Monoenzyme mutant prepared through replacement of higher plant ACS double-enzyme activity key structural domain and application of monoenzyme mutant

Abstract

The invention is applicable to the technical field of gene engineering, and provides a single enzyme mutant prepared by replacing a higher plant ACS double enzyme activity key structural domain, which comprises a BOX2 conserved domain with a conserved sequence of F-Q-D-Y-H-G-[LM]-[PSKL], a BOX6 conserved domain with a conserved sequence of M-S-S-F-[GT]-L-[VI]-S-S-Q-T-Q, and a BOX4 conserved domain with a conserved sequence of T-N-P-S-N-P-L-G-[TA]-[TAMVIF]. After the conserved domain sequences are subjected to recombination replacement, the ACS recombinant protein only has ACS or C-S lyase single enzyme activity. The key structural domain influencing the ACS and C-S lyase activity of higher plant ACS protein is identified, a key target spot is provided for controlling multiple life processes in which ethylene participates by utilizing a genetic engineering method, and wide application prospects and important economic values are achieved.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to the preparation and application of a single-enzyme mutant by substituting a double-enzyme activity key domain of ACS in higher plants. Background technique [0002] Ethylene is one of the oldest plant hormones, which plays an important role in regulating many aspects of the growth and development of higher plants, including seed germination, cell elongation, root development, leaf extension, flowering and fruit ripening, organ senescence and abscission, etc. . At the same time, the role of ethylene in the response and resistance of plants to biotic and abiotic stresses is also indispensable. In seed plants, the biosynthesis of ethylene mainly undergoes three important catalytic reactions: in the first step, methionine generates S-adenosylmethionine under the action of S-adenosylmethionine synthase ( SAM); in the second step, under the catalysis of ACC sy...

AI Technical Summary

Problems solved by technology

For a long time, it has been agreed that ACS, the key enzyme of ethylene synthesis, is a single enzyme that can only catalyze the formation of ACC from SAM. There is no report on other aspects of the activity of ACS, the key enzyme of ethylene synthesis.

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