目的 紫芝Ganoderma sinense次生代謝產(chǎn)物含量較低，篩選一種可以顯著提高紫芝次生代謝物產(chǎn)量的方法。方法 基于紫芝基因組數(shù)據(jù)庫篩選出編碼芳樟醇合酶的基因GsSTS50，通過改善融合酶、共表達基因和引入異源甲羥戊酸（mevalonate pathway，MVA）代謝途徑等方法使GsSTS50基因在大腸桿菌中高效表達，利用頂空固相微萃取-氣相色譜-質(zhì)譜聯(lián)用（HS-SPME-GC-MS）檢測大腸桿菌培養(yǎng)物中芳樟醇的含量。結(jié)果 優(yōu)化融合酶的策略，使大腸桿菌中芳樟醇產(chǎn)量提高了約4.9倍；通過引入異源MVA途徑，改造后菌株芳樟醇產(chǎn)量比初始培養(yǎng)提高了約49倍；將多個基因共表達的同時引入異源途徑，優(yōu)化菌株芳樟醇產(chǎn)量比原始菌株提高了42倍。結(jié)論 經(jīng)過改造的重組菌株生產(chǎn)產(chǎn)物的量均有顯著提高，為后續(xù)靈芝等擔子菌中萜類化合物以及其他萜類物質(zhì)產(chǎn)量的提升提供了參考。

Objective The low content of secondary metabolites of Ganoderma sinense has brought some difficulties to the development of monomers, so it is necessary to screen a method that can significantly increase the yield of secondary metabolites of G. sinense. Methods The GsSTS50 gene encoding linalool synthetase was selected based on the G. sinense gene library. The GsSTS50 gene was overexpressed in Escherichia coli by co-expression and introduction of heterologous mevalonate (MVA) pathway. The linalool content in E. coli was determined by HS-SPME-GC-MS. Results The yield of linalool and other terpenoids in E. coli was increased by about 4.9 times by optimizing the strategy of fusion enzyme. By introducing heterologous MVA pathway, the production of linalool was 49 times higher than that of the original culture. The co-expression of multiple genes at the same time into the heterologous pathway, the yield of the optimized strain was 42 times higher than that of the original strain. Conclusion The results showed that the yield of recombinant strains was significantly increased. This study provides a reference for further research on terpenoids and other terpenoids from G. sinense.

R286.2

國家自然科學基金資助項目（81603221）