Engineering topics

Tejera‐Nieves, Mauricio

Publications and source records attributed to Tejera‐Nieves, Mauricio.

2 records

DOE OSTI2024

Increasing thermostability of the key photorespiratory enzyme glycerate 3‐kinase by structure‐based recombination

As global temperatures rise, improving crop yields will require enhancing the thermotolerance of crops. One approach for improving thermotolerance is using bioengineering to increase the thermostability of enzymes catalysing essential biological processes. Photorespiration is an essential recycling process in plants that is integral to photosynthesis and crop growth. The enzymes of photorespiration are targets for enhancing plant thermotolerance as this pathway limits carbon fixation at elevated temperatures. We explored the effects of temperature on the activity of the photorespiratory enzyme glycerate kinase (GLYK) from various organisms and the homologue from the thermophilic alga Cyanidioschyzon merolae was more thermotolerant than those from mesophilic plants, including Arabidopsis thaliana. To understand enzyme features underlying the thermotolerance of C. merolae GLYK (CmGLYK), we performed molecular dynamics simulations using AlphaFold-predicted structures, which revealed greater movement of loop regions of mesophilic plant GLYKs at higher temperatures compared to CmGLYK. Based on these simulations, hybrid proteins were produced and analysed. These hybrid enzymes contained loop regions from CmGLYK replacing the most mobile corresponding loops of AtGLYK. Two of these hybrid enzymes had enhanced thermostability, with melting temperatures increased by 6 °C. One hybrid with three grafted loops maintained higher activity at elevated temperatures. Whilst this hybrid enzyme exhibited enhanced thermostability and a similar Km for ATP compared to AtGLYK, its Km for glycerate increased threefold. This study demonstrates that molecular dynamics simulation-guided structure-based recombination offers a promising strategy for enhancing the thermostability of other plant enzymes with possible application to increasing the thermotolerance of plants under warming climates.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

A 30% reduction in switchgrass rhizome reserves did not decrease biomass yield

Abstract A long‐standing question in perennial grass breeding and physiology is whether yield improvement strategies could compromise winter survival. Since perennial grasses rely on stored carbohydrates for winter maintenance and spring regrowth, yield improvement strategies could reduce winter survival if they increase biomass and grain yields at the expense of carbon allocation to storage. Therefore, it is crucial to comprehend the dependence of regrowth on storage reserves. We experimentally depleted switchgrass ( Panicum virgatum L.) rhizome reserves by storing rhizomes for 2 weeks at 5°C (control treatment) and 25°C (reserve‐depleted treatment). During the storage period rhizome respiration was 5.3× higher at 25°C (0.010 μmol CO 2 g −1 min −1 at 5°C vs. 0.054 μmol CO 2 g −1 min −1 at 25°C; p < 0.0001) and the starch content was depleted by 30% by the end of storage. Surprisingly, reserve‐depleted switchgrass had 60% larger leaf area (LA; LA control = 149 cm 2 pot −1 vs. LA depleted = 239 cm 2 pot −1 ; p = 0.013) and produced ~40% more aboveground biomass than control plants (9.46 g pot −1 vs. 6.63 g pot −1 ; p = 0.112). In addition, reserve‐depleted switchgrass restored its rhizome starch reserves to pre‐storage levels. Switchgrass showed a large plasticity among its source‐sink components to buffer the imposed reserve depletion. It increased plant photosynthesis by increasing the photosynthetic leaf area while keeping photosynthesis constant on a leaf area basis and readjusted the timing and activity of sink organs. These results suggest that switchgrass, and potentially other perennial grasses, largely over‐invest in storage reserves. Therefore, current breeding strategies in perennial grasses aimed to extend the aboveground growing season should not compromise crop persistence. Our study also has implications on long‐term yield dynamics as it highlights sink limitations as potential driver of the yield decline commonly observed in perennial grasses 5+ years after cultivation.

Tejera‐Nieves, Mauricio↗