Biosynthesis of (R)-(-)-1-Octen-3-ol in Recombinant Saccharomyces cerevisiae with Lipoxygenase-1 and Hydroperoxide Lyase Genes from Tricholoma matsutake

Abstract

IntroductionTricholoma matsutake, found in the Korean peninsula, Japan and China, have been regarded as precious food for a long time. Nowadays, it is famous symbol viand in Korea and Japan. T. matsutake is high-value food because not only its taste and smell but also its notoriously difficulty of artificial cultivation. The fruiting body of T. matsutake is formed by the complex interaction of several ecological factors in specific forest of Pinus densiflora [11,21]. Unfortunately, the occurrence of T. matsutake fruiting body in nature is steadily decreasing and the demand for the artificial cultivation of T. matsutake is rising. In 1983, Hiroshima Forestry Examination Center had succeeded in the artificial cultivation but failed to preserving this cultivating artifact. Since 1983, lots of studies have been reported and recently the National Forest Research Institute, Korea and Taki Chemical Industry, Japan had succeeded the artificial cultivation of T. matsutake in 2010 and 2018, respectively [17,20]. But most of studies had failed to maintain the cultivation and little is known about its artificial cultivation [23,31]. Moreover, transplantation of P. densiflora trees infected with T. matsutake attempted by Japan and Korea had failed to yield the mushroom with a meaningful number [12,16]. The genomic study of T. matsutake was carried out by Dr Min's research group in January 2020 [28]. Thus, in this study, not artificial cultivation but rather production of the T. matsutake flavor is the focus of this study. We expect the outcome of our work to help producing the (R)-(-)-1-octen-3-ol at a low

The Biosynthesis of 1-octene-3-ol by a Multifunctional Fatty Acid Dioxygenase and Hydroperoxide Lyase in Agaricus bisporus

“…The proteins encoded by AbLDS1 and AbLDS2 both contained DOX and CYP domains. Their CYP domains were similar to those of the PpoC of A. nidulans [ 33 ], the PpoC of A. luchuensis [ 34 ], and the LOX1 of matsutake [ 18 ] with no intact conserved heme-binding region, and were therefore nonfunctional. In contrast, the CYP domain of A. nidulans PpoA had an intact conserved heme-binding region and was functional [ 33 ] ( Figure S2 ).…”

“…A small number of identified HPLs from animals were the CYP74 clan proteins [ 53 , 54 , 55 ]. An HPL cloned from matsutake was considered to be capable of cleaving HPOD to form 1-octen-3-ol [ 18 ], which was highly homologous to sterol C-22 desaturase encoded by CYP61 in Saccharomyces cerevisiae [ 56 ]. Whether sterol C-22 desaturase has multisubstrate properties was not recorded.…”

“…Subsequently, this pathway was also found in A. luchuensis [ 34 , 35 ]. Alternatively, the LOX and HPL from T. matsutake co-expressed by yeast can catalyze the conversion of linoleic acid to produce 1-octene-3-ol [ 18 ]. However, the conserved domains in this LOX show that it is actually DOX-(CYP), and the precursor of 1-octene-3-ol was also not determined.…”

“…The 1-octen-3-ol produced by the natural oxidation of linoleic acid or chemical synthetization is a mixture of two enantiomers of (R)-(−)-1-octen-3-ol and (S)-(+)-1-octen-3-ol in equal amounts [ 18 ], while (R)-(−)-1-octen-3-ol is predominantly produced by biosynthesis, especially in button mushrooms (up to 99% of (R)-(−)-1-octen-3-ol) [ 10 ]. (S)-(+)-1-octen-3-ol has a moldy, grassy aroma [ 19 ] and low biological activity [ 20 ].…”

Chemoenzymatic Cascade Conversion of Linoleic Acid into a Secondary Fatty Alcohol Using a Combination of 13S-Lipoxygenase, Chemical Reduction, and a Photo-Activated Decarboxylase

“…The keto fatty acids could be further converted into amino fatty acids ( 4 and 5 ) by transaminases , or into medium-chain fatty acids [e.g., 9-hydroxynonanoic acid ( 6 ) and non-3­( Z )-enoic acid ( 7 ) or 12-hydroxydodec-9­( Z )-enoic acid ( 8 ) and n -hexanoic acid ( 9 )], via ester formation by Baeyer–Villiger monooxygenase (BVMO). ,,,, Linoleic acid was also converted into (6 Z ,9 Z )-heptadecadiene ( 10 ) and 5,8-dihydroxy-(9 Z ,12 Z )-octadecadienoic acid ( 11 ) by the photo-activated decarboxylase from Chlorella variabilis NC64A (Cv-FAP) − and the linoleate 5,8-diol synthases from Aspergillus nidulans, respectively (Scheme ). Moreover, linoleic acid serves as a starting material for the production of industrially relevant oxylipins such as hexanal ( 12 ) and 12-keto-9­( Z )-dodecenoic acid ( 13 ) as well as 3­( Z )-nonenal ( 14 ) and 9-oxononanoic acid ( 15 ) by serial reactions of LOXs and hydroperoxide lyases ,− (Scheme and Supporting Information, Scheme S1).…”