Methods for producing designer esters and assessing alcohol acyltransferase specificity for ester biosynthesis
US12297480B2 · kind B2 · utility
Inventors
Key dates
| Filing date | Aug 26, 2022 |
| Grant date | May 13, 2025 |
| Priority date | — |
| Expiry date | Aug 26, 2042 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY02E50/10
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods of microbial screening for identifying alcohol acyltransferases for ester biosynthesis and submodules for ester pathways to produce butyryl-coenzyme A derived esters are disclosed. The method includes the introduction preselected plasmids into a respective host strain to form engineered microbes, in situ fermentation thereof followed by a colorimetric assay for quantification of production of the target ester. In situ fermentation includes inoculating each well of a microplate that have a culture media for producing target esters with one of the engineered microbes, adding an overlay of a solvent to each, and incubating the same. The colorimetric assay includes transfer of a quantity of the overlay from each well to respective clean wells of a new microplate, treatment of each well to form an iron-hydroxamic acid complex aqueous phase, centrifugation of the microplate, and measurement of the absorbance at 520 nm and comparison to a standard curve for the target ester.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.