Title: Data underlying chapter 4 Mechanistic insights in Lactobacillus brevis alcohol dehydrogenase: stability and active site role of Ser143 and Tyr156 

Authors: E.P.J. Jongkind, W.J. Kools, M. Pareek, S.C.L. Kamerlin, C.E. Paul

Link to publication: 10.3389/fctls.2023.1105948

Short description: This dataset contains data collected during experiments at Delft University of Technology, as Chapter 3 of the dissertation written by Ewald Jongkind

Contact person: Caroline Paul, c.e.paul@tudelft.nl
Delft University of Technology – Faculty of Applied Sciences
Van der Maasweg 9 , 2629 HZ, Delft, The Netherlands

***Purpose of the test campaign***
The purpose of these experiments was to investigate if with protein engineering, the alcohol dehydrogenase from Lactobacillus brevis, can catalyze reductive amination
The data in this dataset was collected in the Biocatalysis Laboratory of the Delft University of Technology - Faculty of Applied Sciences, between September 2019 and September 2024.

**Test equipment***
Test equipment information could be found in the supplementary information of the chapter.

***Description of the data in this data set***
Corresponding methods and conditions could be found in the supporting information of the chapter.

zip folders:
Ch4E01 FuncLib output.zip (this folder includes output from the computational tool FuncLib to design mutants, with corresponding input data, Excel with the best ranked mutants and pdbs of the best mutants. this was done with mutations on position 142 (serine to glutamic acid), position 155 (tyrosine to glutamic acid) or on both positions (tyrosine to phenylalanine, serine to glutamic acid))

Ch4E02 CloneManager A94D A94E LbADH alignment.zip (after doing a single mutation A94D and A94E, we confirmed the new protein sequence by determining the DNA-plasmid sequences of both mutants, the wildtype enzyme and the combination of the three)

Ch4E03 SDS-PAGEs (all gels and corresponding information are present in the Supporting Information attached in this dataset)

Ch4E04 activity assays.zip (all data was determined by UV-VIS) this folder contains the following data:
activty assays LbADH reaction buffers, activty assays in ranging reaction buffers and pH
activity assays LbADH storage conditions days incubation, activity assay in different storage conditions and activity measured of the LbADH over time 

Ch4E05 Conversions LbADH screening.zip, all data is determined by GC-FID. Folder contains the following documents:

Tyrosine mutants screening: substrate scope screened with three tyrosine mutant enzymes (see SI for context of enzymes used)
2021-02-01 results serine mutants screening: substrate scope screened with three serine mutant enzymes (see SI for context of enzymes used)
conversions A94E and A94D: substrate scope screened with single mutant enzymes (see SI for context of enzymes used)
2024-02-12 combined conversion tyrosine serine mutants LbADH: overview of all data also used in supporting information