Implications of Biorefinery Policy Incentives and Location-Specific Economic Parameters for the Financial Viability of Biofuels.

Cellulosic biofuels are part of a portfolio of solutions to address climate change; however, their production remains expensive and federal policy interventions (e.g., Renewable Fuel Standard) have not spurred broad construction of cellulosic biorefineries. A range of state-level interventions have also been enacted, but their implications for the financial viability of biorefineries are not well understood. To address this gap, this study evaluated the efficacy of 20 state-level tax incentives from 14 states and their interactions with other location-specific economic parameters (e.g., state income tax rates, electricity prices). To characterize implications of location-specific policies and parameters on biorefinery cash flows, we developed a new BioSTEAM Location-Specific Evaluation (BLocS) module for the open-source software BioSTEAM. Leveraging BLocS and BioSTEAM, we characterized the minimum ethanol selling price (MESP) for a cellulosic biorefinery (using corn stover as feedstock) and two conventional biorefineries (using corn or sugarcane as feedstock) for comparison. Among state-specific scenarios, nonincentivized MESPs for the corn stover biorefinery ranged from 0.74 $·L-1 (4.20 $·gallon gasoline equivalent [gge]-1) [0.69-0.79 $·L-1; 3.91-4.48 $·gge-1; Oklahoma] to 1.02 $·L-1 (5.78 $·gge-1) [0.95-1.09 $·L-1; 5.39-6.18 $·gge-1; New York], while the tax incentive-induced MESP reduction ranged from negligible (Virginia) to 5.78% [5.43-6.20%; Iowa]. Ultimately, this work can inform the design of policy incentives for biorefineries under specific deployment contexts.

Evaluation of 1,2-diacyl-3-acetyl triacylglycerol production in Yarrowia lipolytica.

Plants produce many high-value oleochemical molecules. While oil-crop agriculture is performed at industrial scales, suitable land is not available to meet global oleochemical demand. Worse, establishing new oil-crop farms often comes with the environmental cost of tropical deforestation. The field of metabolic engineering offers tools to transplant oleochemical metabolism into tractable hosts while simultaneously providing access to molecules produced by non-agricultural plants. Here, we evaluate strategies for rewiring metabolism in the oleaginous yeast Yarrowia lipolytica to synthesize a foreign lipid, 3-acetyl-1,2-diacyl-sn-glycerol (acTAG). Oils made up of acTAG have a reduced viscosity and melting point relative to traditional triacylglycerol oils making them attractive as low-grade diesels, lubricants, and emulsifiers. This manuscript describes a metabolic engineering study that established acTAG production at g/L scale, exploration of the impact of lipid bodies on acTAG titer, and a techno-economic analysis that establishes the performance benchmarks required for microbial acTAG production to be economically feasible.

Increasing metabolic pathway flux by using machine learning models.

Machine learning is transforming many industries through self-improving models that are fueled by big data and high computing power. The field of metabolic engineering, which uses cellular biochemical network to manufacture useful small molecules, has also witnessed the first wave of machine learning applications in the past five years, covering reaction route design, enzyme selection, pathway engineering and process optimization. This review focuses on pathway engineering, and uses a few recent studies to illustrate (1) how machine learning models can be useful in overcoming an evident rate-limiting step, and (2) how the models may be used to exhaustively search - or guide optimization algorithms to search - a large design space when the cellular regulation of the reaction network is more convoluted.

Mouse model of human RPE65 P25L hypomorph resembles wild type under normal light rearing but is fully resistant to acute light damage.

Human RPE65 mutations cause a spectrum of blinding retinal dystrophies from severe early-onset disease to milder manifestations. The RPE65 P25L missense mutation, though having <10% of wild-type (WT) activity, causes relatively mild retinal degeneration. To better understand these mild forms of RPE65-related retinal degeneration, and their effect on cone photoreceptor survival, we generated an Rpe65/P25L knock-in (KI/KI) mouse model. We found that, when subject to the low-light regime (∼100 lux) of regular mouse housing, homozygous Rpe65/P25L KI/KI mice are morphologically and functionally very similar to WT siblings. While mutant protein expression is decreased by over 80%, KI/KI mice retinae retain comparable 11-cis-retinal levels with WT. Consistently, the scotopic and photopic electroretinographic (ERG) responses to single-flash stimuli also show no difference between KI/KI and WT mice. However, the recovery of a-wave response following moderate visual pigment bleach is delayed in KI/KI mice. Importantly, KI/KI mice show significantly increased resistance to high-intensity (20 000 lux for 30 min) light-induced retinal damage (LIRD) as compared with WT, indicating impaired rhodopsin regeneration in KI/KI. Taken together, the Rpe65/P25L mutant produces sufficient chromophore under normal conditions to keep opsins replete and thus manifests a minimal phenotype. Only when exposed to intensive light is this hypomorphic mutation manifested physiologically, as its reduced expression and catalytic activity protects against the successive cycles of opsin regeneration underlying LIRD. These data also help define minimal requirements of chromophore for photoreceptor survival in vivo and may be useful in assessing a beneficial therapeutic dose for RPE65 gene therapy in humans.