Location: Bioenergy Research
Project Number: 5010-41000-189-008-I
Project Type: Interagency Reimbursable Agreement
Start Date: Sep 1, 2021
End Date: Aug 31, 2024
Objective:
The over-arching goal is to genetically engineer Bacillus coagulans for efficient conversion of cellulosic derived sugars into medium chain esters, which are marketed as industrial chemicals, specifically to:
Task 1: Rewire the endogenous metabolism of B. coagulans for efficient utilization of mixed sugars in lignocellulosic biomass hydrolysates
Task 2: Engineer the compatibility of thermostable enzymes and metabolic pathways for designer ester biosynthesis in B. coagulans.
Task 3: Enable biosystems design of B. coagulans for efficient conversion of lignocellulosic biomass to designer bioesters in mono- and co-cultures.
Approach:
We will work on extracting sugars from switchgrass and evaluating the native and engineered bacterial strains for growth and production on hydrolysate sugars.
1. Prepare concentrated sugars from switchgrass using pretreatment and enzymatic hydrolysis
2. Evaluate native and engineered B. coagulans for growth on refined sugar mixtures and unrefined switchgrass sugars (e.g. hydrolysate) at various concentrations
3. Optimize culture conditions to minimize lag phases and maximize product rates and yields
4. Apply adaptive laboratory evolution (ALE) using serial batch cultures to improve robustness for growth on unrefined switchgrass sugars as needed.
