Table 3.

Estimation Of The Impact That Individual Factors Have On The Final Activity Improvement Observed with 237Δ-TEV Protease.^a

Coding sequenceb	Protein	Lac repressor promoter	Expression Temperature	Induction Method	RILP codon adaptation	Multiplicative Fold Improvementc	Activity Initial/Finalμmol/hr/Ld
Starting Conditione	Full Length	lacIQ	25 °C	Auto	-
Alternatef	237Δ	lacI	37 °C	IPTG	+
MHT	1.7	1.3	1.6	0.4	1.3	5	36 / 169
HT	2.3	3.9	2.7	0.3	1.3	32	4.8 / 154
GT	2.1	1.5	0.5	1.5	1.4	7	6.5 / 77

^aThe improvement was quantified by assay of TEV protease activity in cell lysates.

^bCoding sequence for the TEV variant placed into the plasmid shown in Figure 2.

^cThe multiplicative fold improvement is the product of the factors of all alternate conditions that led to higher activity.

^dThe initial activity is from the full length TEV protease and the final activity is from the optimized expression conditions.

^eThe starting condition was expression of the full-length TEV protease at 25 °C using auto-induction medium, no RILP codon adaptation and lacI^q control of LacI expression.

^fThe initial change to alternate conditions was incorporation of the 237Δ truncation. Other alternate conditions leading to higher activity are shown in bold.