Isoprene is the simplest terpene and an important raw material for the rubber, food, pharmaceutical, and cosmetic industries. The construction of the mevalonate metabolic pathway in microorganisms is currently an effective way to produce isoprene and its derived terpenes, however, it can be limited by the activity and expression levels of key enzymes. Aiming at the low activity of isopentenyl pyrophosphate isomerase (IDI) and other problems, in this study, the expression of IDI was optimized from three aspects: enzyme engineering, promoter engineering and ribosome binding site (RBS) engineering. Firstly, an isoprene production system was established in Escherichia coli. IDI from Streptococcus pneumoniae was then selected for protein modeling and structural analysis, and the functional hotspot Met146 was computationally obtained and subjected to site-directed mutagenesis. The modification resulted in a mutant strain Met146His, which enhanced isoprene production to 820.46 mg/L and was 2.45 times higher than of the wild type. Next, the promoter optimization and the RBS optimization were performed for this mutant, and a final optimized strain obtained enhanced isoprene production to 862.79 mg/L, which was 2.58 times higher than that of the starting strain. The results demonstrate it is an efficient strategy to improve isoprene production by performing enzyme molecular modification and expression optimization for IDI to improve enzyme activity and regulate expression levels. The modification of IDI can also be applied to the biosynthesis of downstream products such as sesquiterpenes and diterpenes.
SUN Li
,
LIU Chunli
,
LIU Xiuxia
,
LI Ye
,
YANG Yankun
,
BAI Zhonghu
. Modification and expression optimization of isopentenyl pyrophosphate isomerase to improve isoprene production[J]. Food and Fermentation Industries, 2023
, 49(21)
: 1
-8
.
DOI: 10.13995/j.cnki.11-1802/ts.031472
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