Journal
JOURNAL OF FUNGI
Volume 8, Issue 8, Pages -Publisher
MDPI
DOI: 10.3390/jof8080827
Keywords
button mushroom; 1-octen-3-ol; linoleic acid; enzymatic pathway
Categories
Funding
- Science and Technology Department of Henan Province [202102110044, 222102110302]
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In this study, the genes involved in the biosynthesis of 1-octen-3-ol in Agaricus bisporus, including AbLOX, AbLDS1, AbLDS2, and AbHPL, were identified and cloned. AbLOX and AbHPL were found to catalyze the conversion of linoleic acid to 1-octen-3-ol. Reduced AbLOX expression was correlated with a decrease in 1-octen-3-ol yield in antisense AbLOX transformants.
The biosynthetic pathway from linoleic acid to 1-octen-3-ol in Agaricus bisporus has long been established, in which linoleic acid is converted to 10-hydroperoxide (10-HPOD) by deoxygenation, and 10-HPOD is subsequently cleaved to yield 1-octene-3-ol and 10-oxodecanoic acid. However, the corresponding enzymes have not been identified and cloned. In the present study, four putative genes involved in oxylipid biosynthesis, including one lipoxygenase gene named AbLOX, two linoleate diol synthase genes named AbLDS1 and AbLDS2, and one hydroperoxide lyase gene named AbHPL were retrieved from the A. bisporus genome by a homology search and cloned and expressed prokaryotically. AbLOX, AbLDS1, and AbLDS2 all exhibited fatty acid dioxygenase activity, catalyzing the conversion of linoleic acid to generate hydroperoxide, and AbHPL showed a cleaving hydroperoxide activity, as was determined by the KI-starch method. AbLOX and AbHPL catalyzed linoleic acid to 1-octen-3-ol with an optimum temperature of 35 degrees C and an optimum pH of 7.2, whereas AbLDS1, AbLDS2, and AbHPL catalyzed linoleic acid without 1-octen-3-ol. Reduced AbLOX expression in antisense AbLOX transformants was correlated with a decrease in the yield of 1-octen-3-ol. AbLOX and AbHPL were highly homologous to the sesquiterpene synthase Cop4 of Coprinus cinerea and the yeast sterol C-22 desaturase, respectively. These results reveal that the enzymes for the oxidative cleavage of linoleic acid to synthesize 1-octen-3-ol in A. bisporus are the multifunctional fatty acid dioxygenase AbLOX and hydroperoxide lyase AbHPL.
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