Engineering topics

Lal, Ravi

Publications and source records attributed to Lal, Ravi.

3 records

DOE OSTI2020

A bimodular PKS platform that expands the biological design space

Traditionally engineered to produce novel bioactive molecules, Type I modular polyketide synthases (PKSs) could be engineered as a new biosynthetic platform for the production of de novo fuels, commodity chemicals, and specialty chemicals. Previously, our investigations manipulated the first module of the lipomycin PKS to produce short chain ketones, 3-hydroxy acids, and saturated, branched carboxylic acids. Building upon this work, we have expanded to multi-modular systems by engineering the first two modules of lipomycin to generate unnatural polyketides as potential biofuels and specialty chemicals in Streptomyces albus. First, we produce 20.6 mg/L of the ethyl ketone, 4,6 dimethylheptanone through a reductive loop exchange in LipPKS1 and a ketoreductase knockouts in LipPKS2. Furthermore, we then show that an AT swap in LipPKS1 and a reductive loop exchange in LipPKS2 can produce the potential fragrance 3-isopropyl-6-methyltetrahydropyranone. Highlighting the challenge of maintaining product fidelity, in both bimodular systems we observed side products from premature hydrolysis in the engineered first module and stalled dehydration in reductive loop exchanges. Collectively, our work expands the biological design space and moves the field closer to the production of "designer" biomolecules.

09 BIOMASS FUELS↗

DOE OSTI2020

Chemoinformatic-Guided Engineering of Polyketide Synthases

Polyketide synthase (PKS) engineering is an attractive method to generate new molecules such as commodity, fine and specialty chemicals. A significant challenge is re-engineering a partially reductive PKS module to produce a saturated β-carbon through a reductive loop (RL) exchange. In this work, we sought to establish that chemoinformatics, a field traditionally used in drug discovery, offers a viable strategy for RL exchanges. We first introduced a set of donor RLs of diverse genetic origin and chemical substrates into the first extension module of the lipomycin PKS (LipPKS1). Product titers of these engineered unimodular PKSs correlated with chemical structure similarity between the substrate of the donor RLs and recipient LipPKS1, reaching a titer of 165 mg/L of short-chain fatty acids produced by the host Streptomyces albus J1074. Finally, expanding this method to larger intermediates that require bimodular communication, we introduced RLs of divergent chemosimilarity into LipPKS2 and determined triketide lactone production. Collectively, we observed a statistically significant correlation between atom pair chemosimilarity and production, establishing a new chemoinformatic method that may aid in the engineering of PKSs to produce desired, unnatural products.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

NASA NTRS2007

Research Infrastructure Challenges for Graduate Programs in STEM Disciplines at Minority Institutions

It is much more challenging to perform experimental research functions at many minority institutions, because of lack of adequate research infrastructure. This is especially true if one wishes to initiate and implement masters and doctoral degree program in physics. In the present paper, an attempt is made to discuss the various hurdles encountered by the authors in the establishment of Master's and Doctoral degree programs in physics at one of the HBCUs (Historically Black Colleges and Universities). The department got no special or necessary treatment and faculty members are asked to teach as much course work as any other undergraduate department on the campus. It was very hard to convince university administration that giving less teaching load to research producing department faculty, shall culminate in abundant funding for the future years. This scenario created an extra heavy pressure on the faculty to continue the program. Some of the challenges included the resistance of some faculty and administrators to change, lack of sufficient release time for research producing faculty, and potential variation in funding or support with changes in the state education budget proration or members of the administration. In spite of the indirect cost recovery, very little infrastructure facilities was provided and the federal funding agencies did not want to interfere in the administration of the university. Various issues of recruiting and mentoring minority students, retention in the STEM disciplines as well as research infrastructure challenges at an HBCU university are presented.

Aggarwal, M. D.↗