR interference. These tools can be used in future work to gain additional insight into these and other Alphaproteobacteria and to aid in optimizing yield of biofuels and bioproducts.

Hall, Ashley N.↗