zymomonas mobilis硫酸盐还原途径突变菌株的构建及其产h2s与产醇评价-construction of zymomonas mobis sulfate reducing pathway mutant strain and evaluation of its h2s and alcohol production.docxVIP

ABSTRACTAssimilatory sulfate reduction pathway is one of the ways to produce H2S for microorganisms. Though this pathway in Zymomonas mobilis has been shown by results of bioinformatic research, and the relating genes have been predicted, however, research about the pathway including verification of function of the genes and regulation of the metabolic approach on experiment level has not been reported yet. At the same time, we found that Z. mobilis ZM4 could generate the irritant odour gas in the process of ethanol production from sweet sorghum juice and it proved to contain H2S as the main component. Therefore, constructing the gene-inactivated mutant strains of Z. mobilis deficient in sulfate reduction pathway and investigating their H2S production, would be of greatly scientific significance to verify the assimilatory sulfate reduction pathway and function of the relating genes, and further to recognize the whole sulfur metabolism in Z. mobilis.With a method of homologous recombination and FLP-FRT based site-specific recombination in this study, we firstly constructed a series of mutant strains of Z. mobilis deficient in the assimilatory sulfate reduction pathway. And they were ATP-sulfurylase (coded by cysN and cysD) inactivated ?cysND-cat and ?cysND, and sulfite reductase (coded by cysI and cysJ) inactivated ?cysIJ-cat and ?cysIJ, as well as the two enzyme co-inactivated ?cysND-cat?cysIJ and ?cysND?cysIJ-cat.This research tested the H2S production of all the mutant strains in different carbon source-containing medium (glucose, sucrose, fructose and sweet sorghum juice). And results showed that all the mutant strains did not produce H2S while the control ZM4 could generate H2S in all the medium except the fructose-containing medium at the condition tested. While SO32- was added, only mutant strains?cysND-cat and ?cysND could restore H2S production but it was not the case for theother mutant strains, indicating that cysND and